Journal of Microbiology

Journal Articles

Transcription Factors Tec1 and Tec2 Play Key Roles in the Hyphal Growth and Virulence of Mucor lusitanicus Through Increased Mitochondrial Oxidative Metabolism: Viridiana Alejandre-Castañeda , J. Alberto Patiño-Medina , Marco I. Valle-Maldonado , Alexis García , Rafael Ortiz-Alvarado , León F. Ruíz-Herrera , Karla Viridiana Castro-Cerritos , Joel Ramírez-Emiliano , Martha I. Ramírez-Díaz , Victoriano Garre , Soo Chan Lee , Víctor Meza-Carmen; J. Microbiol. 2023;61(12):1043-1062. Published online December 19, 2023; DOI: https://doi.org/10.1007/s12275-023-00096-8

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Mucormycosis is a lethal and difficult-to-treat fungal infection caused by fungi of the order Mucorales. Mucor lusitanicus, a member of Mucorales, is commonly used as a model to understand disease pathogenesis. However, transcriptional control of hyphal growth and virulence in Mucorales is poorly understood. This study aimed to investigate the role of Tec proteins, which belong to the TEA/ATTS transcription factor family, in the hyphal development and virulence of M. lusitanicus. Unlike in the genome of Ascomycetes and Basidiomycetes, which have a single Tec homologue, in the genome of Mucorales, two Tec homologues, Tec1 and Tec2, were found, except in that of Phycomyces blakesleeanus, with only one Tec homologue. tec1 and tec2 overexpression in M. lusitanicus increased mycelial growth, mitochondrial content and activity, expression of the rhizoferrin synthetase-encoding gene rfs, and virulence in nematodes and wax moth larvae but decreased cAMP levels and protein kinase A (PKA) activity. Furthermore, tec1- and tec2-overexpressing strains required adequate mitochondrial metabolism to promote the virulent phenotype. The heterotrimeric G beta subunit 1-encoding gene deletant strain (Δgpb1) increased cAMP-PKA activity, downregulation of both tec genes, decreased both virulence and hyphal development, but tec1 and tec2 overexpression restored these defects. Overexpression of allele-mutated variants of Tec1(S332A) and Tec2(S168A) in the putative phosphorylation sites for PKA increased both virulence and hyphal growth of Δgpb1. These findings suggest that Tec homologues promote mycelial development and virulence by enhancing mitochondrial metabolism and rhizoferrin accumulation, providing new information for the rational control of the virulent phenotype of M. lusitanicus.

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A one-pot five component reaction for the synthesis of tetrazol-benzofuran hybrids and their inhibitory activity against Mucor lusitanicus
Cesia M. Aguilar-Morales, Viridiana Alejandre-Castañeda, Claudia Contreras-Celedón, Martha Isela Ramírez-Díaz, Alejandro Islas-Jácome, Victor Meza-Carmen, Luis Chacón-García, Carlos J. Cortés-García
Organic & Biomolecular Chemistry.2024; 22(35): 7240. CrossRef
Functional characterization of two survival factor 1 genes in Mucor lusitanicus
Olivér Jáger, Csilla Szebenyi, Tammam Khaliefeh Siliman Abu Saleem, Anna Molnár, Vanda Kovács, Karina Kiss, Mónika Homa, Bernadett Vágó, Sándor Kiss-Vetráb, Mónika Varga, Rita Sinka, Csaba Vágvölgyi, Gábor Nagy, Tamás Papp, Renato Kovacs
Microbiology Spectrum.2024;[Epub] CrossRef

Crystal structure of the phage-encoded N-acetyltransferase in complex with acetyl-CoA, revealing a novel dimeric arrangement: Nayeon Ki , Inseong Jo , Yongseong Hyun , Jinwook Lee , Nam-Chul Ha , Hyun-Myung Oh; J. Microbiol. 2022;60(7):746-755. Published online July 4, 2022; DOI: https://doi.org/10.1007/s12275-022-2030-2

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Abstract

Bacteriophages employ diverse mechanisms to facilitate the proliferation of bacteriophages. The Salmonella-infecting phage SPN3US contains a putative N-acetyltransferase, which is widely found in bacteriophages. However, due to low sequence similarity to the N-acetyltransferases from bacteria and eukaryotic cells, the structure and function of phage-encoded acetyltransferases are mainly unknown. This study determines the crystal structure of the putative N-acetyltransferase of SPN3US in complex with acetyl-CoA. The crystal structure showed a novel homodimeric arrangement stabilized by exchanging the C-terminal α-helix within the dimer. The following biochemical analyses suggested that the phageencoded acetyltransferase might have a very narrow substrate specificity. Further studies are required to reveal the biochemical activity, which would help elucidate the interaction between the phage and host bacteria in controlling pathogenic bacteria.

