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Advances in functional analysis of the microbiome: Integrating metabolic modeling, metabolite prediction, and pathway inference with Next-Generation Sequencing data: Sungwon Jung; J. Microbiol. 2025;63(1):e.2411006. Published online January 24, 2025; DOI: https://doi.org/10.71150/jm.2411006

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Abstract PDF: This review explores current advancements in microbiome functional analysis enabled by next-generation sequencing technologies, which have transformed our understanding of microbial communities from mere taxonomic composition to their functional potential. We examine approaches that move beyond species identification to characterize microbial activities, interactions, and their roles in host health and disease. Genome-scale metabolic models allow for in-depth simulations of metabolic networks, enabling researchers to predict microbial metabolism, growth, and interspecies interactions in diverse environments. Additionally, computational methods for predicting metabolite profiles offer indirect insights into microbial metabolic outputs, which is crucial for identifying biomarkers and potential therapeutic targets. Functional pathway analysis tools further reveal microbial contributions to metabolic pathways, highlighting alterations in response to environmental changes and disease states. Together, these methods offer a powerful framework for understanding the complex metabolic interactions within microbial communities and their impact on host physiology. While significant progress has been made, challenges remain in the accuracy of predictive models and the completeness of reference databases, which limit the applicability of these methods in under-characterized ecosystems. The integration of these computational tools with multi-omic data holds promise for personalized approaches in precision medicine, allowing for targeted interventions that modulate the microbiome to improve health outcomes. This review highlights recent advances in microbiome functional analysis, providing a roadmap for future research and translational applications in human health and environmental microbiology.

Research Article

Lactic acid bacteria from Ethiopian traditional beverage, Tella: technological and metabolic profiles for industrial application: Gashaw Assefa Yehuala, Jaein Choe, Nurelegne Tefera Shibeshi, Kumsa Delessa, Asnake Desalegn, Mi-Kyung Park; J. Microbiol. 2025;63(1):e.2409008. Published online December 20, 2024; DOI: https://doi.org/10.71150/jm.2409008

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Abstract PDF: Tella is a traditional beverage widely accepted by consumers, despite the lack of product consistency owing to its reliance on natural fermentation. This study aimed to identify potential industrial lactic acid bacteria (LAB) starter cultures based on their technological properties. Seven LAB strains isolated from Tella were characterized for their carbohydrate utilization, salt content, temperature, and acid tolerances, growth and acidification rates, and metabolite profiles. Most strains efficiently utilized various carbohydrates, with Lactiplantibacillus plantarum TDM41 showing exceptional versatility. The strains exhibited similar growth characteristics. Principal component analysis of stress tolerance properties revealed that L. plantarum TDM41, Pediococcus pentosaceus TAA01, and Leuconostoc mesenteroides TDB22 exhibited superior tolerance ability. Strong acidification properties were detected in the L. plantarum TDM41, P. pentosaceus TAA01, and Leuconostoc mesenteroides TDB22 strains after 24 h incubation at 30°C. L. plantarum TDM41 displayed the fastest acidification rate throughout the analysis period. All LAB strains produced significant amounts of diverse organic acids, including lactic acid, citric acid, acetic acid, malic acid, and succinic acid, with lactic acid being the primary acid produced by each strain. Overall, strains L. plantarum TDM41 and P. pentosaceus TAA01 prove to be potential candidates for Tella industrial starter cultures and similar cereal products owing to their robust technological properties.

