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Identification of avaC from Human Gut Microbial Isolates that Converts 5AVA to 2-Piperidone
Qiudi Zhou, Lihui Feng
J. Microbiol. 2024;62(5):367-379.   Published online June 17, 2024
DOI: https://doi.org/10.1007/s12275-024-00141-0
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AbstractAbstract PDF
2-piperidone is a crucial industrial raw material of high-value nylon-5 and nylon-6,5. Currently, a major bottleneck in the biosynthesis of 2-piperidone is the identification of highly efficient 2-piperidone synthases. In this study, we aimed to identify specific strains among 51 human gut bacterial strains capable of producing 2-piperidone and to elucidate its synthetic mechanism. Our findings revealed that four gut bacterial strains, namely Collinsella aerofaciens LFYP39, Collinsella intestinalis LFYP54, Clostridium bolteae LFYP116, and Clostridium hathewayi LFYP18, could produce 2-piperidone from 5-aminovaleric acid (5AVA). Additionally, we observed that 2-piperidone could be synthesized from proline through cross-feeding between Clostridium difficile LFYP43 and one of the four 2-piperidone producing strains, respectively. To identify the enzyme responsible for catalyzing the conversion of 5AVA to 2-piperidone, we utilized a gain-of-function library and identified avaC (5-aminovaleric acid cyclase) in C. intestinalis LFYP54. Moreover, homologous genes of avaC were validated in the other three bacterial strains. Notably, avaC were found to be widely distributed among environmental bacteria. Overall, our research delineated the gut bacterial strains and genes involved in 2-piperidone production, holding promise for enhancing the efficiency of industrial biosynthesis of this compound.

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  • Metabolite biomarkers of screening neonatal congenital hypothyroidism based on dried blood spot metabolomics
    Xingyu Guo, Feng Suo, Yuting Wang, Di Yu, Yi Wang, Bulian Dong, Lingshan Gou, Xinhui Gan, Benjing Wang, Chaowen Yu, Xiaoxiang Xie, Dandan Linghu, Xinyu Liu, Maosheng Gu, Guowang Xu
    Analytical and Bioanalytical Chemistry.2025; 417(13): 2889.     CrossRef
  • Scarless gene disruption enabled by a dual-plasmid knockout platform in a clinical infant-derived Bifidobacterium breve strain
    Zhenxuan Gao, Lihui Feng
    Frontiers in Microbiology.2025;[Epub]     CrossRef
Repeated Exposure of Vancomycin to Vancomycin-Susceptible Staphylococcus aureus (VSSA) Parent Emerged VISA and VRSA Strains with Enhanced Virulence Potentials
An Nguyen, J Jean Sophy Roy, Ji-Hoon Kim, Kyung-Hee Yun, Wonsik Lee, Kyeong Kyu Kim, Truc Kim, Akhilesh Kumar Chaurasia
J. Microbiol. 2024;62(7):535-553.   Published online May 30, 2024
DOI: https://doi.org/10.1007/s12275-024-00139-8
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AbstractAbstract PDF
The emergence of resistance against the last-resort antibiotic vancomycin in staphylococcal infections is a serious concern for human health. Although various drug-resistant pathogens of diverse genetic backgrounds show higher virulence potential, the underlying mechanism behind this is not yet clear due to variability in their genetic dispositions. In this study, we investigated the correlation between resistance and virulence in adaptively evolved isogenic strains. The vancomycin-susceptible Staphylococcus aureus USA300 was exposed to various concentrations of vancomycin repeatedly as a mimic of the clinical regimen to obtain mutation(s)-accrued-clonally-selected (MACS) strains. The phenotypic analyses followed by expression of the representative genes responsible for virulence and resistance of MACS strains were investigated. MACS strains obtained under 2 and 8 µg/ml vancomycin, named Van2 and Van8, respectively; showed enhanced vancomycin minimal inhibitory concentrations (MIC) to 4 and 16 µg/ml, respectively. The cell adhesion and invasion of MACS strains increased in proportion to their MICs. The correlation between resistance and virulence potential was partially explained by the differential expression of genes known to be involved in both virulence and resistance in MACS strains compared to parent S. aureus USA300. Repeated treatment of vancomycin against vancomycin-susceptible S. aureus (VSSA) leads to the emergence of vancomycin-resistant strains with variable levels of enhanced virulence potentials.

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  • Targeting the G-quadruplex as a novel strategy for developing antibiotics against hypervirulent drug-resistant Staphylococcus aureus
    Maria Sultan, Maria Razzaq, Joohyun Lee, Shreyasi Das, Shrute Kannappan, Vinod Kumar Subramani, Wanki Yoo, Truc Kim, Hye-Ra Lee, Akhilesh K. Chaurasia, Kyeong Kyu Kim
    Journal of Biomedical Science.2025;[Epub]     CrossRef
  • Staphylococcus parequorum sp. nov. and Staphylococcus halotolerans sp. nov., isolated from traditional Korean soybean foods
    Ju Hye Baek, Dong Min Han, Dae Gyu Choi, Chae Yeong Moon, Jae Kyeong Lee, Chul-Hong Kim, Jung-Woong Kim, Che Ok Jeon
    Journal of Microbiology.2025; 63(8): e2503003.     CrossRef
Recombinant Protein Mimicking the Antigenic Structure of the Viral Surface Envelope Protein Reinforces Induction of an Antigen‑Specific and Virus‑Neutralizing Immune Response Against Dengue Virus
Ju Kim , Tae Young Lim , Jisang Park , Yong&#
J. Microbiol. 2023;61(1):131-143.   Published online February 1, 2023
DOI: https://doi.org/10.1007/s12275-023-00021-z
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AbstractAbstract PDF
Dengue virus (DENV), belonging to the family Flaviviridae, is the causative agent of dengue and comprises four serotypes. A second heterologous DENV infection is a critical risk factor for severe dengue, and no effective vaccine is available to prevent infection by all four DENV serotypes. Recombinant DENV vaccines are primarily based on the envelope proteins, prM and E. The E protein and its envelope domain III (EDIII) have been investigated as candidate antigens (Ags) for recombinant subunit vaccines. However, most EDIII-based Ags are monomers that do not display the cognate antigenic structure of E protein, which is essential for induction of virus-neutralizing immunity. Here, we developed recombinant DENV-2 envelope domain (r2ED) protein as an Ag that mimics the quaternary structure of E protein on the DENV surface. We confirmed that r2ED retained the conformational epitope displayed at the E-dimer interface, which reportedly exhibits broad virus-neutralizing capacity, without displaying the fusion loop epitope that causes antibody (Ab)-dependent enhancement. Furthermore, compared with EDIII alone, r2ED elicited stronger Ag-specific and cross-reactive neutralizing Ab and T cell-mediated immune responses in mice. This Ag-specific immunity was maintained at an elevated level 6 months after the last immunization, suggesting sustained Ag-specific immune memory. Taken together, these observations suggest that r2ED could be used to develop an improved subunit vaccine capable of inducing a broadly cross-reactive and long-lasting immune response against DENV infection.

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  • Antiviral effects of heme oxygenase-1 against canine coronavirus and canine influenza virus in vitro
    Jae-Hyeong Kim, Dong-Hwi Kim, Kyu-Beom Lim, Joong-Bok Lee, Seung-Yong Park, Chang-Seon Song, Sang-Won Lee, Dong-Hun Lee, Do-Geun Kim, Hun-Young Yoon, In-Soo Choi
    Journal of Microbiology.2025; 63(5): e2501029.     CrossRef
  • A retrospective computational validation of a clinically evaluated recombinant envelope protein tetravalent dengue vaccine
    Jesús Reiné, Rosaria Tinnirello, Alberto Cagigi, Chiuan Yee Leow, Chiuan Herng Leow, Gioacchin Iannolo, Bruno Douradinha
    International Journal of Biological Macromolecules.2025; 329: 147688.     CrossRef
  • Peptides of a Feather: How Computation Is Taking Peptide Therapeutics under Its Wing
    Thomas David Daniel Kazmirchuk, Calvin Bradbury-Jost, Taylor Ann Withey, Tadesse Gessese, Taha Azad, Bahram Samanfar, Frank Dehne, Ashkan Golshani
    Genes.2023; 14(6): 1194.     CrossRef
Characterization and bioefficacy of green nanosilver particles derived from fungicide-tolerant Tricho-fusant for efficient biocontrol of stem rot (Sclerotium rolfsii Sacc.) in groundnut (Arachis hypogaea L.)
Darshna G. Hirpara , Harsukh P. Gajera , Disha D. Savaliya , Rushita V. Bhadani
J. Microbiol. 2021;59(11):1031-1043.   Published online October 6, 2021
DOI: https://doi.org/10.1007/s12275-021-1344-9
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AbstractAbstract PDF
An efficient and eco-friendly bioefficacy of potent Trichofusant (Fu21) and its green nanosilver formulation against stem rot (Sclerotium rolfsii) in groundnut was established. Fu21 demonstrated higher in-vitro growth inhibition of pathogen with better fungicide tolerance than the parental strains. The green nanosilver particles were synthesized from the extracellular metabolites of Fu21 and characterized for shape (spherical, 59.34 nm in scanning electron microscope), purity (3.00 KeV, energy dispersive X-ray analysis), size (54.3 nm in particle size analyzer), and stability (53.7 mv, zeta). The field efficacy study exhibited that the seedling emergence was high in seeds treated with green nanosilver (minimum inhibitory concentration-[MIC] 20 μg Ag/ml), and a low disease severity index of stem rot during the crop growth was followed by the live antagonist (Fu21) in addition to seed treatment with a fungicide mix under pathogen infestation. The seed quality analysis of harvested pods revealed a high oil content with balanced fatty acid composition (3.10 oleic/linoleic acid ratio) in green nanosilver treatment under pathogen infestation. The residual analysis suggested that green nanosilver applied at the MIC level as seed treatment yielded similar effects as the control for silver residue in the harvested groundnut seeds. The green nanosilver at MIC has a high pod-yield under S. rolfsii infestation, demonstrating green chemistry and sustainability of the nanoproduct.