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Posttranslational modifications in bacteria during phage infection
Hannelore Longin, Nand Broeckaert, Vera van Noort, Rob Lavigne, Hanne Hendrix
Current Opinion in Microbiology.2024; 77: 102425. CrossRef

Antiviral effects of human placenta hydrolysate (Laennec) against SARS-CoV-2 in vitro and in the ferret model: Eun-Ha Kim , Young-il Kim , Seung-Gyu Jang , Minju Im , Kyeongsoo Jeong , Young Ki Choi , Hae-Jung Han; J. Microbiol. 2021;59(11):1056-1062. Published online October 6, 2021; DOI: https://doi.org/10.1007/s12275-021-1367-2

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Abstract

The COVID-19 pandemic has caused unprecedented health, social, and economic crises worldwide. However, to date, there is an only a limited effective treatment for this disease. Human placenta hydrolysate (hPH) has previously been shown to be safe and to improve the health condition in patients with hyperferritinemia and COVID-19. In this study, we aimed to determine the antiviral effects of hPH against SARS-CoV-2 in vitro and in vivo models and compared with Remdesivir, an FDA-approved drug for COVID-19 treatment. To assess whether hPH inhibited SARS-CoV-2 replication, we determined the CC50, EC50, and selective index (SI) in Vero cells by infection with a SARS-CoV-2 at an MOI of 0.01. Further, groups of ferrets infected with 105.8 TCID50/ml of SARS-CoV-2 and treated with hPH at 2, 4, 6 dpi, and compared their clinical manifestation and virus titers in respiratory tracts with PBS control-treated group. The mRNA expression of immunerelated cytokines was determined by qRT-PCR. hPH treatment attenuated virus replication in a dose-dependent manner in vitro. In a ferret infection study, treatment with hPH resulted in minimal bodyweight loss and attenuated virus replication in the nasal wash, turbinates, and lungs of infected ferrets. In addition, qRT-PCR results revealed that the hPH treatment remarkably upregulated the gene expression of type I (IFN-α and IFN-β) and II (IFN-γ) IFNs in SARS-CoV-2 infected ferrets. Our data collectively suggest that hPH has antiviral efficacy against SARS-CoV-2 and might be a promising therapeutic agent for the treatment of SARS-CoV-2 infection.

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Perinatal Hypoxia and Immune System Activation in Schizophrenia Pathogenesis: Critical Considerations During COVID-19 Pandemic
I Kawikova, K Hakenova, M Lebedeva, L Kleteckova, L Jakob, V Spicka, L Wen, F Spaniel, K Vales
Physiological Research.2024; : S615. CrossRef
Human Placenta Extract (HPH) Suppresses Inflammatory Responses in TNF-α/IFN-γ-Stimulated HaCaT Cells and a DNCB Atopic Dermatitis (AD)-Like Mouse Model
Jung Ok Lee, Youna Jang, A Yeon Park, Jung Min Lee, Kyeongsoo Jeong, So-Hyun Jeon, Hui Jin, Minju Im, Jae-Won Kim, Beom Joon Kim
Journal of Microbiology and Biotechnology.2024; 34(10): 1969. CrossRef
Systematic analysis of the pharmacology of standardized extracts of human placenta
T. E. Bogacheva, I. Yu. Torshin, O. A. Gromova
Pharmacokinetics and Pharmacodynamics.2024; (4): 3. CrossRef
Distinctive Combinations of RBD Mutations Contribute to Antibody Evasion in the Case of the SARS-CoV-2 Beta Variant
Tae-Hun Kim, Sojung Bae, Sunggeun Goo, Jinjong Myoung
Journal of Microbiology and Biotechnology.2023; 33(12): 1587. CrossRef
Current state-of-the-art and potential future therapeutic drugs against COVID-19
Ailong Sha, Yi Liu, Haiyan Hao
Frontiers in Cell and Developmental Biology.2023;[Epub] CrossRef
SARS-CoV-2 Aerosol and Intranasal Exposure Models in Ferrets
Elizabeth E. Zumbrun, Samantha E. Zak, Eric D. Lee, Philip A. Bowling, Sara I. Ruiz, Xiankun Zeng, Jeffrey W. Koehler, Korey L. Delp, Russel R. Bakken, Shannon S. Hentschel, Holly A. Bloomfield, Keersten M. Ricks, Tamara L. Clements, April M. Babka, John
Viruses.2023; 15(12): 2341. CrossRef
Human placenta hydrolysates: from V.P. Filatov to the present day: Review
Olga A. Gromova, Ivan Yu. Torshin, Alexander G. Chuchalin, Valeriy А. Maximov
Terapevticheskii arkhiv.2022; 94(3): 434. CrossRef