Journal Articles

Hydroxychloroquine an Antimalarial Drug, Exhibits Potent Antifungal Efficacy Against Candida albicans Through Multitargeting: Sargun Tushar Basrani, Tanjila Chandsaheb Gavandi, Shivani Balasaheb Patil, Nandkumar Subhash Kadam, Dhairyasheel Vasantrao Yadav, Sayali Ashok Chougule, Sankunny Mohan Karuppayil, Ashwini Khanderao Jadhav; J. Microbiol. 2024;62(5):381-391. Published online April 8, 2024; DOI: https://doi.org/10.1007/s12275-024-00111-6

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Abstract: Candida albicans is the primary etiological agent associated with candidiasis in humans. Unrestricted growth of C. albicans can progress to systemic infections in the worst situation. This study investigates the antifungal activity of Hydroxychloroquine (HCQ) and mode of action against C. albicans. HCQ inhibited the planktonic growth and yeast to hyphal form morphogenesis of C. albicans significantly at 0.5 mg/ml concentration. The minimum inhibitory concentrations (MIC(50)) of HCQ for C. albicans adhesion and biofilm formation on the polystyrene surface was at 2 mg/ml and 4 mg/ml respectively. Various methods, such as scanning electron microscopy, exploration of the ergosterol biosynthesis pathway, cell cycle analysis, and assessment of S oxygen species (ROS) generation, were employed to investigate HCQ exerting its antifungal effects. HCQ was observed to reduce ergosterol levels in the cell membranes of C. albicans in a dose-dependent manner. Furthermore, HCQ treatment caused a substantial arrest of the C. albicans cell cycle at the G0/G1 phase, which impeded normal cell growth. Gene expression analysis revealed upregulation of SOD2, SOD1, and CAT1 genes after HCQ treatment, while genes like HWP1, RAS1, TEC1, and CDC 35 were downregulated. The study also assessed the in vivo efficacy of HCQ in a mice model, revealing a reduction in the pathogenicity of C. albicans after HCQ treatment. These results indicate that HCQ holds for the development of novel antifungal therapies.

Non-mitochondrial aconitase regulates the expression of iron-uptake genes by controlling the RNA turnover process in fission yeast: Soo-Yeon Cho , Soo-Jin Jung , Kyoung-Dong Kim , Jung-Hye Roe; J. Microbiol. 2021;59(12):1075-1082. Published online October 26, 2021; DOI: https://doi.org/10.1007/s12275-021-1438-4

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Abstract

Aconitase, a highly conserved protein across all domains of life, functions in converting citrate to isocitrate in the tricarboxylic acid cycle. Cytosolic aconitase is also known to act as an iron regulatory protein in mammals, binding to the RNA hairpin structures known as iron-responsive elements within the untranslated regions of specific RNAs. Aconitase-2 (Aco2) in fission yeast is a fusion protein consisting of an aconitase and a mitochondrial ribosomal protein, bL21, residing not only in mitochondria but also in cytosol and the nucleus. To investigate the role of Aco2 in the nucleus and cytoplasm of fission yeast, we analyzed the transcriptome of aco2ΔN mutant that is deleted of nuclear localization signal (NLS). RNA sequencing revealed that the aco2ΔN mutation caused increase in mRNAs encoding iron uptake transporters, such as Str1, Str3, and Shu1. The half-lives of mRNAs for these genes were found to be significantly longer in the aco2ΔN mutant than the wild-type strain, suggesting the role of Aco2 in mRNA turnover. The three conserved cysteines required for the catalytic activity of aconitase were not necessary for this role. The UV cross-linking RNA immunoprecipitation analysis revealed that Aco2 directly bound to the mRNAs of iron uptake transporters. Aco2-mediated degradation of iron-uptake mRNAs appears to utilize exoribonuclease pathway that involves Rrp6 as evidenced by genetic interactions. These results reveal a novel role of non-mitochondrial aconitase protein in the mRNA turnover in fission yeast to fine-tune iron homeostasis, independent of regulation by transcriptional repressor Fep1.