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  • Comparative impact of seed priming with zinc oxide nanoparticles and zinc sulphate on biocompatibility, zinc uptake, germination, seedling vitality, and antioxidant modulation in groundnut
    M. N. Ashwini, H. P. Gajera, Darshna G. Hirpara, Disha D. Savaliya, U. K. Kandoliya
    Journal of Nanoparticle Research.2024;[Epub]     CrossRef
  • Biotechnological trends and optimization of Arachis hypogaea residues valorization: A bibliometric analysis and comprehensive review
    Oyetola Ogunkunle, Micheal Olusoji Olusanya
    Bioresource Technology.2024; 414: 131585.     CrossRef
  • Intracellular metabolomics and microRNAomics unveil new insight into the regulatory network for potential biocontrol mechanism of stress‐tolerant Tricho‐fusants interacting with phytopathogen Sclerotium rolfsii Sacc
    Darshna G. Hirpara, Harsukh P. Gajera
    Journal of Cellular Physiology.2023; 238(6): 1288.     CrossRef
  • Antifungal Properties of Nanosilver Clay Composites Against Fungal Pathogens of Agaricus bisporus
    Tebogo Levy Ramakutoane, Magaretha Petronella Roux‐van der Merwe, Jacqueline Badenhorst, Sreejarani Kesavan Pillai, Suprakas Sinha Ray
    ChemistrySelect.2023;[Epub]     CrossRef
  • Exploring conserved and novel MicroRNA-like small RNAs from stress tolerant Trichoderma fusants and parental strains during interaction with fungal phytopathogen Sclerotium rolfsii Sacc.
    Darshna G. Hirpara, H.P. Gajera, Disha D. Savaliya, M.V. Parakhia
    Pesticide Biochemistry and Physiology.2023; 191: 105368.     CrossRef
  • Biochemical and molecular depictions to develop ech42 gene-specific SCAR markers for recognition of chitinolytic Trichoderma inhibiting Macrophomina phaseolina (Maubl.) Ashby
    H. P. Gajera, Darshna G. Hirpara, Disha D. Savaliya, M. V. Parakhia
    Archives of Microbiology.2023;[Epub]     CrossRef
  • Salicylic acid-functionalised chitosan nanoparticles restore impaired sucrose metabolism in the developing anther of cotton (Gossypium hirsutum) under heat stress
    Khyati R. Savani, H. P. Gajera, Darshna G. Hirpara, Disha D. Savaliya, U. K. Kandoliya, Honghong Wu
    Functional Plant Biology.2023; 50(9): 736.     CrossRef
Incomplete autophagy promotes the replication of Mycoplasma hyopneumoniae
Zhaodi Wang† , Yukang Wen† , Bingqian Zhou , Yaqin Tian , Yaru Ning , Honglei Ding
J. Microbiol. 2021;59(8):782-792.   Published online July 5, 2021
DOI: https://doi.org/10.1007/s12275-021-1232-3
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AbstractAbstract PDF
Autophagy is an important cellular homeostatic mechanism for recycling of degradative proteins and damaged organelles. Autophagy has been shown to play an important role in cellular responses to bacteria and bacterial replication. However, the role of autophagy in Mycoplasma hyopneumoniae infection and the pathogenic mechanism is not well characterized. In this study, we showed that M. hyopneumoniae infection significantly increases the number of autophagic vacuoles in host cells. Further, we found significantly enhanced expressions of autophagy marker proteins (LC3-II, ATG5, and Beclin 1) in M. hyopneumoniae-infected cells. Moreover, immunofluorescence analysis showed colocalization of P97 protein with LC3 during M. hyopneumoniae infection. Interestingly, autophagic flux marker, p62, accumulated with the induction of infection. Conversely, the levels of p62 and LC3-II were decreased after treatment with 3-MA, inhibiting the formation of autophagosomes, during infection. In addition, accumulation of autophagosomes promoted the expression of P97 protein and the survival of M. hyopneumoniae in PK- 15 cells, as the replication of M. hyopneumoniae was downregulated by adding 3-MA. Collectively, these findings provide strong evidence that M. hyopneumoniae induces incomplete autophagy, which in turn enhances its reproduction in host cells. These findings provide novel insights into the interaction of M. hyopneumoniae and host.

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  • Proteomics Reveals the Response Mechanism of Embryonic Bovine Lung Cells to Mycoplasma bovis Infection
    Li Wang, Qing Wang, Yudong Liu, Yunxia Chen, Shijun Bao, Xiaoli Zhang, Chuan Wang
    International Journal of Molecular Sciences.2025; 26(2): 823.     CrossRef
  • Omega-3 PUFAs improve cognitive function in heat-stressed mice by enhancing autophagy via inhibition of the phosphorylation of the PI3K–Akt–mTOR pathway
    Zifu Ren, Mengyu Cai, Xinyao Liu, Xin Li, Wenjing Shi, Hongtao Lu, Hui Shen, Gen Miao, Qicheng Zhou, Hongxia Li
    Food & Function.2025; 16(5): 1931.     CrossRef
  • The DHH-DHHA1 domain phosphodiesterase of Mycoplasma bovis employs multiple strategies to modulate macrophage cellular processes
    Xifang Zhu, Ye Chen, Doukun Lu, Gang Zhao, Yankai Liu, Aiping Wang, Aizhen Guo
    International Journal of Biological Macromolecules.2025; 306: 141585.     CrossRef
  • Research Progress on Immune Evasion of Mycoplasma hyopneumoniae
    Bin Jiang, Ying Zhang, Gaojian Li, Yanping Quan, Jianhong Shu, Huapeng Feng, Yulong He
    Microorganisms.2024; 12(7): 1439.     CrossRef
  • The Role of Pyroptosis and Autophagy in Ischemia Reperfusion Injury
    Huijie Zhao, Yihan Yang, Xinya Si, Huiyang Liu, Honggang Wang
    Biomolecules.2022; 12(7): 1010.     CrossRef
  • Mycoplasma bovis inhibits autophagy in bovine mammary epithelial cells via a PTEN/PI3K-Akt-mTOR-dependent pathway
    Maolin Xu, Yang Liu, Tuerdi Mayinuer, Yushan Lin, Yue Wang, Jian Gao, Dong Wang, John P. Kastelic, Bo Han
    Frontiers in Microbiology.2022;[Epub]     CrossRef
  • Incomplete autophagy promotes the proliferation of Mycoplasma hyopneumoniae through the JNK and Akt pathways in porcine alveolar macrophages
    Yukang Wen, Zhengkun Chen, Yaqin Tian, Mei Yang, Qingshuang Dong, Yujiao Yang, Honglei Ding
    Veterinary Research.2022;[Epub]     CrossRef
Different distribution patterns of microorganisms between aquaculture pond sediment and water
Lili Dai , Chengqing Liu , Liang Peng , Chaofeng Song , Xiaoli Li , Ling Tao
J. Microbiol. 2021;59(4):376-388.   Published online February 25, 2021
DOI: https://doi.org/10.1007/s12275-021-0635-5
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AbstractAbstract PDF
Aquatic microorganisms in the sediment and water column are closely related; however, their distribution patterns between these two habitats still remain largely unknown. In this study, we compared sediment and water microeukaryotic and bacterial microorganisms in aquaculture ponds from different areas in China, and analyzed the influencing environmental factors as well as the inter-taxa relationships. We found that bacteria were significantly more abundant than fungi in both sediment and water, and the bacterial richness and diversity in sediment were higher than in water in all the sampling areas, but no significant differences were found between the two habitats for microeukaryotes. Bacterial taxa could be clearly separated through cluster analysis between the sediment and water, while eukaryotic taxa at all classification levels could not. Spirochaetea, Deltaproteobacteria, Nitrospirae, Ignavibacteriae, Firmicutes, Chloroflexi, and Lentimicrobiaceae were more abundantly distributed in sediment, while Betaproteobacteria, Alphaproteobacter, Cyanobacteria, Roseiflexaceae, Dinghuibacter, Cryomorphaceae, and Actinobacteria were more abundant in water samples. For eukaryotes, only Cryptomonadales were found to be distributed differently between the two habitats. Microorganisms in sediment were mainly correlated with enzymes related to organic matter decomposition, while water temperature, pH, dissolved oxygen, and nutrient levels all showed significant correlation with the microbial communities in pond water. Intensive interspecific relationships were also found among eukaryotes and bacteria. Together, our results indicated that eukaryotic microorganisms are distributed less differently between sediment and water in aquaculture ponds compared to bacteria. This study provides valuable data for evaluating microbial distributions in aquatic environments, which may also be of practical use in aquaculture pond management.