Description of Ornithinimicrobium ciconiae sp. nov., and Ornithinimicrobium avium sp. nov., isolated from the faeces of the endangered and near-threatened birds: So-Yeon Lee , Hojun Sung , Pil Soo Kim , Hyun Sik Kim , Jae-Yun Lee , June-Young Lee , Yun-Seok Jeong , Euon Jung Tak , Jeong Eun Han , Dong-Wook Hyun , Jin-Woo Bae; J. Microbiol. 2021;59(11):978-987. Published online September 27, 2021; DOI: https://doi.org/10.1007/s12275-021-1323-1

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Abstract

Phenotypic and genomic analyses were performed to characterize two novel species, H23M54T and AMA3305T, isolated from the faeces of the Oriental stork (Ciconia boyciana) and the cinereous vulture (Aegypius monachus), respectively. Strains H23M54T and AMA3305T showed the highest similarities of 16S rRNA gene sequences and complete genome sequences with Ornithinimicrobium cavernae CFH 30183T (98.5% of 16S rRNA gene sequence similarity and 82.1% of average nucleotide identity, ANI) and O. pekingense DSM 21552T (98.5% of 16S rRNA gene sequence similarity and 82.3% of ANI), respectively. Both strains were Gram-stain-positive, obligate aerobes, non-motile, non-spore-forming, and coccoid- and rodshaped. Strain H23M54T grew optimally at 25–30°C and pH 8.0 and in the presence of 1.5–2% (wt/vol) NaCl, while strain AMA3305T grew optimally at 30°C and pH 7.0 and in the presence of 1–3% (wt/vol) NaCl. Both strains had iso-C15:0, iso- C16:0, and summed feature 9 (iso-C17:1 ω9c and/or C16:0 10- methyl) as major cellular fatty acids. MK-8 (H4) was identified as the primary respiratory quinone in both strains. Strains H23M54T and AMA3305T possessed diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. Moreover, strains H23M54T and AMA3305T commonly contained ribose and glucose as major sugars and L-ornithine, L-alanine, glycine, and aspartic acid as major amino acids. The polyphasic taxonomic data indicate that strains H23M54T and AMA3305T represent novel species of the genus Ornithinimicrobium. We propose the names Ornithinimicrobium ciconiae sp. nov. and Ornithinimicrobium avium sp. nov. for strains H23M54T (= KCTC 49151T = JCM 33221T) and AMA3305T (= KCTC 49180T = JCM 32873T), respectively.

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Molecular insight and antimicrobial potential of Actinomycetota isolated from Tanzania’s seagrass sediments
Lucy Dalusi Mbusi, Sylvester Leonard Lyantagaye, Thomas Jacob Lyimo
Biologia.2024; 80(1): 163. CrossRef
Bacterial community of ticks (Acari: Ixodidae) and mammals from Arauca, Colombian Orinoquia
Paula A. Ossa-López, Héctor E. Ramírez-Chaves, María Elena Álvarez López, Gabriel Jaime Castaño Villa, Fredy A. Rivera-Páez
International Journal for Parasitology: Parasites and Wildlife.2024; 24: 100943. CrossRef
Morphological and genomic characteristics of two novel actinomycetes, Ornithinimicrobium sufpigmenti sp. nov. and Ornithinimicrobium faecis sp. nov. isolated from bat faeces (Rousettus leschenaultia and Taphozous perforates)
Yuyuan Huang, Suping Zhang, Yuanmeihui Tao, Jing Yang, Shan Lu, Dong Jin, Ji Pu, Wenbo Luo, Han Zheng, Liyun Liu, Jia-fu Jiang, Jianguo Xu
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Journal of Microbiology.2023; 61(4): 379. CrossRef
An update on novel taxa and revised taxonomic status of bacteria isolated from non-domestic animals described in 2022
Claire R. Burbick, Sara D. Lawhon, Erik Munson, Elizabeth Thelen, Amanda Zapp, Anastasia Wilson, Romney M. Humphries
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Valid publication of new names and new combinations effectively published outside the IJSEM
Aharon Oren, George M. Garrity
International Journal of Systematic and Evolutionary Microbiology .2022;[Epub] CrossRef
Lysobacter ciconiae sp. nov., and Lysobacter avium sp. nov., isolated from the faeces of an Oriental stork
So-Yeon Lee, Pil Soo Kim, Hojun Sung, Dong-Wook Hyun, Jin-Woo Bae
Journal of Microbiology.2022; 60(5): 469. CrossRef

