• Title/Summary/Keyword: Secondary metabolite biosynthesis

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Characterization of Pseudomonas sp. NIBR-H-19, an Antimicrobial Secondary Metabolite Producer Isolated from the Gut of Korean Native Sea Roach, Ligia exotica

  • Sungmin Hwang;Jun Hyeok Yang;Ho Seok Sim;Sung Ho Choi;Byounghee Lee;Woo Young Bang;Ki Hwan Moon
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1416-1426
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    • 2022
  • The need to discover new types of antimicrobial agents has grown since the emergence of antibiotic-resistant pathogens that threaten human health. The world's oceans, comprising complex niches of biodiversity, are a promising environment from which to extract new antibiotics-like compounds. In this study, we newly isolated Pseudomonas sp. NIBR-H-19 from the gut of the sea roach Ligia exotica and present both phenotypes and genomic information consisting of 6,184,379 bp in a single chromosome possessing a total of 5,644 protein-coding genes. Genomic analysis of the isolated species revealed that numerous genes involved in antimicrobial secondary metabolites are predicted throughout the whole genome. Moreover, our analysis showed that among twenty-five pathogenic bacteria, the growth of three pathogens, including Staphylococcus aureus, Streptococcus hominis and Rhodococcus equi, was significantly inhibited by the culture of Pseudomonas sp. NIBR-H-19. The characterization of marine microorganisms with biochemical assays and genomics tools will help uncover the biosynthesis and action mechanism of antimicrobial metabolites for development as antagonistic probiotics against fish pathogens in an aquatic culture system.

Enhanced Salt Stress Tolerance in Transgenic Potato Plants Expressing IbMYB1, a Sweet Potato Transcription Factor

  • Cheng, Yu-Jie;Kim, Myoung-Duck;Deng, Xi-Ping;Kwak, Sang-Soo;Chen, Wei
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1737-1746
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    • 2013
  • IbMYB1, a transcription factor (TF) for R2R3-type MYB TFs, is a key regulator of anthocyanin biosynthesis during storage of sweet potatoes. Anthocyanins provide important antioxidants of nutritional value to humans, and also protect plants from oxidative stress. This study aimed to increase transgenic potatoes' (Solanum tuberosum cv. LongShu No.3) tolerance to environmental stress and enhance their nutritional value. Transgenic potato plants expressing IbMYB1 genes under the control of an oxidative stress-inducible peroxidase (SWPA2) promoter (referred to as SM plants) were successfully generated through Agrobacterium-mediated transformation. Two representative transgenic SM5 and SM12 lines were evaluated for enhanced tolerance to salinity, UV-B rays, and drought conditions. Following treatment of 100 mM NaCl, seedlings of SM5 and SM12 lines showed less root damage and more shoot growth than control lines expressing only an empty vector. Transgenic potato plants in pots treated with 400 mM NaCl showed high amounts of secondary metabolites, including phenols, anthocyanins, and flavonoids, compared with control plants. After treatment of 400 mM NaCl, transgenic potato plants also showed high DDPH radical scavenging activity and high PS II photochemical efficiency compared with the control line. Furthermore, following treatment of NaCl, UV-B, and drought stress, the expression levels of IbMYB1 and several structural genes in the flavonoid biosynthesis such as CHS, DFR, and ANS in transgenic plants were found to be correlated with plant phenotype. The results suggest that enhanced IbMYB1 expression affects secondary metabolism, which leads to improved tolerance ability in transgenic potatoes.

Improved mevinolic acid (MA) production by the immobilized cells, and the establishment of on-line measurement system for fermentation parameters using vent gas analyzer

  • Song, Seong-Gi;Kim, Gyeong-Hui;Kim, Myeong-Jin;Lee, Sang-Jong;Jang, Yong-Geun;Jeong, Yeon-Ho;Jeong, Yong-Seop;Jeon, Gye-Taek
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.223-227
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    • 2003
  • Mevinolic acid (MA), a secondary metabolite produced by a filamentous fungus Aspergillus terreus, is acidic form of lovastatin which has been identified as a powerful cholesterol-lowering agent in humans. When immobilized cell culture was performed, MA production was about 5.3-fold higher than the parallel suspended cell culture. Although the immobilized cells proliferated slowly during exponential in comparison with the suspended cells, oxygen uptake rate and oxygen mass transfer coefficient of the immobilized cell culture were about 1.3- and 2.5- fold higher respectively than those of the parallel suspended cell culture. From these results, it was concluded that MA biosynthesis was closely dependent on the cell growth rate, morphology and oxygen availability.

