Journal of Microbiology

Journal Article

The putative sensor histidine kinase VadJ coordinates development and sterigmatocystin production in Aspergillus nidulans: Yanxia Zhao , Mi-Kyung Lee , Jieyin Lim , Heungyun Moon , Hee-Soo Park , Weifa Zheng , Jae-Hyuk Yu; J. Microbiol. 2021;59(8):746-752. Published online July 5, 2021; DOI: https://doi.org/10.1007/s12275-021-1055-2

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The VosA-VelB heterocomplex governs expression of several genes associated with fungal development and secondary metabolism. In this study, we have investigated the functions of one of the VosA-VelB-activated developmental genes vadJ in development and production of the mycotoxin sterigmatocystin in the model fungus Aspergillus nidulans. The vadJ gene is predicted to encode a 957-amino acid length protein containing a highly conserved sensor histidine kinase domain. The deletion of vosA or velB resulted in decreased mRNA levels of vadJ throughout the life cycle, suggesting that VosA and VelB are necessary for proper expression of vadJ. Nullifying vadJ led to highly restricted colony growth, lowered formation of asexual spores, and about two-fold reduction in conidial viability. Conversely, the deletion of vadJ resulted in elevated production of sexual fruiting bodies and sterigmatocystin. These suggest that VadJ is necessary for proper coordination of asexual and sexual development, and sterigmatocystin production. In accordance with this idea, the deletion of vadJ led to elevated mRNA levels of the two key sexual developmental activators esdC and nsdD. In summary, the putative sensor histidine kinase VadJ represses sexual development and sterigmatocystin production, but activates asexual development in A. nidulans.

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Involvement of LaeA and Velvet Proteins in Regulating the Production of Mycotoxins and Other Fungal Secondary Metabolites
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Regulators of the Asexual Life Cycle of Aspergillus nidulans
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Post‐translational modifications drive secondary metabolite biosynthesis in Aspergillus: a review
Kunlong Yang, Jun Tian, Nancy P. Keller
Environmental Microbiology.2022; 24(7): 2857. CrossRef
The Putative C2H2 Transcription Factor VadH Governs Development, Osmotic Stress Response, and Sterigmatocystin Production in Aspergillus nidulans
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Research Support, Non-U.S. Gov't

Diversity of indigenous endophytic bacteria associated with the roots of Chinese cabbage (Brassica campestris L.) cultivars and their antagonism towards pathogens: Md. Azizul Haque , Han Dae Yun , Kye Man Cho; J. Microbiol. 2016;54(5):353-363. Published online April 20, 2016; DOI: https://doi.org/10.1007/s12275-016-5641-7

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Abstract PDF

The study aimed to reveal the diversity of endophytic bacteria in the roots of Chinese cabbage (CC) cultivated in two areas in Korea, namely, Seosang-gun (SS) and Haenam-gun (HN), and also in a transgenic plant (TP) from the laboratory. A total of 653 colonies were isolated from the interior of CC roots, comprising 118, 302, and 233 isolates from SS, HN, and TP samples, respectively. Based on 16S rRNA gene sequence analysis, the isolates belonged to four major phylogenetic groups: high-G+C Gram-positive bacteria (HGC-GPB), low-G+C Gram-positive bacteria (LGC-GPB), Proteobacteria, and Bacteriodetes. The most dominant groups in the roots of the SS, HN, and TP cultivars were LGC-GPB (48.3%), Proteobacteria (50.2%), and HGC-GPB (38.2%), respectively. Importantly, most of the isolates that produced cell-walldegrading enzymes belonged to the genus Bacillus. Bacillus sp. (HNR03, TPR06), Bacillus pumilus (SSR07, HNR11, TPR07), and Bacillus subtilis (TPR03) showed high antagonism against the tested food-borne pathogenic bacteria. In addition, Bacillus sp. (HNR03, TPR06), Bacillus pumilus (SSR07, HNR11, HNR17, TPR11), Microbacterium oxidans (SSR09, TPR04), Bacillus cereus HNR10, Pseudomonas sp. HNR13, and Bacillus subtilis (TPR02, TPR03) showed strong antagonistic activity against the fungi Phythium ultimum, Phytophthora capsici, Fusarium oxysporum, and Rhizoctonia solani. The endophytes isolated from the TP cultivar showed the strongest antagonistic reactions against pathogens. This study is the first report on endophytic bacteria from Chinese cabbage roots.