FgIlv3a is crucial in branched-chain amino acid biosynthesis, vegetative differentiation, and virulence in Fusarium graminearum: Xin Liu , Yichen Jiang , Yinghui Zhang , Mingzheng Yu , Hongjun Jiang , Jianhong Xu , Jianrong Shi; J. Microbiol. 2019;57(8):694-703. Published online May 11, 2019; DOI: https://doi.org/10.1007/s12275-019-9123-6

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Abstract

Dihydroxyacid dehydratase (DHAD), encoded by ILV3, catalyses the third step in the biosynthetic pathway of branchedchain amino acids (BCAAs), which include isoleucine (Ile), leucine (Leu), and valine (Val). Enzymes involved in BCAA biosynthesis exist in bacteria, plants, and fungi but not in mammals and are therefore attractive targets for antimicrobial or herbicide development. In this study, three paralogous ILV3 genes (FgILV3A, FgILV3B, and FgILV3C) were identified in the genome of Fusarium graminearum, the causal agent of Fusarium head blight (FHB). Deletion of FgILV3A alone or combined with FgILV3B or FgILV3C indicated an important role for FgILV3A in BCAA biosynthesis. FgILV3A deletion mutants lost the ability to grow on medium lacking amino acids. Exogenous supplementation of 1 mM Ile and Val rescued the auxotrophy of ΔFgIlv3A, though 5 mM was required to recover the growth defects in ΔFgIlv3AB and ΔFgIlv3AC strains, indicating that FgIlv3b and FgIlv3c exhibit redundant but accessory roles with FgIlv3a in BCAA biosynthesis. The auxotrophy of ΔFgIlv3A resulted in pleiotropic defects in aerial hyphal growth, in conidial formation and germination, and in aurofusarin accumulation. In addition, the mutants showed reduced virulence and deoxynivalenol production. Overall, our study demonstrates that FgIlv3a is crucial for BCAA biosynthesis in F. graminearum and a candidate fungicide target for FHB management.

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AflaILVB/G/I and AflaILVD are involved in mycelial production, aflatoxin biosynthesis, and fungal virulence in Aspergillus flavus
Yarong Zhao, Chulan Huang, Rui Zeng, Peirong Chen, Kaihang Xu, Xiaomei Huang, Xu Wang
Frontiers in Cellular and Infection Microbiology.2024;[Epub] CrossRef
Histone H3 N-Terminal Lysine Acetylation Governs Fungal Growth, Conidiation, and Pathogenicity through Regulating Gene Expression in Fusarium pseudograminearum
Hang Jiang, Lifang Yuan, Liguo Ma, Kai Qi, Yueli Zhang, Bo Zhang, Guoping Ma, Junshan Qi
Journal of Fungi.2024; 10(6): 379. CrossRef
Identification and Characterization of an Antifungal Gene Mt1 from Bacillus subtilis by Affecting Amino Acid Metabolism in Fusarium graminearum
Pei Song, Wubei Dong
International Journal of Molecular Sciences.2023; 24(10): 8857. CrossRef
Branched-chain amino acid biosynthesis in fungi
Gary Jones, Jane Usher, Joel T. Steyer, Richard B. Todd
Essays in Biochemistry.2023; 67(5): 865. CrossRef
FgLEU1 Is Involved in Leucine Biosynthesis, Sexual Reproduction, and Full Virulence in Fusarium graminearum
Shaohua Sun, Mingyu Wang, Chunjie Liu, Yilin Tao, Tian Wang, Yuancun Liang, Li Zhang, Jinfeng Yu
Journal of Fungi.2022; 8(10): 1090. CrossRef
Acetolactate synthases regulatory subunit and catalytic subunit genes VdILVs are involved in BCAA biosynthesis, microscletotial and conidial formation and virulence in Verticillium dahliae
ShengNan Shao, Biao Li, Qi Sun, PeiRu Guo, YeJuan Du, JiaFeng Huang
Fungal Genetics and Biology.2022; 159: 103667. CrossRef
Molecular targets for antifungals in amino acid and protein biosynthetic pathways
Aleksandra Kuplińska, Kamila Rząd
Amino Acids.2021; 53(7): 961. CrossRef
MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection of Magnaporthe oryzae
Osakina Aron, Min Wang, Anjago Wilfred Mabeche, Batool Wajjiha, Meiqin Li, Shuai Yang, Haixia You, Yan Cai, Tian Zhang, Yunxi Li, Baohua Wang, Dongmei Zhang, Zonghua Wang, Wei Tang
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Marta J. Fiołka, Paulina Czaplewska, Sylwia Wójcik-Mieszawska, Aleksandra Lewandowska, Kinga Lewtak, Weronika Sofińska-Chmiel, Tomasz Buchwald
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The pyruvate dehydrogenase kinase 2 (PDK2) is associated with conidiation, mycelial growth, and pathogenicity in Fusarium graminearum
Tao Gao, Dan He, Xin Liu, Fang Ji, Jianhong Xu, Jianrong Shi
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The Intermediates in Branched-Chain Amino Acid Biosynthesis Are Indispensable for Conidial Germination of the Insect-Pathogenic Fungus Metarhizium robertsii
Feifei Luo, Hongxia Zhou, Xue Zhou, Xiangyun Xie, You Li, Fenglin Hu, Bo Huang, Karyn N. Johnson
Applied and Environmental Microbiology.2020;[Epub] CrossRef