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Draft Genome Sequence of the Neodothiora populina-Like Yeast Strain JAF-11, Which Produces the Biosurfactant myo-Inositol Lipids

  • Jeong-Seon Kim;Parthiban Subramanian;Seunghwan Kim;Jun Heo;Bong-Sik Yun;Yiseul Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.328-331
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    • 2023
  • Genomic information of biotechnologically and industrially important microorganisms provides the basis for understanding their metabolic potential. Here, we report the draft genome sequence of the Neodothiora populina-like yeast strain JAF-11 capable of producing biosurfactant myo-inositol lipids. The draft genome contained genes associated with secondary metabolite biosynthesis, including transport and metabolism of lipids, which are a major component of fungal surfactants. Identification of myo-inositol and acyl chain synthesis genes in the draft genome corresponded to the specific biosurfactant produced by strain JAF-11. Further experimental studies could help to elucidate the genes responsible for the production of biosurfactant by strain JAF-11.

Genomics and LC-MS Reveal Diverse Active Secondary Metabolites in Bacillus amyloliquefaciens WS-8

  • Liu, Hongwei;Wang, Yana;Yang, Qingxia;Zhao, Wenya;Cui, Liting;Wang, Buqing;Zhang, Liping;Cheng, Huicai;Song, Shuishan;Zhang, Liping
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.417-426
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    • 2020
  • Bacillus amyloliquefaciens is an important plant disease-preventing and growth-promoting microorganism. B. amyloliquefaciens WS-8 can stimulate plant growth and has strong antifungal properties. In this study, we sequenced the complete genome of B. amyloliquefaciens WS-8 by Pacific Biosciences RSII (PacBio) Single Molecule Real-Time (SMRT) sequencing. The genome consists of one chromosome (3,929,787 bp) and no additional plasmids. The main bacteriostatic substances were determined by genome, transcriptome, and mass spectrometry data. We thereby laid a theoretical foundation for the utilization of the strain. By genomic analysis, we identified 19 putative biosynthetic gene clusters for secondary metabolites, most of which are potentially involved in the biosynthesis of numerous bioactive metabolites, including difficidin, fengycin, and surfactin. Furthermore, a potential class II lanthipeptide biosynthetic gene cluster and genes that are involved in auxin biosynthesis were found. Through the analysis of transcriptome data, we found that the key bacteriostatic genes, as predicted in the genome, exhibited different levels of mRNA expression. Through metabolite isolation, purification, and exposure experiments, we found that a variety of metabolites of WS-8 exert an inhibitory effect on the necrotrophic fungus Botrytis cinerea, which causes gray mold; by mass spectrometry, we found that the main substances are mainly iturins and fengycins. Therefore, this strain has the potential to be utilized as an antifungal agent in agriculture.

The Protein Kinase A Pathway Regulates Zearalenone Production by Modulating Alternative ZEB2 Transcription

  • Park, Ae Ran;Fu, Minmin;Shin, Ji Young;Son, Hokyoung;Lee, Yin-Won
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.967-974
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    • 2016
  • Zearalenone (ZEA) is an estrogenic mycotoxin that is produced by several Fusarium species, including Fusarium graminearum. One of the ZEA biosynthetic genes, ZEB2, encodes two isoforms of Zeb2 by alternative transcription, forming an activator (Zeb2L-Zeb2L homooligomer) and an inhibitor (Zeb2L-Zeb2S heterodimer) that directly regulate the ZEA biosynthetic genes in F. graminearum. Cyclic AMP-dependent protein kinase A (PKA) signaling regulates secondary metabolic processes in several filamentous fungi. In this study, we investigated the effects of the PKA signaling pathway on ZEA biosynthesis. Through functional analyses of PKA catalytic and regulatory subunits (CPKs and PKR), we found that the PKA pathway negatively regulates ZEA production. Genetic and biochemical evidence further demonstrated that the PKA pathway specifically represses ZEB2L transcription and also takes part in posttranscriptional regulation of ZEB2L during ZEA production. Our findings reveal the intriguing mechanism that the PKA pathway regulates secondary metabolite production by reprograming alternative transcription.