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  • Heterogeneous bacterial communities in gills and intestine of Nile tilapia (Oreochromis niloticus) and in water and sediments of aquaculture ponds in Bangladesh
    Anwar Hossain, Md Al Zahid, Santonu Kumar Sanyal, Md Inja-Mamun Haque, Md Habibullah-Al-Mamun, Shankar Chandra Mandal, Kozo Watanabe
    Aquaculture and Fisheries.2025; 10(4): 696.     CrossRef
  • Bacterial Community Composition and Prevalence of Aeromonas dhakensis in Four Tilapia Freshwater Aquaculture Systems in Malaysia
    Sook Ling Lim, Suat Moi Puah, Siti Nursyuhada Baharudin, Nur Insyirah Mohd Razalan, Kieng Soon Hii, Wei Ching Khor, Yen Ching Lim, Kyaw Thu Aung, Kek Heng Chua, Po Teen Lim, Chui Pin Leaw
    Fishes.2025; 10(5): 204.     CrossRef
  • Dynamics of soil properties and pathogen levels in Pacific white shrimp ponds during a production cycle: Implications for aquaculture management
    Suwanit Chainark, Vanida Sumetlux, Pitchaya Chainark
    Journal of the World Aquaculture Society.2025;[Epub]     CrossRef
  • Deciphering the temporal and dose-responsive dynamics of microbial communities in aquaculture mesocosms under florfenicol treatment
    Xiaoxi Deng, Chengqian Liang, Yu Wang, Yanni Geng, Jian Han, Ke Yu
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  • Efficacy of the ‘three ponds and two dams’ ecological treatment system in purifying shrimp pond wastewater and mitigating antibiotic resistance
    Bin Wei, Jie Fang, Dongxian Zhou, Xuelin Gu, Bingyao Sun, Xuehong Song
    Science of The Total Environment.2025; 992: 179928.     CrossRef
  • Positive relationship between Clostridium abundance and bacterial community diversity in the aquaculture water of hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) in coastal areas of Hainan Island
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    Israeli Journal of Aquaculture - Bamidgeh.2025;[Epub]     CrossRef
  • Intestinal microbiota responses to environmental microbiomes and factors across populations of the Manila clam Ruditapes philippinarum
    Longzhen Liu, Lulei Liu, Haonan Zhuang, Ang Li, Zirong Liu, Minghui Jiao, Jiamin Li, Suyan Xue, Jiaqi Li, Yushui Ren, Yuze Mao
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  • Bacterial community characterization of water and sediment in different culture systems of prawn Macrobrachium rosenbergii
    Meng Ni, Songbao Zou, Mei Liu, Dan Zhou, Julin Yuan
    Israeli Journal of Aquaculture - Bamidgeh.2025;[Epub]     CrossRef
  • Bacterial hazards in urban stream irrigation in peri-urban interface of Nairobi-Machakos counties, Kenya
    Arcadius Martinien Agassin Ahogle, Nicholas K. Korir, Pascal Houngnandan, Lina Abu-Ghunmi, Sammy Letema
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  • Metagenomic Insight into the Effect of Probiotics on Nitrogen Cycle in the Coilia nasus Aquaculture Pond Water
    Qi Mang, Jun Gao, Quanjie Li, Yi Sun, Gangchun Xu, Pao Xu
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  • Carbon and Nutrient Limitations of Microbial Metabolism in Xingkai Lake, China: Abiotic and Biotic Drivers
    Xingting Chen, Weizhen Zhang, Mengdie Geng, Ji Shen, Jianjun Wang
    Microbial Ecology.2024;[Epub]     CrossRef
  • Biofilm formation and chlorine resistance of microbial communities in household drinking water system: Preliminary idea of using bacteria to control bacteria
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  • Effects of Coal Mining Activities on the Changes in Microbial Community and Geochemical Characteristics in Different Functional Zones of a Deep Underground Coal Mine
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  • Effect of Pond-Based Rice Floating Bed on the Microbial Community Structure and Quality of Water in Pond of Mandarin Fish Fed Using Artificial Diet
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    Minhai Liu, Zhongneng Yan, Chengwei Huang, Zhihua Lin, Zhilan Peng, Chenxi Zhao, Xiafei Zheng
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    Antonia Bruno, Alessandra Cafiso, Anna Sandionigi, Andrea Galimberti, Davide Magnani, Amedeo Manfrin, Giulio Petroni, Maurizio Casiraghi, Chiara Bazzocchi
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    Zebing Zhu, Siyang Xu, Xiajun Bao, Lili Shan, Yunyan Pei, Wanjun Zheng, Yixing Yuan
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  • The Responses of Sediment Bacterial Communities in Chinese Mitten Crab (Eriocheir sinensis) Culture Ponds to Changes in Physicochemical Properties Caused by Sediment Improvement
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  • Antibiotic resistance gene-free probiont administration to tilapia for growth performance and Streptococcus agalactiae resistance
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  • Analysis of the structure and function of microbial community in late-stage of grass carp (Ctenopharyngodon idella) farming ponds
    Shuhui Niu, Kai Zhang, Zhifei Li, Jun Xie, Guangjun Wang, Hongyan Li, Ermeng Yu, Yun Xia, Jingjing Tian, Wangbao Gong
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  • Structural and functional comparisons of the environmental microbiota of pond and tank environments at different locations for the commercial aquaculture of American shad
    J. Du, Q.H. Liu, J.S. Liu, K.-Q. Zhang, W.F. Huang
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  • The responses of soil bacterial and archaeal communities to coastal embankments in three typical salt marshes of Eastern China
    Hongyu Feng, Yajun Qiao, Lu Xia, Wen Yang, Yongqiang Zhao, Nasreen Jeelani, Shuqing An
    Plant and Soil.2022; 477(1-2): 439.     CrossRef
  • Improvement of fish production and water quality in a recirculating aquaculture pond enhanced with bacteria-microalgae association
    Chu Wang, Cancan Jiang, Tianming Gao, Xiawei Peng, Shuanglong Ma, Qian Sun, Bing Xia, Xiangming Xie, Zhihui Bai, Shengjun Xu, Xuliang Zhuang
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  • Microbial Community Structure and Its Driving Environmental Factors in Black Carp (Mylopharyngodon piceus) Aquaculture Pond
    Xuemei Li, Lu Liu, Yongjiu Zhu, Tingbing Zhu, Xingbing Wu, Deguo Yang
    Water.2021; 13(21): 3089.     CrossRef
Role of melatonin in murine “restraint stress”-induced dysfunction of colonic microbiota
Rutao Lin , Zixu Wang , Jing Cao , Ting Gao , Yulan Dong , Yaoxing Chen
J. Microbiol. 2021;59(5):500-512.   Published online February 25, 2021
DOI: https://doi.org/10.1007/s12275-021-0305-7
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AbstractAbstract PDF
Intestinal diseases caused by physiological stress have become a severe public health threat worldwide. Disturbances in the gut microbiota-host relationship have been associated with irritable bowel disease (IBD), while melatonin (MT) has antiinflammatory and antioxidant effects. The objective of this study was to investigate the mechanisms by which MT-mediated protection mitigated stress-induced intestinal microbiota dysbiosis and inflammation. We successfully established a murine restraint stress model with and without MT supplementation. Mice subjected to restraint stress had significantly elevated corticosterone (CORT) levels, decreased MT levels in their plasma, elevated colonic ROS levels and increased bacterial abundance, including Bacteroides and Tyzzerella, in their colon tract, which led to elevated expression of Toll-like receptor (TLR) 2/4, p-P65 and p-IκB. In contrast, supplementation with 20 mg/kg MT reversed the elevation of the plasma CORT levels, downregulated the colon ROS levels and inhibited the changes in the intestinal microbiota induced by restraint stress. These effects, in turn, inhibited the activities of TLR2 and TLR4, p-P65 and p-IκB, and decreased the inflammatory reaction induced by restraint stress. Our results suggested that MT may mitigate “restraint stress”-induced colonic microbiota dysbiosis and intestinal inflammation by inhibiting the activation of the NF-κB pathway.