Identification and heterologous reconstitution of a 5-alk(en)ylresorcinol synthase from endophytic fungus Shiraia sp. Slf14: Huiwen Yan , Lei Sun , Jinge Huang , Yixing Qiu , Fuchao Xu , Riming Yan , Du Zhu , Wei Wang , Jixun Zhan; J. Microbiol. 2018;56(11):805-812. Published online October 24, 2018; DOI: https://doi.org/10.1007/s12275-018-8278-x

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Abstract

A new type III polyketide synthase gene (Ssars) was discovered from the genome of Shiraia sp. Slf14, an endophytic fungal strain from Huperzia serrata. The intron-free gene was cloned from the cDNA and ligated to two expression vectors pET28a and YEpADH2p-URA3 for expression in Escherichia coli BL21(DE3) and Saccharomyces cerevisiae BJ5464, respectively. SsARS was efficiently expressed in E. coli BL21(DE3), leading to the synthesis of a series of polyketide products. Six major products were isolated from the engineered E. coli and characterized as 1,3-dihydroxyphenyl- 5-undecane, 1,3-dihydroxyphenyl-5-cis-6􍿁-tridecene,1,3-dihydroxyphenyl- 5-tridecane, 1,3-dihydroxyphenyl-5-cis-8􍿁- pentadecene, 1,3-dihydroxyphenyl-5-pentadecane, and 1,3- dihydroxyphenyl-5-cis-10􍿁-heptadecene, respectively, based on the spectral data and biosynthetic origin. Expression of SsARS in the yeast also led to the synthesis of the same polyketide products, indicating that this enzyme can be reconstituted in both heterologous hosts. Supplementation of soybean oil into the culture of E. coli BL21(DE3)/SsARS increased the production titers of 1–6 and led to the synthesis of an additional product, which was identified as 5-(8􍿁Z,11􍿁Z-heptadecadienyl) resorcinol. This work thus allowed the identification of SsARS as a 5-alk(en)ylresorcinol synthase with flexible substrate specificity toward endogenous and exogenous fatty acids. Desired resorcinol derivatives may be synthesized by supplying corresponding fatty acids into the culture medium.

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Lorena Resende Oliveira, Ariany Rosa Gonçalves, Eliane Dias Quintela, Leandro Colognese, Marcio Vinicius de C. Barros Cortes, Marta Cristina Corsi de Filippi
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Evaluation of the Antimicrobial Potency of Silver Nanoparticles Biosynthesized by Using an Endophytic Fungus, Cryptosporiopsis ericae PS4: Lamabam Sophiya Devi , Santa Ram Joshi; J. Microbiol. 2014;52(8):667-674. Published online July 4, 2014; DOI: https://doi.org/10.1007/s12275-014-4113-1

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Abstract

In the present study, silver nanoparticles (AgNPs) with an average particle size of 5.5 ± 3.1 nm were biosynthesized using an endophytic fungus Cryptosporiopsis ericae PS4 isolated from the ethno-medicinal plant Potentilla fulgens L. The nanoparticles were characterized using UV-visible spectrophotometer, transmission electron microscopy (TEM), scanning electron microscopy (SEM), selective area electron diffraction (SAED), and energy dispersive X-ray (EDX) spectroscopy analysis. Antimicrobial efficacy of the AgNPs was analyzed singly and in combination with the antibiotic/antifungal agent chloramphenicol/fluconazole, against five pathogenic microorganisms-Staphylococcus aureus MTCC96, Salmonella enteric MTCC735, Escherichia coli MTCC730, Enterococcus faecalis MTCC2729, and Candida albicans MTCC 183. The activity of AgNPs on the growth and morphology of the microorganisms was studied in solid and liquid growth media employing various susceptibility assays. These studies demonstrated that concentrations of AgNPs alone between 10 and 25 μM reduced the growth rates of the tested bacteria and fungus and revealed bactericidal/fungicidal activity of the AgNPs by delaying the exponential and stationary phases. Examination using SEM showed pits and ruptures in bacterial cells indicating fragmented cell membrane and severe cell damage in those cultures treated with AgNPs. These experimental findings suggest that the biosynthesized AgNPs may be a potential antimicrobial agent.