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Iron-mediated post-transcriptional regulation in Toxoplasma gondii
Megan A. Sloan, Adam Scott, Dana Aghabi, Lucia Mrvova, Clare R. Harding, Dominique Soldati-Favre
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The Key Enzymes of Carbon Metabolism and the Glutathione Antioxidant System Protect Yarrowia lipolytica Yeast Against pH-Induced Stress
Tatyana I. Rakhmanova, Natalia N. Gessler, Elena P. Isakova, Olga I. Klein, Yulia I. Deryabina, Tatyana N. Popova
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The intricate link between iron, mitochondria and azoles in Candida species
Wouter Van Genechten, Rudy Vergauwen, Patrick Van Dijck
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Non-Mitochondrial Aconitase-2 Mediates the Transcription of Nuclear-Encoded Electron Transport Chain Genes in Fission Yeast
Ho-Jung Kim, Soo-Yeon Cho, Soo-Jin Jung, Yong-Jun Cho, Jung-Hye Roe, Kyoung-Dong Kim
Journal of Microbiology.2024; 62(8): 639. CrossRef
Kinetic and Regulatory Properties of Yarrowia lipolytica Aconitate Hydratase as a Model-Indicator of Cell Redox State under pH Stress
Tatyana I. Rakhmanova, Varvara Yu. Sekova, Natalya N. Gessler, Elena P. Isakova, Yulia I. Deryabina, Tatyana N. Popova, Yevgeniya I. Shurubor, Boris F. Krasnikov
International Journal of Molecular Sciences.2023; 24(8): 7670. CrossRef

Root-associated bacteria influencing mycelial growth of Tricholoma matsutake (pine mushroom): Seung-Yoon Oh , Young Woon Lim; J. Microbiol. 2018;56(6):399-407. Published online June 1, 2018; DOI: https://doi.org/10.1007/s12275-018-7491-y

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Abstract

Tricholoma matsutake is an ectomycorrhizal fungus usually associated with Pinus densiflora in South Korea. Fruiting bodies (mushrooms) of T. matsutake are economically important due to their attractive aroma; yet, T. matsutake is uncultivatable and its habitat is rapidly being eradicated due to global climate change. Root-associated bacteria can influence the growth of ectomycorrhizal fungi that co-exist in the host rhizosphere and distinctive bacterial communities are associated with T. matsutake. In this study, we investigated how these bacterial communities affect T. matsutake growth by isolating bacteria from the roots of P. densiflora colonized by ectomycorrhizae of T. matsutake and co-culturing rootassociated bacteria with T. matsutake isolates. Thirteen species of bacteria (27 isolates) were found in pine roots, all belonging to the orders Bacillales or Burkholderiales. Two species in the genus Paenibacillus promoted the growth of T. matsutake in glucose poor conditions, likely using soluble metabolites. In contrast, other bacteria suppressed the growth of T. matsutake using both soluble and volatile metabolites. Antifungal activity was more frequent in glucose poor conditions. In general, pine rhizospheres harbored many bacteria that had a negative impact on T. matsutake growth and the few Paenibacillus species that promoted T. matsutake growth. Paenibacillus species, therefore, may represent a promising resource toward successful cultivation of T. matsutake.

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Haiyun Zi, Yun Wang, Yangwenke Liao, Shuikuan Bei, Jinchi Zhang, Xiaogang Li
Applied Soil Ecology.2025; 206: 105833. CrossRef
Cultivable Root-Symbiotic Bacteria of a Pioneer Ericaceous Dwarf Shrub Colonizing Volcanic Deposits and Their Potential to Promote host Fitness
Alejandra Fuentes-Quiroz, Héctor Herrera, Roxana Alvarado, Cristiane Sagredo-Saez, Maria Isabel-Mujica, Martin Vohník, Eleonora Rolli
Journal of Soil Science and Plant Nutrition.2024; 24(2): 3355. CrossRef
Commodity chain as a negotiated process: Empirical analysis of benefit allocation, governance, and powers of upstream and downstream actors in matsutake mushroom trade in Shangri-La, Yunnan Province
Xinyang Li, Jinlong Liu, Ryo Kohsaka
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Min-Jeong Kang, Eun-Kyung Bae, Eung-Jun Park, Kang-Hyeon Ka, Mi-Ra Son, Ki-Tae Kim, Jei-Wan Lee, Jason E. Stajich
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Afshana, Zafar A. Reshi, Manzoor A. Shah, Rayees A. Malik, Irfan Rashid
International Microbiology.2023; 26(4): 1053. CrossRef
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Yixin Zhou, Zhichao Shi, Qiliang Pang, Xiufeng Liang, Hongtao Li, Xin Sui, Chongwei Li, Fuqiang Song
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Bioactive properties of streptomyces may affect the dominance of Tricholoma matsutake in shiro
Lu-Min Vaario, Shumpei Asamizu, Tytti Sarjala, Norihisa Matsushita, Hiroyasu Onaka, Yan Xia, Hiroyuki Kurokochi, Shin-Ichi Morinaga, Jian Huang, Shijie Zhang, Chunlan Lian
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Farzad Aslani, Leho Tedersoo, Sergei Põlme, Oliver Knox, Mohammad Bahram
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Tingye Feng, Sang Eun Jeong, Jin Ju Lim, Seogang Hyun, Che Ok Jeon
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PLOS ONE.2018; 13(11): e0205900. CrossRef