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Review
[MINIREVIEW]Regulation of gene expression by protein lysine acetylation in Salmonella
Hyojeong Koo , Shinae Park , Min-Kyu Kwak , Jung-Shin Lee
J. Microbiol. 2020;58(12):979-987.   Published online November 17, 2020
DOI: https://doi.org/10.1007/s12275-020-0483-8
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AbstractAbstract PDF
Protein lysine acetylation influences many physiological functions, such as gene regulation, metabolism, and disease in eukaryotes. Although little is known about the role of lysine acetylation in bacteria, several reports have proposed its importance in various cellular processes. Here, we discussed the function of the protein lysine acetylation and the post-translational modifications (PTMs) of histone-like proteins in bacteria focusing on Salmonella pathogenicity. The protein lysine residue in Salmonella is acetylated by the Pat-mediated enzymatic pathway or by the acetyl phosphate-mediated non-enzymatic pathway. In Salmonella, the acetylation of lysine 102 and lysine 201 on PhoP inhibits its protein activity and DNAbinding, respectively. Lysine acetylation of the transcriptional regulator, HilD, also inhibits pathogenic gene expression. Moreover, it has been reported that the protein acetylation patterns significantly differ in the drug-resistant and -sensitive Salmonella strains. In addition, nucleoid-associated proteins such as histone-like nucleoid structuring protein (H-NS) are critical for the gene silencing in bacteria, and PTMs in H-NS also affect the gene expression. In this review, we suggest that protein lysine acetylation and the post-translational modifications of H-NS are important factors in understanding the regulation of gene expression responsible for pathogenicity in Salmonella.

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Journal Articles
Phosphorylation of tegument protein pp28 contributes to trafficking to the assembly compartment in human cytomegalovirus infection
Jun-Young Seo , Jin Ah Heo , William J. Britt
J. Microbiol. 2020;58(7):624-631.   Published online June 27, 2020
DOI: https://doi.org/10.1007/s12275-020-0263-5
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AbstractAbstract PDF
Human cytomegalovirus (HCMV) UL99 encodes a late tegument protein pp28 that is essential for envelopment and production of infectious virus. This protein is localized to the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) in transfected cells but it localizes to the cytoplasmic assembly compartment (AC) in HCMV-infected cells. Trafficking of pp28 to the AC is required for the assembly of infectious virus. The N-terminal domain (aa 1-61) of pp28 is sufficient for trafficking and function of the wild type protein during viral infection. However, residues required for authentic pp28 trafficking with the exception of the acidic cluster in the N-terminal domain of pp28 remain undefined. Monitoring protein migration on SDS-PAGE, we found that pp28 is phosphorylated in the virus-infected cells and dephosphorylated in the viral particles. By generating substitution mutants of pp28, we showed that three serine residues (aa 41–43) and a tyrosine residue (aa 34) account for its phosphorylation. The mutant forms of pp28 were localized to the plasma membrane as well as the ERGIC in transfected cells. Likewise, these mutant proteins were localized to the plasma membrane as well as the AC in virus-infected cells. These results suggested that phosphorylation of pp28 contributes to its intracellular trafficking and efficient viral assembly and incorporation.

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Light affects picocyanobacterial grazing and growth response of the mixotrophic flagellate Poterioochromonas malhamensis
Thomas Weisse , Michael Moser
J. Microbiol. 2020;58(4):268-278.   Published online January 28, 2020
DOI: https://doi.org/10.1007/s12275-020-9567-8
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AbstractAbstract PDF
We measured the grazing and growth response of the mixotrophic chrysomonad flagellate Poterioochromonas malhamensis on four closely related picocyanobacterial strains isolated from subalpine lakes in central Europe. The picocyanobacteria represented different pigment types (phycoerythrin- rich, PE, and phycocyanin-rich, PC) and phylogenetic clusters. The grazing experiments were conducted with laboratory cultures acclimated to 10 μmol photon/m2/sec (low light, LL) and 100 μmol photon/m2/sec (moderate light, ML), either in the dark or at four different irradiances ranging from low (6 μmol photon/m2/sec) to high (1,500 μmol photon/m2/ sec) light intensity. Poterioochromonas malhamensis preferred the larger, green PC-rich picocyanobacteria to the smaller, red PE-rich picocyanobacterial, and heterotrophic bacteria. The feeding and growth rates of P. malhamensis were sensitive to the actual light conditions during the experiments; the flagellate performed relatively better in the dark and at LL conditions than at high light intensity. In summary, our
results
found strain-specific ingestion and growth rates of the flagellate; an effect of the preculturing conditions, and, unexpectedly, a direct adverse effect of high light levels. We conclude that this flagellate may avoid exposure to high surface light intensities commonly encountered in temperate lakes during the summer.

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    Thomas Weisse
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Overexpression and characterization of a novel cold-adapted and salt-tolerant GH1 β-glucosidase from the marine bacterium Alteromonas sp. L82
Jingjing Sun , Wei Wang , Congyu Yao , Fangqun Dai , Xiangjie Zhu , Junzhong Liu , Jianhua Hao
J. Microbiol. 2018;56(9):656-664.   Published online August 23, 2018
DOI: https://doi.org/10.1007/s12275-018-8018-2
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AbstractAbstract PDF
A novel gene (bgl) encoding a cold-adapted β-glucosidase was cloned from the marine bacterium Alteromonas sp. L82. Based on sequence analysis and its putative catalytic conserved region, Bgl belonged to the glycoside hydrolase family 1. Bgl was overexpressed in E. coli and purified by Ni2+ affinity chromatography. The purified recombinant β- glucosidase showed maximum activity at temperatures between 25°C to 45°C and over the pH range 6 to 8. The enzyme lost activity quickly after incubation at 40°C. Therefore, recombinant β-glucosidase appears to be a cold-adapted enzyme. The addition of reducing agent doubled its activity and 2 M NaCl did not influence its activity. Recombinant β-glucosidase was also tolerant of 700 mM glucose and some organic solvents. Bgl had a Km of 0.55 mM, a Vmax of 83.6 U/mg, a kcat of 74.3 s-1 and kcat/Km of 135.1 at 40°C, pH 7 with 4-nitrophenyl-β-D-glucopyranoside as a substrate. These properties indicate Bgl may be an interesting candidate for biotechnological and industrial applications.

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The inability of Bacillus licheniformis perR mutant to grow is mainly due to the lack of PerR-mediated fur repression
Jung-Hoon Kim , Yoon-Mo Yang , Chang-Jun Ji , Su-Hyun Ryu , Young-Bin Won , Shin-Yeong Ju , Yumi Kwon , Yeh-Eun Lee , Hwan Youn , Jin-Won Lee
J. Microbiol. 2017;55(6):457-463.   Published online April 22, 2017
DOI: https://doi.org/10.1007/s12275-017-7051-x
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AbstractAbstract PDF
PerR, a member of Fur family protein, is a metal-dependent H2O2 sensing transcription factor that regulates genes in-volved in peroxide stress response. Industrially important bac-terium Bacillus licheniformis contains three PerR-like pro-teins (PerRBL, PerR2, and PerR3) compared to its close rela-tive Bacillus subtilis. Interestingly, unlike other bacteria in-cluding B. subtilis, no authentic perRBL null mutant could be established for B. licheniformis. Thus, we constructed a con-ditional perRBL mutant using a xylose-inducible promoter, and investigated the genes under the control of PerRBL. PerRBL regulon genes include katA, mrgA, ahpC, pfeT, hemA, fur, and perR as observed for PerRBS. However, there is some variation in the expression levels of fur and hemA genes be-tween B. subtilis and B. licheniformis in the derepressed state. Furthermore, katA, mrgA, and ahpC are strongly induced, whereas the others are only weakly or not induced by H2O2 treatment. In contrast to the B. subtilis perR null mutant which frequently gives rise to large colony phenotype mainly due to the loss of katA, the suppressors of B. licheniformis perR mutant, which can form colonies on LB agar, were all cata-lase-positive. Instead, many of the suppressors showed in-creased levels of siderophore production, suggesting that the suppressor mutation is linked to the fur gene. Consistent with this, perR fur double mutant could grow on LB agar without Fe supplementation, whereas perR katA double mutant could only grow on LB agar with Fe supplementation. Taken toge-ther, our data suggest that in B. licheniformis, despite the si-milarity in PerRBL and PerRBS regulon genes, perR is an essen-tial gene required for growth and that the inability of perR null mutant to grow is mainly due to elevated expression of Fur.