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Journal Article

Application of Statistical Experimental Design for Optimization of Silver Nanoparticles Biosynthesis by a Nanofactory Streptomyces viridochromogenes: Noura El-Ahmady El-Naggar , Nayera A.M. Abdelwahed; J. Microbiol. 2014;52(1):53-63. Published online January 4, 2014; DOI: https://doi.org/10.1007/s12275-014-3410-z

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Abstract

Central composite design was chosen to determine the combined effects of four process variables (AgNO3 concentration, incubation period, pH level and inoculum size) on the extracellular biosynthesis of silver nanoparticles (AgNPs) by Streptomycesviridochromogenes. Statistical analysis of the results showed that incubation period, initial pH level and inoculum size had significant effects (P􌥑0.05) on the biosynthesis of silver nanoparticles at their individual level. The maximum biosynthesis of silver nanoparticles was achieved at a concentration of 0.5% (v/v) of 1 mM AgNO3, incubation period of 96 h, initial pH of 9 and inoculum size of 2% (v/v). After optimization, the biosynthesis of silver nanoparticles was improved by approximately 5-fold as compared to that of the unoptimized conditions. The synthetic process of silver nanoparticle generation using the reduction of aqueous Ag+ ion by the culture supernatants of S. viridochromogenes was quite fast, and silver nanoparticles were formed immediately by the addition of AgNO3 solution (1 mM) to the cell-free supernatant. Initial characterization of silver nanoparticles was performed by visual observation of color change from yellow to intense brown color. UV-visible spectrophotometry for measuring surface plasmon resonance showed a single absorption peak at 400 nm, which confirmed the presence of silver nanoparticles. Fourier Transform Infrared Spectroscopy analysis provided evidence for proteins as possible reducing and capping agents for stabilizing the nanoparticles. Transmission Electron Microscopy revealed the extracellular formation of spherical silver nanoparticles in the size range of 2.15–7.27 nm. Compared to the cell-free supernatant, the biosynthesized AgNPs revealed superior antimicrobial activity against Gram-negative, Gram-positive bacterial strains and Candida albicans.

Citations

Citations to this article as recorded by

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Research Support, Non-U.S. Gov't

Functional Characterization of the Genes tauO, tauK, and tauI in the Biosynthesis of Tautomycetin: Fen Wang , Rixiang Kong , Bo Liu , Jing Zhao , Rongguo Qiu , Li Tang; J. Microbiol. 2012;50(5):770-776. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2154-x

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Abstract: Tautomycetin is a specific protein phosphatase I inhibitor. In an effort to elucidate the biosynthetic mechanism of tautomycetin, we inactivated genes of the tautomycetin biosynthetic gene cluster, tauI, tauO, and tauK, which encode for putative P450 oxidase, citryl-CoA lyase, and esterase enzymes, respectively. The mutant STQ0606 (ΔtauO) did not produce any detectable amount of tautomycetin intermediates but could convert dialkylmaleic anhydride to tautomycetin, strongly indicating that TauO was involved in dialkylmaleic anhydride biosynthesis. STQ1211 (ΔtauK) accumulated dialkylmaleic anhydride, whereas the cofermentation of STQ1211 (ΔtauK) and STQ0606 (ΔtauO) restored the production of tautomycetin. Together, these results suggest that TauK was responsible for the conjugation of dialkylmaleic anhydride and the polyketide moiety in tautomycetin biosynthesis. The disruption of tauI resulted in the accumulation of 5-des-ketotautomycetin, revealing that TauI was responsible for the oxidation at C5 as the last step. Although the shunt pathways were involved in the biosynthesis of tautomycetin, the main post-polyketide synthase tailoring steps were dehydration, decarboxylation and oxidation, taking place consecutively. This study allowed us to predict the biosynthesis of tautomycetin more accurately and provided novel insights into the mechanism of the biosynthesis of tautomycetin.

Research Support, U.S. Gov't, Non-P.H.S.