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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 PDF

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.

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

Characterization of Cellulases of Fungal Endophytes Isolated from Espeletia spp.: Luisa Cabezas , Carolina Calderon , Luis Miguel Medina , Isabela Bahamon , Martha Cardenas , Adriana Jimena Bernal , Andrés Gonzalez , Silvia Restrepo; J. Microbiol. 2012;50(6):1009-1013. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2130-5

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Abstract PDF: Endophytes are microorganisms that asymptomatically invade plant tissues. They can stimulate plant growth and/or provide defense against pathogen attacks through the production of secondary metabolites. Most endophyte species are still unknown, and because they may have several applications, the study of their metabolic capabilities is essential. We characterized 100 endophytes isolated from Espeletia spp., a genus unique to the paramo ecosystem, an extreme environment in the Andean mountain range. We evaluated the cellulolytic potential of these endophytes on the saccharification of the oil palm empty fruit bunch (OPEFB). The total cellulolytic activity was measured for each endophyte on filter paper (FPA). In addition, the specific carboxymethyl cellulase (CMCase), exoglucanase, and β-glucosidase activities were determined. We found four fungi positive for cellulases. Of these fungi, Penicillium glabrum had the highest cellulolytic activity after partial purification, with maximal CMCase, exoglucanase and β-glucosidase enzyme activities of 44.5, 48.3, and 0.45 U/ml, respectively. Our data showed that the bioprospection of fungi and the characterization of their enzymes may facilitate the process of biofuel production.

Journal Article

Screening for Probiotic Properties of Strains Isolated from Feces of Various Human Groups: Sathyaseelan Sathyabama , Rajendran Vijayabharathi , Palanisamy Bruntha devi , Manohar Ranjith kumar , Venkatesan Brindha Priyadarisini; J. Microbiol. 2012;50(4):603-612. Published online July 21, 2012; DOI: https://doi.org/10.1007/s12275-012-2045-1

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Abstract PDF

The present study searched for potential probiotic strains from various human fecal samples. A total of 67 aerobic and 38 anaerobic strains were isolated from 5 different categories of human feces. Systematic procedures were used to evaluate the probiotic properties of the isolated strains. These showed about 75–97% survivability in acidic and bile salt environments. Adhesion to intestinal cell line Caco-2 was also high. The isolates exhibited hydrophobic properties in hexadecane. The culture supernatants of these strains showed antagonistic effects against pathogens. The isolates were resistant to a simulated gastrointestinal environment in vitro. Of the 4 best isolates, MAbB4 (Staphylococcus succinus) and FIdM3 (Enterococcus fecium), were promising candidates for a potential probiotic. S. succinus was found to be a probiotic strain, which is the second such species reported to date in this particular genus. A substantial zone of inhibition was found against Salmonella spp., which adds further support to the suggestion that the probiotic strain could help prevent intestinal infection. This study suggested that the human flora itself is a potential source of probiotics.

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Research Support, N.I.H., Extramural

The Activity of a Small Lytic Peptide PTP-7 on Staphylococcus aureus Biofilms: Riddhi Kharidia , Jun F. Liang; J. Microbiol. 2011;49(4):663-668. Published online September 2, 2011; DOI: https://doi.org/10.1007/s12275-011-1013-5

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Abstract PDF: One of the most important features of bacterial biofilms is their resistance to antibiotics and to the host immune system. In this study, we have found that a small lytic peptide, PTP-7, is very potent to Gram-positive bacteria and is able to kill antibiotic sensitive and resistant Staphylococcus aureus indiscriminately. Further studies have revealed that despite being a cationic peptide, the antibacterial activity of PTP-7 was not affected by the negatively charged extracellular polymeric substance (EPS) of biofilms. PTP-7 could diffuse into the deep layer of S. aureus biofilms to kill bacteria inside biofilms efficiently and effectively. Neither the high concentrations of metal ions nor the acidic pH in biofilms affected the activity of peptide PTP-7. It seems that the unique sequence/structure together with the resistant bacteria killing ability of peptide PTP-7 confers its anti-biofilm activity. This study sheds new light on the treatment of bacterial biofilms, especially various biofilm related infections.