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Research Support, Non-U.S. Gov'ts
Molecular characterization of mammalian-adapted Korean-type avian H9N2 virus and evaluation of its virulence in mice
Kuk Jin Park , Min-Suk Song , Eun-Ha Kim , Hyeok-il Kwon , Yun Hee Baek , Eun-hye Choi , Su-Jin Park , Se Mi Kim , Young-il Kim , Won-Suk Choi , Dae-Won Yoo , Chul-Joong Kim , Young Ki Choi
J. Microbiol. 2015;53(8):570-577.   Published online July 31, 2015
DOI: https://doi.org/10.1007/s12275-015-5329-4
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AbstractAbstract
Avian influenza A virus (AIV) is commonly isolated from domestic poultry and wild migratory birds, and the H9N2 subtype is the most prevalent and the major cause of severe disease in poultry in Korea. In addition to the veterinary concerns regarding the H9N2 subtype, it is also considered to be the next potential human pandemic strain due to its rapid evolution and interspecies transmission. In this study, we utilize serial lung-to-lung passage of a low pathogenic avian influenza virus (LPAI) H9N2 (A/Ck/Korea/163/04, WT163) (Y439-lineage) in mice to increase pathogenicity and investigate the potential virulence marker. Mouse-adapted H9N2 virus obtained high virulence (100% mortality) in mice after 98 serial passages. Sequence results show that the mouse adaptation (ma163) possesses several mutations within seven gene segments (PB2, PA, HA, NP, NA, M, and NS) relative to the wild-type strain. The HA gene showed the most mutations (at least 11) with one resulting in the loss of an N-glycosylation site (at amino acid 166). Moreover, reverse genetic studies established that an E627K substitution in PB2 and the loss of the N-glycosylation site in the HA protein (aa166) are critical virulence markers in the mouse-adapted H9N2 virus. Thus, these results add to the increasing body of mutational analysis data defining the function of the viral polymerase and HA genes and their roles in mammalian host adaptation. To our knowledge, this is first report of the generation of a mammalian-adapted Korea H9N2 virus (Y493-lineages). Therefore, this study offers valuable insights into the molecular evolution of the LPAI Korean H9N2 in a new host and adds to the current knowledge of the molecular markers associated with increased virulence.

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From the traditional Chinese medicine plant Schisandra chinensis new scaffolds effective on HIV-1 reverse transcriptase resistant to non-nucleoside inhibitors
Lijia Xu , Nicole Grandi , Claudia Del Vecchio , Daniela Mandas , Angela Corona , Dario Piano , Francesca Esposito , Cristina Parolin , Enzo Tramontano
J. Microbiol. 2015;53(4):288-293.   Published online March 4, 2015
DOI: https://doi.org/10.1007/s12275-015-4652-0
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AbstractAbstract PDF
HIV-1 reverse transcriptase (RT) is still an extremely attractive pharmaceutical target for the identification of new inhibitors possibly active on drug resistant strains. Medicinal plants are a rich source of chemical diversity and can be used to identify novel scaffolds to be further developed by chemical modifications. We investigated the ability of the main lignans from Schisandra chinensis (Turcz.) Baill. fruits, commonly used in Traditional Chinese Medicine, to affect HIV-1 RT functions. We purified 6 lignans from Schisandra chinensis fruits and assayed their effects on HIV-1 RT and viral replication. Among the S. chinensis fruit lignans, Schisandrin B and Deoxyschizandrin selectively inhibited the HIV-1 RTassociated DNA polymerase activity. Structure activity relationship revealed the importance of cyclooctadiene ring substituents for efficacy. In addition, Schisandrin B was also able to impair HIV-1 RT drug resistant mutants and the early phases of viral replication. We identified Schisandrin B and Deoxyschizandrin as new scaffold for the further development of novel HIV-1 RT inhibitors.

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    Molecular BioSystems.2017; 13(8): 1469.     CrossRef
  • Sennoside A, derived from the traditional chinese medicine plant Rheum L., is a new dual HIV-1 inhibitor effective on HIV-1 replication
    Francesca Esposito, Ilaria Carli, Claudia Del Vecchio, Lijia Xu, Angela Corona, Nicole Grandi, Dario Piano, Elias Maccioni, Simona Distinto, Cristina Parolin, Enzo Tramontano
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    Cristina Tintori, Angela Corona, Francesca Esposito, Annalaura Brai, Nicole Grandi, Elisa Rita Ceresola, Massimo Clementi, Filippo Canducci, Enzo Tramontano, Maurizio Botta
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    Maria Barbara Pisano, Sofia Cosentino, Silvia Viale, Delia Spanò, Angela Corona, Francesca Esposito, Enzo Tramontano, Paola Montoro, Carlo Ignazio Giovanni Tuberoso, Rosaria Medda, Francesca Pintus
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    Zhang qing-tian, Fan shu-tian, Yang yi-ming, Xu pei-lei, Ai jun
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    Francesca Esposito, Cristina Tintori, Riccardo Martini, Frauke Christ, Zeger Debyser, Roberto Ferrarese, Gianluigi Cabiddu, Angela Corona, Elisa Rita Ceresola, Andrea Calcaterra, Valentina Iovine, Bruno Botta, Massimo Clementi, Filippo Canducci, Maurizio
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DBA/2 Mouse as an Animal Model for Anti-influenza Drug Efficacy Evaluation
Jin Il Kim , Sehee Park , Sangmoo Lee , Ilseob Lee , Jun Heo , Min-Woong Hwang , Joon-Yong Bae , Donghwan Kim , Seok-Il Jang , Mee Sook Park , Man-Seong Park
J. Microbiol. 2013;51(6):866-871.   Published online December 19, 2013
DOI: https://doi.org/10.1007/s12275-013-3428-7
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AbstractAbstract PDF
Influenza viruses are seasonally recurring human pathogens. Vaccines and antiviral drugs are available for influenza. However, the viruses, which often change themselves via antigenic drift and shift, demand constant efforts to update vaccine antigens every year and develop new agents with broad-spectrum antiviral efficacy. An animal model is critical for such efforts. While most human influenza viruses are unable to kill BALB/c mice, some strains have been shown to kill DBA/2 mice without prior adaptation. Therefore, in this study, we explored the feasibility of employing DBA/2 mice as a model in the development of anti-influenza drugs. Unlike the BALB/c strain, DBA/2 mice were highly susceptible and could be killed with a relatively low titer (50% DBA/2 lethal dose = 102.83 plaque-forming units) of the A/ Korea/01/2009 virus (2009 pandemic H1N1 virus). When treated with a neuraminidase inhibitor, oseltamivir phosphate, infected DBA/2 mice survived until 14 days postinfection. The reduced morbidity of the infected DBA/2 mice was also consistent with the oseltamivir treatment. Taking these data into consideration, we propose that the DBA/2 mouse is an excellent animal model to evaluate antiviral efficacy against influenza infection and can be further utilized for combination therapies or bioactivity models of existing and newly developed anti-influenza drugs.