Proteome analysis reveals global response to deletion of mrflbA in Monascus ruber: Qingqing Yan , Zhouwei Zhang , Yishan Yang , Fusheng Chen , Yanchun Shao; J. Microbiol. 2018;56(4):255-263. Published online February 28, 2018; DOI: https://doi.org/10.1007/s12275-018-7425-8

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Abstract

Monascus spp. are commonly used for a wide variety of applications in the food and pharmaceutical industries. In previous studies, the knock-out of mrflbA (a putative regulator of the G protein α subunit) in M. ruber led to autolysis of the mycelia, decreased pigmentation and lowered mycotoxin production. Therefore, we aimed to obtain a comprehensive overview of the underlying mechanism of mrflbA deletion at the proteome level. A two-dimensional gel electrophoresis analysis of mycelial proteins indicated that the abundance of 178 proteins was altered in the ΔmrflbA strain, 33 of which were identified with high confidence. The identified proteins are involved in a range of activities, including carbohydrate and amino acid metabolism, hyphal development and the oxidative stress response, protein modification, and the regulation of cell signaling. Consistent with these findings, the activity of antioxidative enzymes and chitinase was elevated in the supernatant of the ΔmrflbA strain. Furthermore, deletion of mrflbA resulted in the transcriptional reduction of secondary metabolites (pigment and mycotoxin). In short, the mutant phenotypes induced by the deletion of mrflbA were consistent with changes in the expression levels of associated proteins, providing direct evidence of the regulatory functions mediated by mrflbA in M. ruber.

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Histone deacetylase MrHos3 negatively regulates the production of citrinin and pigments in Monascus ruber
Qianrui Liu, Yunfan Zheng, Baixue Liu, Fufang Tang, Yanchun Shao
Journal of Basic Microbiology.2023; 63(10): 1128. CrossRef
Histone deacetylase MrRpd3 plays a major regulational role in the mycotoxin production of Monascus ruber
Yunfan Zheng, Yueyan Huang, Zejing Mao, Yanchun Shao
Food Control.2022; 132: 108457. CrossRef
Characterization of key upstream asexual developmental regulators in Monascus ruber M7
Lili Jia, Yuyun Huang, Jae-Hyuk Yu, Marc Stadler, Yanchun Shao, Wanping Chen, Fusheng Chen
Food Bioscience.2022; 50: 102153. CrossRef
Quantitative Proteomics Analysis by Sequential Window Acquisition of All Theoretical Mass Spectra–Mass Spectrometry Reveals Inhibition Mechanism of Pigments and Citrinin Production of Monascus Response to High Ammonium Chloride Concentration
Bo Zhou, Yifan Ma, Yuan Tian, Jingbo Li, Haiyan Zhong
Journal of Agricultural and Food Chemistry.2020; 68(3): 808. CrossRef