Transcriptional Control of Genes Involved in Yeast Phospholipid Biosynthesis: Roshini Wimalarathna , Chen-Han Tsai , Chang-Hui Shen; J. Microbiol. 2011;49(2):265-273. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-1130-1

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Abstract: Phospholipid biosynthetic genes encode enzymes responsible for phospholipid biosynthesis. They are coordinately regulated by the availability of phospholipid precursors through the inositol-sensitive upstream activating sequence (UASINO). However, not all phospholipid genes are UASINO-containing genes and not all UASINO-containing genes have the same response to the phospholipid precursors. Therefore, the transcriptional regulation of phospholipid genes in response to the availability of phospholipid precursors is still unclear. Here, 22 out of 47 phospholipid biosynthetic genes were identified as UASINO-containing genes, including EKI1, EPT1, INM1, IPK2, KCS1, PAH1, and PIK1 which have never been reported before. We also showed, using qRTPCR technique, that 12 UASINO-containing genes are down-regulated by 100 μM inositol in the wild type cells and up-regulated by 100 μM inositol in the ino2Δ cells. Therefore, it is possible that these genes are transcriptionally regulated by the UASINO through the negative response of Ino2p to inositol. One other UASINO-containing gene might be regulated by the positive response of Ino2p to 100 μM inositol. Surprisingly, we found 9 UASINO-containing genes are not dependent on the response of Ino2p to 100 μM inositol, indicating that they may be regulated by other pathway. Furthermore, we identified 9 and 3 non-UASINO-containing genes that are possibly regulated by the negative and positive response of Ino2p to 100 μM inositol, respectively. Therefore, these observations provide insight into the understanding of the co-regulated phospholipid biosynthetic genes expression.

Research Support, Non-U.S. Gov'ts

Altered Protein Expression Patterns of Mycobacterium tuberculosis Induced by ATB107: Hongbo Shen , Enzhuo Yang , Feifei Wang , Ruiliang Jin , Shengfeng Xu , Qiang Huang , Honghai Wang; J. Microbiol. 2010;48(3):337-346. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-010-9315-6

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Abstract: ATB107 is a potent inhibitor of indole-3-glycerol phosphate synthase (IGPS). It can effectively inhibit the growth of clinical isolates of drug-resistant Mycobacterium tuberculosis strains as well as M. tuberculosis H37Rv. To investigate the mechanism of ATB107 action in M. tuberculosis, two-dimensional gel electrophoresis coupled with MALDI-TOF-MS analysis (2-DE-MS) was performed to illustrate alterations in the protein expression profile in response to ATB107. Results show that ATB107 affected tryptophan biosynthesis by decreasing the expression of protein encoded by Rv3246c, the transcriptional regulatory protein of MtrA belonging to the MtrA-MtrB two-component regulatory system, in both drug-sensitive and drug-resistant virulent strains. ATB107 might present a stress condition similar to isoniazid (INH) or ethionamide for M. tuberculosis since the altered expression in response to ATB107 of some genes, such as Rv3140, Rv2243, and Rv2428, is consistent with INH or ethionamide treatment. After incubation with ATB107, the expression of 2 proteins encoded by Rv0685 and Rv2624c was down-regulated while that of protein encoded by Rv3140 was up-regulated in all M. tuberculosis strains used in this study. This may be the common response to tryptophan absence; however, relations to ATB107 are unknown and further evaluation is warranted.

Identification of a Methyltransferase Encoded by Gene ste16 and Its Function in Ebosin Biosynthesis of Streptomyces sp. 139: Hong-Guan Xie , Yong-Gang Bao , Li-ping Bai , Jun-Jie Shan , Rong Jiang , Yang Zhang , Lian-Hong Guo , Ren Zhang , Yuan Li; J. Microbiol. 2009;47(2):193-200. Published online May 2, 2009; DOI: https://doi.org/10.1007/s12275-008-0195-y

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Abstract: Streptomyces sp. 139 generates a novel exopolysaccharide (EPS) designated as Ebosin, which exerts an antagonistic effect on IL-1R in vitro and anti-rheumatic arthritis activity in vivo. A ste gene cluster for Ebosin biosynthesis consisting of 27 ORFs was previously identified in our laboratory. In this paper, ste16 was expressed in Escherichia coli BL21 and the recombinant protein was purified, which has the ability to catalyze the transfer of the methyl group from S-adenosylmethionine (AdoMet) to dTDP-4-keto-6-deoxy-D-glucos, which was thus identified as a methyltransferase. In order to determine the function of ste16 in Ebosin biosynthesis, the gene was disrupted with a double crossover via homologous recombination. The monosaccharide composition of EPS-m generated by the mutant strain Streptomyces sp. 139 (ste16-) was found to differ from that of Ebosin. The IL-1R antagonist activity of EPS-m was markedly lower than that of Ebosin. These experimental results have shown that the ste16 gene codes for a methyltransferase which is involved in Ebosin biosynthesis.

cloning of Gene Encoding for Siderophore biosynthesis in Fluorescent Pseudomonas sp.: Koh, Han Cheol , Ha, Sung Cheol , Na, Jung A , Kim, Ho Sang , Yeo, Myeong Gu , Lee, Jung Sup , Kim, Sung Jun , Park, yeal; J. Microbiol. 1995;33(1):28-33.