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  • DNA co-delivery of seasonal H1 influenza hemagglutinin nanoparticle vaccines with chemokine adjuvant CTACK induces potent immunogenicity for heterologous protection in vivo
    Kevin Liaw, Kylie M. Konrath, Abigail R. Trachtman, Nicholas J. Tursi, Ebony N. Gary, Cory Livingston, Kaitlyn Flowers, Jacqueline D. Chu, Casey E. Hojecki, Niklas Laenger, Madison E. McCanna, Colby J. Agostino, Neethu Chokkalingam, Kelly Bayruns, Sinja K
    Vaccine.2025; 59: 127231.     CrossRef
  • Swine influenza A virus isolates containing the pandemic H1N1 origin matrix gene elicit greater disease in the murine model
    Shelly J. Curran, Emily F. Griffin, Lucas M. Ferreri, Constantinos S. Kyriakis, Elizabeth W. Howerth, Daniel R. Perez, S. Mark Tompkins, Robert Paul de Vries
    Microbiology Spectrum.2024;[Epub]     CrossRef
  • Kinetic of the Antibody Response Following AddaVax-Adjuvanted Immunization with Recombinant Influenza Antigens
    Ted. M. Ross, Naveen Gokanapudi, Pan Ge, Hua Shi, Robert A. Richardson, Spencer R. Pierce, Pedro Sanchez, Subhan Ullah, Eliana De Luca, Giuseppe A. Sautto
    Vaccines.2022; 10(8): 1315.     CrossRef
  • Peptidylarginine Deiminase 2 in Murine Antiviral and Autoimmune Antibody Responses
    Aisha M. Mergaert, Michael F. Denny, Brock Kingstad-Bakke, Mandar Bawadekar, S. Janna Bashar, Thomas F. Warner, Marulasiddappa Suresh, Miriam A. Shelef, Baohui Xu
    Journal of Immunology Research.2022; 2022: 1.     CrossRef
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    Susana Barbosa-Méndez, Maura Matus-Ortega, Ricardo Hernández-Miramontes, Alberto Salazar-Juárez
    International Immunopharmacology.2021; 98: 107887.     CrossRef
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    Mee Sook Park, Jin Il Kim, Joon-Yong Bae, Man-Seong Park
    Laboratory Animal Research.2020;[Epub]     CrossRef
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    Gautham K Rao, Rodney A Prell, Steven T Laing, Stefanie C M Burleson, Allen Nguyen, Jacqueline M McBride, Crystal Zhang, Daniel Sheinson, Wendy G Halpern
    Toxicological Sciences.2019; 169(2): 409.     CrossRef
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    Silke Bergmann, Husni Elbahesh
    Virology.2019; 534: 54.     CrossRef
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    Sehee Park, Jin Il Kim, Joon-Yong Bae, Kirim Yoo, Hyunung Kim, In-Ho Kim, Man-Seong Park, Ilseob Lee
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    Zhimin Wan, Stivalis Cardenas Garcia, Jing Liu, Jefferson Santos, Silvia Carnaccini, Ginger Geiger, Lucas Ferreri, Daniela Rajao, Daniel R. Perez, Adolfo García-Sastre
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    David Ernszt, Krisztina Banfai, Zoltan Kellermayer, Attila Pap, Janet M. Lord, Judit E. Pongracz, Krisztian Kvell
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    Yufeng Song, Xiang Wang, Hongbo Zhang, Xinying Tang, Min Li, Jufang Yao, Xia Jin, Hildegund C. J. Ertl, Dongming Zhou, D. S. Lyles
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    Kannan Tharakaraman, Vidya Subramanian, Karthik Viswanathan, Susan Sloan, Hui-Ling Yen, Dale L. Barnard, Y. H. Connie Leung, Kristy J. Szretter, Tyree J. Koch, James C. Delaney, Gregory J. Babcock, Gerald N. Wogan, Ram Sasisekharan, Zachary Shriver
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    Hwan Hee Lee, Heejin Park, Gi-Ho Sung, Kanghyo Lee, Taeho Lee, Ilseob Lee, Man-seong Park, Yong Woo Jung, Yu Su Shin, Hyojeung Kang, Hyosun Cho
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    Sehee Park, Jin Il Kim, Ilseob Lee, Sangmoo Lee, Min-Woong Hwang, Joon-Yong Bae, Jun Heo, Donghwan Kim, Seok-Il Jang, Hyejin Kim, Hee Jin Cheong, Jin-Won Song, Ki-Joon Song, Luck Ju Baek, Man-Seong Park, Balaji Manicassamy
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Experimental Phasing Using Zinc and Sulfur Anomalous Signals Measured at the Zinc Absorption Peak
Sangmin Lee , Min-Kyu Kim , Chang-Jun Ji , Jin-Won Lee , Sun-Shin Cha
J. Microbiol. 2013;51(5):639-643.   Published online October 31, 2013
DOI: https://doi.org/10.1007/s12275-013-3412-2
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AbstractAbstract PDF
Iron is an essential transition metal required for bacterial growth and survival. Excess free iron can lead to the generation of reactive oxygen species that can cause severe damage to cellular functions. Cells have developed iron-sensing regulators to maintain iron homeostasis at the transcription level. The ferric uptake regulator (Fur) is an iron-responsive regulator that controls the expression of genes involved in iron homeostasis, bacterial virulence, stress resistance, and redox metabolism. Here, we report the expression, purification, crystallization, and phasing of the apo-form of Bacillus subtilis Fur (BsFur) in the absence of regulatory metal ions. Crystals were obtained by microbatch crystallization method at 295 K and diffraction data at a resolution of 2.6 Å was collected at the zinc peak wavelength (λ=1.2823 Å). Experimental phasing identified the positions of one zinc atom and four sulfur atoms of cysteine residues coordinating the zinc atom, indicating that the data contained a meaningful anomalous scattering originating from the ordered zinc-coordinating sulfur atoms, in spite of the small anomalous signals of sulfur atoms at the examined wavelength.
A Novel Retron of Vibrio parahaemolyticus Is Closely Related to Retron-Vc95 of Vibrio cholerae
Toshi Shimamoto , Ashraf M. Ahmed , Tadashi Shimamoto
J. Microbiol. 2013;51(3):323-328.   Published online June 28, 2013
DOI: https://doi.org/10.1007/s12275-013-2715-7
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AbstractAbstract PDF
Some bacteria produce a satellite RNA-DNA complex termed msDNA, multicopy single-stranded DNA. In this report, msDNA from Vibrio parahaemolyticus, a cause of acute gastroenteritis, was identified and named msDNA-Vp96. The retron element containing the ret gene, encoding the reverse transcriptase (RT) that is responsible for msDNA production, was cloned and characterized. Comparison of msDNAVp96 and msDNA-Vc95, from Vibrio cholerae, showed a high level of sequence similarity. We exchanged the two ret genes to examine whether msDNA was produced by the RT from different sources. We found that RT-Vp96 of V. parahaemolyticus was able to synthesize msDNA-Vc95 of V. cholerae and vice versa. To the best of our knowledge, this is the first report that RT from different bacterial species can synthesize msDNA.

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    Anna J Simon, Andrew D Ellington, Ilya J Finkelstein
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The Role of a Dark Septate Endophytic Fungus, Veronaeopsis simplex Y34, in Fusarium Disease Suppression in Chinese Cabbage
Rida O. Khastini , Hiroyuki Ohta , Kazuhiko Narisawa
J. Microbiol. 2012;50(4):618-624.   Published online August 25, 2012
DOI: https://doi.org/10.1007/s12275-012-2105-6
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AbstractAbstract PDF
The soil-inhabiting fungal pathogen Fusarium oxysporum has been an increasing threat to Chinese cabbage (Brassica campestris L.). A dark septate endophytic fungus, Veronaeopsis simplex Y34, isolated from Yaku Island, Japan, was evaluated in vitro for the ability to suppress Fusarium disease. Seedlings grown in the presence of the endophyte showed a 71% reduction in Fusarium wilt disease and still had good growth. The disease control was achieved through a synergetic effect involving a mechanical resistance created by a dense network of V. simplex Y34 hyphae, which colonized the host root, and siderophore production acting indirectly to induce a resistance mechanism in the plant. Changes in the relative abundance of the fungal communities in the soil as determined by fluorescently labelled T-RFs (terminal restriction fragments), appeared 3 weeks after application of the fungus. Results showed the dominance of V. simplex Y34, which became established in the rhizosphere and out-competed F. oxysporum.

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Effects of a Dark-Septate Endophytic Isolate LBF-2 on the Medicinal Plant Lycium barbarum L.
Hai-han Zhang , Ming Tang , Hui Chen , Ya-jun Wang
J. Microbiol. 2012;50(1):91-96.   Published online February 27, 2012
DOI: https://doi.org/10.1007/s12275-012-1159-9
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AbstractAbstract PDF
Dark septate endophytes (DSE) are ubiquitous root associated fungi; however, our understanding of their ecological function remains unclear. Here, we investigated the positive effect of a DSE fungus on its host plant Lycium barbarum L. A DSE isolate, LBF-2, isolated from the roots of L. barbarum, was inoculated onto the roots of plants, which were grown under greenhouse conditions for five weeks. The result of molecular analyses of internal transcribed spacer regions indicated that LBF-2 was 96% similar to Paraphoma chrysanthemicola. Melanized septate hyphae were observed in the root cortical cells of L. barbarum using a light microscope. Inoculation with LBF-2 increased the total biomass by 39.2% and also enhanced chlorophyll fluorescence. Inoculation increased the concentration of total chlorophyll by 22.8% and of chlorophyll a by 21.3%, relative to uninoculated controls. These data indicate that the LBF-2 isolate might be used to facilitate the cultivation of L. barbarum, which has medicinal applications.