Mass Production of Gain-of-Function Mutants of Hairy Roots in Catharanthus roseus (일일초에서의 기능획득 돌연변이 모상근의 대량생산)

  • Ko, Suk-Min;Chung, Hwa-Jee;Lee, Hyo-Yeon
    • Korean Journal of Plant Resources
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    • v.24 no.5
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    • pp.514-520
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    • 2011
  • This study describes conditions for the mass production of mutant hairy root lines by co-cultivation with A. rhizogenes harboring the activation tagging vector pHC7. Various sources of explants were subjected to genetic transformation with A. rhizogenes to determine optimum conditions and cultivar for the highest frequency of hairy root formation on explants. Hairy root formation also were investigated in transformed hairy roots grown in various culture media. Finally, a total of approximately 2,500 lines of hairy root mutants were produced in this study. A managing system for metabolomics in hairy root lines also were established. These hairy root lines will be useful to determine functions of genes relating biosynthesis pathway of secondary metabolites.

Functional Expression of Proteomics-guided AfsR2-dependent Genes in Avermectin-producing Streptomyces avermitilis (Avermectin을 생산하는 Streptomyces avermitilis에서의 Proteomics-guided AfsR2-dependent 유전자의 발현)

  • Kim Myung-Gun;Park Hyun-Joo;Im Jong-Hyuk;Kim Eung-Soo
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.211-215
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    • 2006
  • AfsR2 is a global regulatory protein involved in the stimulation of secondary metabolite biosynthesis in various Streptomyces species including avermectin-producing S. avermitilis. Among several AfsR2-dependent genes identified from the comparative proteomics, the polyribonucleotide nucleotidyltransferase (PNP) and the glyceraldehyde-3-phosphate dehydrogenase (GPD) genes were previously proposed to regulate the actinorhodin production in S. lividans upon afsR2 over-expression positively and negatively, respectively. To show the biological significance of the PNP and GPD genes in the S. avermitilis strains, these two genes were functionally expressed in both the wild-type and the avermectin-overproducing mutant strains. The PNP gene expression stimulated secondary metabolite production in the wild-type S. avermitilis ATCC31267, but not in the avermectin-overproducing S. avermitilis ATCC31780. Interestingly, the GDP gene expression stimulated secondary metabolite production by 4-fold in the wild-type S. avermitilis ATCC31267 and by 2.5-fold in the avermectin-overproducing S. avermitilis ATCC31780, respectively. These results suggest that the biological significance of the afsR2-dependent PNP and GPD gene expressions on antibiotic biosynthetic regulation could be significantly different depending on Streptomyces species.

Transcriptome analysis of a medicinal plant, Pistacia chinensis

  • Choi, Ki-Young;Park, Duck Hwan;Seong, Eun-Soo;Lee, Sang Woo;Hang, Jin;Yi, Li Wan;Kim, Jong-Hwa;Na, Jong-Kuk
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.274-281
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    • 2019
  • Pistacia chinensis Bunge has not only been used as a medicinal plant to treat various illnesses but its young shoots and leaves have also been used as vegetables. In addition, P. chinensis is used as a rootstock for Pistacia vera (pistachio). Here, the transcriptome of P. chinensis was sequenced to enrich genetic resources and identify secondary metabolite biosynthetic pathways using Illumina RNA-seq methods. De novo assembly resulted in 18,524 unigenes with an average length of 873 bp from 19 million RNA-seq reads. A Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation tool assigned KO (KEGG orthology) numbers to 6,553 (36.2%) unigenes, among which 4,061 unigenes were mapped into 391 different metabolic pathways. For terpenoid backbone and carotenoid biosynthesis pathways, 44 and 22 unigenes encode enzymes corresponding to 30 and 16 entries, respectively. Twenty-two unigenes encode proteins for 16 entries of the carotenoid biosynthesis pathway. As for the phenylpropanoid and flavonoid biosynthesis pathways, 63 and 24 unigenes were homologous to 17 and 14 entry proteins, respectively. Mining of simple sequence repeat identified 2,599 simple sequence repeats from P. chinensis unigenes. The results of the present study provide a valuable resource for in-depth studies on comparative and functional genomics to unravel the underlying mechanisms of the medicinal properties of Pistacia L.