Retracted Publication

Identification and Characterization of a Novel Antibacterial Peptide, Avian β-Defensin 2 from Ducks: Deying Ma , Ruiqin Wang , Wenyan Liao , Zongxi Han , Shengwang Liu; J. Microbiol. 2009;47(5):610-618. Published online October 24, 2009; DOI: https://doi.org/10.1007/s12275-009-0068-z

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Abstract PDF

In this study, a novel avian β-defensin (AvBD) was isolated from duck pancreas. The complete nucleotide sequence of the gene contained an 195 bp open reading frame encoding 64 amino acids. Homology, characterization and comparison of the gene with AvBD from other avian species confirmed that it was duck AvBD2. The mRNA expression of the gene was analyzed in 17 tissues from 21-day-old ducks. AvBD2 was highly expressed in the trachea, crop, heart, bone marrow, and pancreas; moderately expressed in the muscular stomach, small intestine, kidney, spleen, thymus, and bursa of Fabricius; and weakly expressed in skin. We produced and purified recombinant AvBD2 by expressing the gene in Escherichia coli. As expected, the recombinant peptide exhibited strong bactericidal properties against Bacillus cereus, Staphylococcus aureus, and Pasteurella multocida, and weak bactericidal properties against E. coli and Salmonella choleraesuis. In addition, the recombinant protein retained antimicrobial activity against S. aureus under different temperatures (range, -20°C to 100°C) and pH values (range, 3 to 12).

Citations

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Expression of Immune-Related Genes of Ducks Infected with Avian Pathogenic Escherichia coli (APEC)
Rong Li, Ning Li, Jinzhou Zhang, Yao Wang, Jiyuan Liu, Yumei Cai, Tongjie Chai, Liangmeng Wei
Frontiers in Microbiology.2016;[Epub] CrossRef
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Deying Ma, Mingyue Zhang, Kexin Zhang, Xiaoli Liu, Zongxi Han, Yuhao Shao, Shengwang Liu
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Randomized Controlled Trial

Antimicrobial Activity of Enterocins from Enterococcus faecalis SL-5 against Propionibacterium acnes, the Causative Agent in Acne Vulgaris, and Its Therapeutic Effect: Bong Seon Kang , Jae-Gu Seo , Gwa-Su Lee , Jung-Hwa Kim , Sei Yeon Kim , Ye Won Han , Hoon Kang , Hyung Ok Kim , Ji Hwan Rhee , Myung-Jun Chung , Young Min Park; J. Microbiol. 2009;47(1):101-109. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0179-y

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Abstract PDF: A lactic acid bacterial strain was isolated from human fecal specimen and identified as Enterococcus faecalis SL-5. The isolated strain showed antimicrobial activity against Gram-positive pathogens assayed, especially the highest activity against Propionibacterium acnes. The antimicrobial substance was purified and verified as a bacteriocin (named ESL5) of E. faecalis SL-5 by activity-staining using P. acnes as an indicator. N-terminal sequence of ESL5 was determined (MGAIAKLVAK) and sequence analysis revealed that it is almost identical to the some of enterocins including L50A/B of E. faecium L50 and MR10A/B of E. faecalis MRR 10-3. From the sequencing data of L50A/B structural genes, the nucleotide sequence showed 100% identity with that of the MR10A/B structural genes, implying that ESL5 is an equivalent of enterocin MR10. Meanwhile, we also tested the therapeutic effect of anti-P. acnes activity in patients with mild to moderate acne because of its pathogenic role to acne vulgaris. For this purpose, a concentrated powder of CBT SL-5 was prepared using cell-free culture supernatant (CFCS) of E. faecalis SL-5 and included in a lotion for application in the patients. The study showed that CBT SL-5 lotion significantly reduced the inflammatory lesions like pustules compared to the placebo lotion. Therefore our results indicate that the anti-P. acnes activity produced by E. faecalis SL-5 has potential role to the treatment of acne as an alternative to topical antibiotics.