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Complete Genome of Leptospirillum ferriphilum ML-04 Provides Insight into Its Physiology and Environmental Adaptation
Shuang Mi , Jian Song , Jianqun Lin , Yuanyuan Che , Huajun Zheng , Jianqiang Lin
J. Microbiol. 2011;49(6):890-901.   Published online December 28, 2011
DOI: https://doi.org/10.1007/s12275-011-1099-9
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AbstractAbstract PDF
Leptospirillum ferriphilum has been identified as the dominant, moderately thermophilic, bioleaching microorganism in bioleaching processes. It is an acidic and chemolithoautrophic bacterium that gains electrons from ferrous iron oxidation for energy production and cell growth. Genetic information about this microorganism has been limited until now, which has hindered its further exploration. In this study, the complete genome of L. ferripilum ML-04 is sequenced and annotated. The bacterium has a single circular chromosome of 2,406,157 bp containing 2,471 coding sequences (CDS), 2 rRNA operons, 48 tRNA genes, a large number of mobile genetic elements and 2 genomic islands. In silico analysis shows L. ferriphilum ML-04 fixes carbon through a reductive citric acid (rTCA) cycle, and obtains nitrogen through ammonium assimilation. The genes related to “cell envelope biogenesis, outer membrane” (6.9%) and “DNA replication, recombination and repair” (5.6%) are abundant, and a large number of genes related to heavy metal detoxification, oxidative and acidic stress defense, and signal transduction pathways were detected. The genomic plasticity, plentiful cell envelope components, inorganic element metabolic abilities and stress response mechanisms found the base for this organism’s survival in the bioleaching niche.

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Research Support, N.I.H., Extramural
Helicobacter pylori apo-Fur Regulation Appears Unconserved Across Species
Shana Miles , Beth M. Carpenter , Hanan Gancz , D. Scott Merrell
J. Microbiol. 2010;48(3):378-386.   Published online June 23, 2010
DOI: https://doi.org/10.1007/s12275-010-0022-0
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AbstractAbstract PDF
The Ferric Uptake Regulator (Fur) is a transcriptional regulator that is conserved across a broad number of bacterial species and has been shown to regulate expression of iron uptake and storage genes. Additionally, Fur has been shown to be an important colonization factor of the gastric pathogen Helicobacter pylori. In H. pylori, Fur-dependent regulation appears to be unique in that Fur is able to act as a transcriptional repressor when bound to iron as well as in its iron free (apo) form. To date, apo-regulation has not been identified in any other bacterium. To determine whether Fur from other species has the capacity for aporegulation, we investigated the ability of Fur from Escherichia coli, Campylobacter jejuni, Desulfovibrio vulgaris Hildenborough, Pseudomonas aeruginosa, and Vibrio cholerae to complement both iron-bound and apo-Fur regulation within the context of a H. pylori fur mutant. We found that while some Fur species (E. coli, C. jejuni, and V. cholerae) complemented iron-bound regulation, apo-regulation was unable to be complemented by any of the examined species. These data suggest that despite the conservation among bacterial Fur proteins, H. pylori Fur contains unique structure/function features that make it novel in comparison to Fur from other species.
Research Support, Non-U.S. Gov'ts
Experimental and Computational Characterization of the Ferric Uptake Regulator from Aliivibrio salmonicida (Vibrio salmonicida)
Hege Lynum Pedersen , Rafi Ahmad , Ellen Kristin Riise , Hanna-Kirsti Schrøder Leiros , Stefan Hauglid , Sigrun Espelid , Bjørn Olav Brandsdal , Ingar Leiros , Nils-Peder Willassen , Peik Haugen
J. Microbiol. 2010;48(2):174-183.   Published online May 1, 2010
DOI: https://doi.org/10.1007/s12275-010-9199-5
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AbstractAbstract PDF
The Ferric uptake regulator (Fur) is a global transcription factor that affects expression of bacterial genes in an iron-dependent fashion. Although the Fur protein and its iron-responsive regulon are well studied, there are still important questions that remain to be answered. For example, the consensus Fur binding site also known as the “Fur box” is under debate, and it is still unclear which Fur residues directly interact with the DNA. Our long-term goal is to dissect the biological roles of Fur in the development of the disease cold-water vibriosis, which is caused by the psychrophilic bacteria Aliivibrio salmonicida (also known as Vibrio salmonicida). Here, we have used experimental and computational methods to characterise the Fur protein from A. salmonicida (AS-Fur). Electrophoretic mobility shift assays show that AS-Fur binds to the recently proposed vibrio Fur box consensus in addition to nine promoter regions that contain Fur boxes. Binding appears to be dependent on the number of Fur boxes, and the predicted “strength” of Fur boxes. Finally, structure modeling and molecular dynamics simulations provide new insights into potential AS-Fur–DNA interactions.
Adaptation and Cross-Adaptation of Listeria monocytogenes and Salmonella enterica to Poultry Decontaminants
Alicia Alonso-Hernando , Rosa Capita , Miguel Prieto , Carlos Alonso-Calleja
J. Microbiol. 2009;47(2):142-146.   Published online May 2, 2009
DOI: https://doi.org/10.1007/s12275-008-0237-5
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AbstractAbstract PDF
Information on the potential for acquired reduced susceptibility of bacteria to poultry decontaminants occurring is lacking. Minimal Inhibitory Concentrations (MICs) were established for assessing the initial susceptibility and the adaptative and cross-adaptative responses of four bacterial strains (Listeria monocytogenes serovar 1/2a, L. monocytogenes serovar 4b, Salmonella enterica serotype Typhimurium, and S. enterica serotype Enteritidis) to four poultry decontaminants (trisodium phosphate, acidified sodium chlorite -ASC-, citric acid, and peroxyacetic acid). The initial susceptibility was observed to differ among species (all decontaminants) and between Salmonella strains (ASC). These inter- and intra-specific variations highlight (1) the need for strict monitoring of decontaminant concentrations to inactivate all target pathogens of concern, and (2) the importance of selecting adequate test strains in decontamination studies. MICs of ASC (0.17±0.02 to 0.21±0.02 mg/ml) were higher than the U.S. authorized concentration when applied as a pre-chiller or chiller solution (0.05 to 0.15 mg/ml). Progressively increasing decontaminant concentrations resulted in reduced susceptibility of strains. The highest increase in MIC was 1.88 to 2.71-fold (ASC). All decontaminants were shown to cause cross-adaptation of strains between both related and unrelated compounds, the highest increase in MIC being 1.82-fold (ASC). Our results suggest that the in-use concentrations of ASC could, in certain conditions, be ineffective against Listeria and Salmonella strains. The adaptative and cross-adaptative responses of strains tested to poultry decontaminants are of minor concern. However, the observations being presented here are based on in vitro studies, and further research into practical applications are needed in order to confirm these findings.

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Dark Septate Endophyte (DSE) Fungi Isolated from Metal Polluted Soils: Their Taxonomic Position, Tolerance, and Accumulation of Heavy Metals In Vitro
Yujie Zhang , Yan Zhang , Maojun Liu , Xiaodong Shi , Zhiwei Zhao
J. Microbiol. 2008;46(6):624-632.   Published online December 24, 2008
DOI: https://doi.org/10.1007/s12275-008-0163-6
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AbstractAbstract PDF
To understand the possible role of the plant root associated fungi on metal tolerance, their role in the uptake of heavy metals and the potential transfer of these metal ions to the plant, three strains of dark septate endophytic (DSE) fungi were isolated from a waste smelter site in southwest China, and one strain was isolated from a non-contaminated site. According to molecular phylogenetic analysis of the ITS 1-5.8S rDNA-ITS 2 gene regions and morphological characteristics, one is identified as Exophiala pisciphila, and the other three are non-sporulating fungi under the experiment condition with the nearest phylogenetic affinities to the Thysanorea papuana strain EU041814. Tolerance and accumulation abilities of the three DSE strains for metals were investigated in liquid culture. Minimum inhibitory concentrations (MIC) of Pb, Zn, and Cd were determined. It was demonstrated that the tolerance of the DSE strains varied between metal species and strains. The E. pisciphila strain is able to accumulate lead and cadmium over 20% and 5% of dry weight of biomass, respectively. Partial of the sequestrated metals can be washed with CaCl2. Morphological and enzyme activity changes taking place in the presence of excessive Pb, Cd, and/or Zn also indicate that the mechanism of heavy metal tolerance and accumulation of the DSE strains would be a complex process. The findings indicated promising tolerance and accumulation of the DSE strains with potential values in metal cycling and restoration of soil and water system.