A computationally simplistic poly-phasic approach to explore microbial communities from the Yucatan aquifer as a potential sources of novel natural products: Marfil-Santana Miguel David , O’Connor-Sánchez Aileen , Ramírez-Prado Jorge Humberto , De los Santos-Briones Cesar , López- Aguiar , Lluvia Korynthia , Rojas-Herrera Rafael , Lago-Lestón Asunción , Prieto-Davó Alejandra; J. Microbiol. 2016;54(11):774-781. Published online October 29, 2016; DOI: https://doi.org/10.1007/s12275-016-6092-x

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Abstract

The need for new antibiotics has sparked a search for the microbes that might potentially produce them. Current sequencing technologies allow us to explore the biotechnological potential of microbial communities in diverse environments without the need for cultivation, benefitting natural product discovery in diverse ways. A relatively recent method to search for the possible production of novel compounds includes studying the diverse genes belonging to polyketide synthase pathways (PKS), as these complex enzymes are an important source of novel therapeutics. In order to explore the biotechnological potential of the microbial community from the largest underground aquifer in the world located in the Yucatan, we used a polyphasic approach in which a simple, non-computationally intensive method was coupled with direct amplification of environmental DNA to assess the diversity and novelty of PKS type I ketosynthase (KS) domains. Our results suggest that the bioinformatic method proposed can indeed be used to assess the novelty of KS enzymes; nevertheless, this in silico study did not identify some of the KS diversity due to primer bias and stringency criteria outlined by the metagenomics pipeline. Therefore, additionally implementing a method involving the direct cloning of KS domains enhanced our results. Compared to other freshwater environments, the aquifer was characterized by considerably less diversity in relation to known ketosynthase domains; however, the metagenome included a family of KS type I domains phylogenetically related, but not identical, to those found in the curamycin pathway, as well as an outstanding number of thiolases. Over all, this first look into the microbial community found in this large Yucatan aquifer and other fresh water free living microbial communities highlights the potential of these previously overlooked environments as a source of novel natural products.

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Biosynthetic potential of the sediment microbial subcommunities of an unexplored karst ecosystem and its ecological implications
Pablo Suárez‐Moo, Alejandra Prieto‐Davó
MicrobiologyOpen.2024;[Epub] CrossRef
Changes in the sediment microbial community structure of coastal and inland sinkholes of a karst ecosystem from the Yucatan peninsula
Pablo Suárez-Moo, Claudia A. Remes-Rodríguez, Norma A. Márquez-Velázquez, Luisa I. Falcón, José Q. García-Maldonado, Alejandra Prieto-Davó
Scientific Reports.2022;[Epub] CrossRef
Insights into the Chemical Diversity of Selected Fungi from the Tza Itzá Cenote of the Yucatan Peninsula
Carlos A. Fajardo-Hernández, Firoz Shah Tuglak Khan, Laura Flores-Bocanegra, Alejandra Prieto-Davó, Baojie Wan, Rui Ma, Mallique Qader, Rodrigo Villanueva-Silva, Anahí Martínez-Cárdenas, Marian A. López-Lobato, Shabnam Hematian, Scott G. Franzblau, Huzefa
ACS Omega.2022; 7(14): 12171. CrossRef

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

Characterization of NpgA, a 4'-phosphopantetheinyl transferase of Aspergillus nidulans, and evidence of its involvement in fungal growth and formation of conidia and cleistothecia for development: Jung-Mi Kim , Ha-Yeon Song , Hyo-Jin Choi , Kum-Kang So , Dae-Hyuk Kim , Keon-Sang Chae , Dong-Min Han , Kwang-Yeop Jahng; J. Microbiol. 2015;53(1):21-31. Published online January 4, 2015; DOI: https://doi.org/10.1007/s12275-015-4657-8