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Abstract: Pseudomonas sp. strain PY002, isolated from soil, was mutagenized with a transposon Tn5(21). To screening of siderophore biosunthesis defective mutant, 138 kanamycin resistant mutants were tested of growth on MKB medium supplemented with iron chelator(bipydidyl and EDDHA) and in vitro antibiosis. Among 138 mutants, 32 mutants do not excreted a siderophore and lose their antibiotic activity. So, these mutants were designated Flu^-Sid^-. A gene bank of DNA from Pseudomonas sp. strain PY002 was constructed using the broad-host range conjugative cosmid pLAFR3. The recombinant cosmids contained insert DNA averaging 21 kb in length and the frequence of transduction into E. coli HB101 per 1㎍ of insert DNA was 9 × 10³. Nonfluorescent mutants were complemented by mating the gene bank en masse and identifying the 108 fluorescent transconjugants. Restriction enzyme analysis of these complemented transconjugants revealed three different types and they were named pCOM61, pCOM91 and pCOM97. Sizes of their insert DNA were 30kb, 26kb and 28kb, respectively.

Molecular cloning of the arginine biosynthetic genes from corynebacterium glutamicum: Chun, Jae Yeon , Jung, Sam Il , Ko, Soon Young , Park, Mee Young , Kim, Soo Young , Lee, Heung Shickc , Cheon, Choong Ill , Min, Kyung Hee , Lee, Myeong Sok; J. Microbiol. 1996;34(4):355-362.

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Abstract: Complementation cloning of the argC, E, B D, F, and G genes in Corynebacterium glutamicum was done by transforming the genomic DNA library into the corresponding arginine auxotrophs fo Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8kb fragments complementing the E. coli arg B mutant were also able to complement the E. coli argC, E, A, D, and F mutants, indicating the clustered organization of the arginine biosynthetic genes within the cloned DNA fragments. The insert DNA fragments in the recombinant plasmids, named pRB1 AND pRB2, were physically mapped with several restriction enzymes. By further subcloning the entire DNA fragment containing the functions and by complementation analysis, we located the arg genes in the order of ACEBDF on the restriction map. We also determined the DNA nucleotide sequence of the fragment and report here the sequence of the argB gene. When compared to that with the nutant strain, higherh enzyme activity of N-acetylglutamate kinase was detacted in the extract of the mutant carrying the plasmid containing the putative argB gene, indicating that the plasmid contains a functional argB gene. Deduced amino acid sequence of the argB gene shows45%, 38%, and 25% identity to that from Bacillus strearothermophilus, Bacillus substilus, and E. coli respectively. Our long term goal is genetically engineering C. glutamicum which produces more arginine than a wild type strain does.

Expression and Activity of Citrus Phytoene Synthase and [beta]-Carotene Hydroxylase in Escherichia coli: In-Jung Kim , Kyong-Cheol Ko , Tae-Sik Nam , Yu-Wang Kim , Won-Il Chung , Chan-Shick Kim; J. Microbiol. 2003;41(3):212-218.

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Abstract: Citrus phytoene synthase (CitPsy) and [beta]-carotene hydroxylase (CitChx), which are involved in caroteinoid biosynthesis, are distantly related to the corresponding bacterial enzymes from the point of view of amino acid sequence similarity. We investigated these enzyme activities using Pantoea ananatis carotenoid biosynthetic genes and Escherichia coli as a host cell. The genes were cloned into two vector systems controlled by the T7 promoter. SDS-polyacrylamide gel electrophoresis showed that CitPsy and CitChx proteins are normally expressed in E. coli in both soluble and insoluble forms. In vivo complementation using the Pantoea ananatis enzymes and HPLC analysis showed that [beta]-carotene and zeaxanthin were produced in recombinant E. coli, which indicated that the citrus enzymes were functionally expressed in E. coli and assembled into a functional multi-enzyme complex with Pantoea ananatis enzymes. These observed activities well matched the results of other researchers on tomato phytoene synthase and Arabidopsis and pepper [beta]-carotene hydroxylases. Thus, our results suggest that plant carotenoid biosynthetic enzymes can generally complement the bacterial enzymes and could be a means of carotenoid production by molecular breeding and fermentation in bacterial and plant systems.

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