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Cys-92, Cys-95, and the C-Terminal 12 Residues of the Vibrio harveyi Ferric Uptake Regulator (Fur) are Functionally Inessential
Kun Sun , Shuang Cheng , Min Zhang , Fang Wang , Li Sun
J. Microbiol. 2008;46(6):670-680.   Published online December 24, 2008
DOI: https://doi.org/10.1007/s12275-008-0113-3
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AbstractAbstract PDF
Ferric uptake regulator (Fur) is a global regulator involved in multiple aspects of bacterial life. The gene encoding the Vibrio harveyi Fur (FurVh) was cloned from a pathogenic V. harveyi strain isolated from diseased fish. FurVh shares 77% overall sequence identity with the Escherichia coli Fur (FurEc) and could complement a mutant of FurEc. Like FurEc, FurVh possesses two cysteine residues at positions 92 and 95, yet unlike FurEc, in which these cysteine residues constitute part of the metal ion coordination site and hence are vital to the repressor activity, C92 and C95 of FurVh proved to be functionally inessential. Further study identified a Vibrio Fur signature sequence, which is preserved in all the ten Vibrio Fur proteins that have been discovered to date but in none of the non-vibrio Fur proteins. Site-directed and random mutation analyses of the signature residues, the cysteine residues, and seven highly charged amino acid residues indicated that D9, H32, C137, and K138 of FurVh are functionally important but D9, C137, and K138 can be replaced by more than one functional substitutes. Systematic deletion analysis demonstrated that the C-terminal 12 residues of FurVh are functionally inessential. These results (i) indicated that the activation mechanism, or certain aspects of which, of FurVh is possibly different from that of FurEc; and (ii) suggested that it is not very likely that the C-terminal 12 residues play any significant role in the activation or stability of FurVh; and (iii) provided insights into the potential function of the local structure involving C137 and K138.

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    Shana Miles, Beth M. Carpenter, Hanan Gancz, D. Scott Merrell
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Phylogenetic Diversity of Acidophilic Sporoactinobacteria Isolated from Various Soils
Sung-Heun Cho , Ji-Hye Han , Chi Nam Seong , Seung Bum Kim
J. Microbiol. 2006;44(6):600-606.
DOI: https://doi.org/2468 [pii]
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Spore forming actinobacteria (sporoactinobacteria) isolated from soils with an acidic pH in Pinus thunbergii forests and coal mine waste were subjected to taxonomic characterization. For the isolation of acidophilic actinobacteria, acidified starch casein agar (pH adjusted to 4-5) was used. The numbers of actinobacteria growing in acidic media were between 3.2 × 104 and 8.0 × 106 CFU/g soil. Forty three acidophilic actinobacterial strains were isolated and their 16S rDNA sequences were determined. The isolates were divided into eight distinctive phylogenetic clusters within the variation encompassed by the family Streptomycetaceae. Four clusters among them were assigned to the genus Streptacidiphilus, whereas the remaining four were assigned to Streptomyces. The clusters belonging to either Streptomyces or Streptacidiphilus did not form monophyletic clade. The growth pH profiles indicated that the representative isolates grew best between pH 5 and 6. It is evident from this study that acidity has played a critical role in the differentiation of the family Streptomycetaceae, and also that different mechanisms might have resulted in the evolution of two groups, Streptacidiphilus (strict acidophiles) and neutrotolerant acidophilic Streptomyces. The effect of geographic separation was clearly seen among the Streptacidiphilus isolates, which may be a key factor in speciation of the genus.
Identification and Expression of the cym, cmt, and tod Catabolic Genes from Pseudomonas putida KL47: Expression of the Regulatory todST Genes as a Factor for Catabolic Adaptation
Kyoung Lee , Eun Kyeong Ryu , Kyung Soon Choi , Min Chul Cho , Jae Jun Jeong , Eun Na Choi , Soo O Lee , Do-Young Yoon , Ingyu Hwang , Chi-Kyung Kim
J. Microbiol. 2006;44(2):192-199.
DOI: https://doi.org/2365 [pii]
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AbstractAbstract PDF
Pseudomonas putida KL47 is a natural isolate that assimilates benzene, 1-alkylbenzene (C1-C4), biphenyl, p-cumate, and p-cymene. The genetic background of strain KL47 underlying the broad range of growth substrates was examined. It was found that the cym and cmt operons are constitutively expressed due to a lack of the cymR gene, and the tod operon is still inducible by toluene and biphenyl. The entire array of gene clusters responsible for the catabolism of toluene and p-cymene/p-cumate has been cloned in a cosmid vector, pLAFR3, and were named pEK6 and pEK27, respectively. The two inserts overlap one another and the nucleotide sequence (42,505 bp) comprising the cym, cmt, and tod operons and its flanking genes in KL47 are almost identical (>99%) to those of P. putida F1. In the cloned DNA fragment, two genes with unknown functions, labeled cymZ and cmtR, were newly identified and show high sequence homology to dienelactone hydrolase and CymR proteins, respectively. The cmtR gene was identified in the place of the cmtI gene of previous annotation. Western blot analysis showed that, in strains F1 and KL47, the todT gene is not expressed during growth on Luria Bertani medium. In minimal basal salt medium, expression of the todT gene is inducible by toluene, but not by biphenyl in strain F1; however, it is constantly expressed in strain KL47, indicating that high levels of expression of the todST genes with one amino acid substitution in TodS might provide strain KL47 with a means of adaptation of the tod catabolic operon to various aromatic hydrocarbons.
A Proteomic Approach to Study msDNA Function in Escherichia coli
Mi-Ae Jeong , Dongbin Lim
J. Microbiol. 2004;42(3):200-204.
DOI: https://doi.org/2089 [pii]
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AbstractAbstract PDF
Retron is a prokaryotic genetic element that produces multicopy single-stranded DNA covalently linked to RNA (msDNA) by a reverse transcriptase. It was found that cells producing a large amount of msDNA, rather than those that did not, showed a higher rate of mutation. In order to understand the molecular mechanism connecting msDNA production to the high mutation rate the protein patterns were compared by two dimensional gel electrophoresis. Ten proteins were found to be differentially expressed at levels more than three fold greater in cells with than without msDNA, nine of which were identified by MALDI TOF MS. Eight of the nine identified proteins were repressed in msDNA-producing cells and, surprisingly, most were proteins functioning in the dissimilation of various carbon sources. One protein was induced four fold greater in the msDNA producing cells and was identified as a 30S ribosomal protein S2 involved in the regulation of translation. The molecular mechanism underlying the elevated mutation in msDNA-producing cell still remains elusive.
Review
The Use of the Rare UUA Codon to Define "Expression Space" for Genes Involved in Secondary Metabolism, Development and Environmental Adaptation in Streptomyces
Keith F. Chater , Govind Chandra
J. Microbiol. 2008;46(1):1-11.
DOI: https://doi.org/10.1007/s12275-007-0233-1
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AbstractAbstract PDF
In Streptomyces coelicolor, bldA encodes the only tRNA for a rare leucine codon, UUA. This tRNA is unnecessary for growth, but is required for some aspects of secondary metabolism and morphological development, as revealed by the phenotypes of bldA mutants in diverse streptomycetes. This article is a comprehensive review of out understanding of this unusual situation. Based on information from four sequenced genomes it now appears that, typically, about 2~3% of genes in any one streptomycete contain a TTA codon, most having been acquired through species-specific horizontal gene transfer. Among the few widely conserved TTA-containing genes, mutations in just one, the pleiotropic regulatory gene adpA, give an obvious phenotype: such mutants are defective in aerial growth and sporulation, but vary in the extent of their impairment in secondary metabolism in different streptomycetes. The TTA codon in adpA is largely responsible for the morphological phenotype of a bldA mutant of S. coelicolor. AdpA-dependent targets include several genes involved in the integrated action of extracellular proteases that, at least in some species, are involved in the conversion of primary biomass into spores. The effects of bldA mutations on secondary metabolism are mostly attributable to the presence of TTA codons in pathway-specific genes, particularly in transcriptional activator genes. This is not confined to S. coelicolor-it is true for about half of all known antibiotic biosynthetic gene sets from streptomycetes. Combined microarray and proteomic analysis of liquid (and therefore non-sporulating) S. coelicolor bldA mutant cultures revealed effects of the mutation during rapid growth, during transition phase, and in stationary phase. Some of these effects may be secondary consequences of changes in the pattern of ppGpp accumulation. It is argued that the preferential accumulation of the bldA tRNA under conditions in which growth is significantly constrained has evolved to favour the expression of genes that confer adaptive benefits in intermittently encountered sub-optimal environments. The evolution of this system may have been a secondary consequence of the selective pressure exerted by bacteriophage attack. Some biotechnological implications of bldA phenomenology are considered.

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Incorpotation and production of glucose in Lake Soyang
kwag, No Tae , Choi, Seung Ik , Ahn, Tae Young , Ahn, Tae Seok
J. Microbiol. 1995;33(1):74-79.
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AbstractAbstract PDF
Kinetics of heterotrophic activity (glucose uptake) and extracellular enzyme activity(β-glucosidase, cellobiohydrolase) and cell numbers were measured in Lake Soyang during phytoplankton bloom development and after its breakdown. V_max for glucose was lower during Diatom bloom and that was higher after its breakdown. But the increase ion β-glucosidase activity was detected in late of Diatom bloom. Glucose uptake did not associated with β-glucosidase activity. The tight relationship between β-glucosidase and the incorporation of glucose by bacteria was not shown and the significance of depolymerization on the incorporation of glucose in lake water are discussed.

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