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Abstract

The null pigmentation mutant (npgA1) in Aspergillus nidulans
results
in a phenotype with colorless organs, decreased branching growth, delayed of asexual spore development, and aberrant cell wall structure. The npgA gene was isolated from A. nidulans to investigate these pleiomorphic phenomena of npgA1 mutant. Sequencing analysis of the complementing gene indicated that it contained a 4􍿁-phosphopantetheinyl transferase (PPTase) superfamily domain. Enzymatic assay of the PPTase, encoded by the npgA gene, was implemented in vivo and in vitro. Loss-of-function of LYS5, which encoded a PPTase in Saccharomyces cerevisiae, was functionally complemented by NpgA, and Escherichia coli-derived NpgA revealed phosphopantetheinylation activity with the elaboration of 3􍿁5􍿁-ADP. Deletion of the npgA gene caused perfectly a lethal phenotype and the absence of asexual/sexual sporulation and secondary metabolites such as pigments in A. nidulans. However, a cross feeding effect with A. nidulans wild type allowed recovery from deletion defects, and phased-culture filtrate from the wild type were used to verify that the npgA gene was essential for formation of metabolites needed for development as well as growth. In addition, forced expression of npgA promoted the formation of conidia and cleistothecia as well as growth. These results indicate that the npgA gene is involved in the phosphopantetheinylation required for primary biological processes such as growth, asexual/sexual development, and the synthesis of secondary metabolites in A. nidulans.

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Liamocins biosynthesis, its regulation in Aureobasidium spp., and their bioactivities
Xin-Xin Kang, Shu-Lei Jia, Xin Wei, Mei Zhang, Guang-Lei Liu, Zhong Hu, Zhe Chi, Zhen-Ming Chi
Critical Reviews in Biotechnology.2022; 42(1): 93. CrossRef
Shimalactone Biosynthesis Involves Spontaneous Double Bicyclo‐Ring Formation with 8π‐6π Electrocyclization
Isao Fujii, Makoto Hashimoto, Kaori Konishi, Akiko Unezawa, Haruka Sakuraba, Kenta Suzuki, Harue Tsushima, Miho Iwasaki, Satsuki Yoshida, Akane Kudo, Rina Fujita, Aika Hichiwa, Koharu Saito, Takashi Asano, Jun Ishikawa, Daigo Wakana, Yukihiro Goda, Ayumi
Angewandte Chemie.2020; 132(22): 8542. CrossRef
Shimalactone Biosynthesis Involves Spontaneous Double Bicyclo‐Ring Formation with 8π‐6π Electrocyclization
Isao Fujii, Makoto Hashimoto, Kaori Konishi, Akiko Unezawa, Haruka Sakuraba, Kenta Suzuki, Harue Tsushima, Miho Iwasaki, Satsuki Yoshida, Akane Kudo, Rina Fujita, Aika Hichiwa, Koharu Saito, Takashi Asano, Jun Ishikawa, Daigo Wakana, Yukihiro Goda, Ayumi
Angewandte Chemie International Edition.2020; 59(22): 8464. CrossRef
Genomic analysis of a riboflavin-overproducing Ashbya gossypii mutant isolated by disparity mutagenesis
Tatsuya Kato, Junya Azegami, Ami Yokomori, Hideo Dohra, Hesham A. El Enshasy, Enoch Y. Park
BMC Genomics.2020;[Epub] CrossRef
Genetic evidences for the core biosynthesis pathway, regulation, transport and secretion of liamocins in yeast-like fungal cells
Si-Jia Xue, Guang-Lei Liu, Zhe Chi, Zhi-Chao Gao, Zhong Hu, Zhen-Ming Chi
Biochemical Journal.2020; 477(5): 887. CrossRef
A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene,npgA, inAspergillus nidulans
Ha-Yeon Song, Dahye Choi, Dong-Min Han, Dae-Hyuk Kim, Jung-Mi Kim
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Both a PKS and a PPTase are involved in melanin biosynthesis and regulation of Aureobasidium melanogenum XJ5-1 isolated from the Taklimakan desert
Hong Jiang, Guang-Lei Liu, Zhe Chi, Jian-Ming Wang, Ly-Ly Zhang, Zhen-Ming Chi
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Kum-Kang So, Yun-Jo Chung, Jung-Mi Kim, Beom-Tae Kim, Seung-Moon Park, Dae-Hyuk Kim
Molecules and Cells.2015; 38(12): 1105. CrossRef

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: 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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