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

Effect of Saliva miltiorrhiza Bunge on Antimicrobial Activity and Resistant Gene Regulation against Methicillin-Resistant Staphylococcus aureus (MRSA): Ji-Won Lee , Young-Ju Ji , Syng-Ook Lee , In-Seon Lee; J. Microbiol. 2007;45(4):350-357.; DOI: https://doi.org/2561 [pii]

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Abstract PDF: This study was conducted in an effort to evaluate the antimicrobial activity and antibiotic-resistant gene regulation from Saliva miltiorrhiza Bunge on methicillin-resistant Staphylococcus aureus (MRSA). A variety of solvent fractions and methanol extracts of S. miltiorrhiza Bunge were tested in order to determine its antimicrobial activities against S. aureus and MRSA. As a result, the hexane fraction of S. miltiorrhiza Bunge evidenced the highest levels of antimicrobial activity against S. aureus and MRSA. The MICs of the hexane fraction against various MRSA specimens were 64<MICs≤128 μg/ml. The hexane fraction evidenced inhibitory effects superior to those of the chloroform fraction. The results showed inhibition zones of hexane (16 mm) and chloroform (14 mm) fractions against MRSA KCCM 40511 at 1,000 μg/disc. The hexane and chloroform fractions inhibited the expression of the resistant genes, mecA, mecR1, and femA in mRNA. Moreover, the results of Western blotting assays indicated that the hexane and chloroform fractions inhibited the expression of the resistant protein, PBP2a. These results reveal that the hexane and chloroform fractions of S. miltiorrhiza Bunge may prove to be a valuable choice for studies targeted toward the development of new antimicrobial agents.

The Antimicrobial Activity of Essential Oil from Dracocephalum foetidum against Pathogenic Microorganisms: Saet Byoul Lee , Kwang Hyun Cha , Su Nam Kim , Shataryn Altantsetseg , Sanduin Shatar , Oidovsambuu Sarangerel , Chu Won Nho; J. Microbiol. 2007;45(1):53-57.; DOI: https://doi.org/2491 [pii]

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Abstract PDF: A number of essential oils from Mongolian aromatic plants are claimed to have antimicrobial activities. The essential oil of Dracocephalum foetidum, a popular essential oil used in Mongolian traditional medicine, was examined for its antimicrobial activity. Eight human pathogenic microorganisms including B. subtilis, S. aureus, M. lutens, E. hirae, S. mutans, E. coli, C. albicans, and S. cerevisiae were examined. The essential oil of Dracocephalum foetidum exhibited strong antimicrobial activity against most of the pathogenic bacteria and yeast strains that were tested; by both the agar diffusion method and the minimum inhibitory concentration (MIC) assay (MIC range was 26-2592 μg/ml). Interestingly, Dracocephalum foetidum even showed antimicrobial activity against methicilin-resistant Staphylococcus aureus (MRSA) strains. We also analyzed the chemical composition of the oil by GC-MS and identified several major components, including n-Mentha-1,8-dien-10-al, limonene, geranial, and neral.

Phylogenic Analysis of Alternaria brassicicola Producing Bioactive Metabolites: Dong-Sun Jung , Yeo-Jung Na , Ki Hyun Ryu; J. Microbiol. 2002;40(4):289-294.

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Abstract PDF: The fungal strain SW-3 having antimicrobial activity was isolated from soil of crucified plants in Pocheon, Kyungki-Do, Korea. Strain SW-3 was identified as Alternaria brassicicola by its morphological characteristics, and confirmed by the analysis of the 18S gene and ITS regions of rDNA. The fungus showed a similarity of 99% with Alternaria brassicicola in the 18S rDNA sequence analysis. A. brassicicola has been reported to produce an antitumor compound, called depudecin. We found that strain SW-3 produced antimicrobial metabolites, in addition to depudecin, during sporulation under different growth conditions. The metabolite of the isolated fungus was found to have strong antifungal activity against Microsporium canis and Trichophyton rubrum, and antibacterial activity against Staphylococcus aureus and Pseudomonas aerogenes. The amount and kind of metabolites produced by the isolate were affected by growth conditions such as nutrients and growth periods.

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