• Title/Summary/Keyword: microbial strain

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Molecular Cloning and Heterologous Expression of an Acid-Stable Endoxylanase Gene from Penicillium oxalicum in Trichoderma reesei

  • Wang, Juan;Mai, Guoqin;Liu, Gang;Yu, Shaowen
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.251-259
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    • 2013
  • An endoxylanase gene (PoxynA) that belongs to the glycoside hydrolase (GH) family 11 was cloned from a xylanolytic strain, Penicillium oxalicum B3-11(2). PoxynA was overexpressed in Trichoderma reesei QM9414 by using a constitutive strong promoter of the encoding pyruvate decarboxylase (pdc). The high extracellular xylanase activities in the fermentation liquid of the transformants were maintained 29~35-fold higher compared with the wild strain. The recombinant POXYNA was purified to homogeneity, and its characters were analyzed. Its optimal temperature and pH value were $50^{\circ}C$ and 5.0, respectively. The enzyme was stable at a pH range of 2.0 to 7.0. Using beechwood as the substrate, POXYNA had a high specific activity of $1,856{\pm}53.5$ IU/mg. In the presence of metal ions, such as $Cu^{2+}$, and $Mg^{2+}$, the activity of the enzyme increased. However, strong inhibition of the enzyme activity was observed in the presence of $Mn^{2+}$ and $Fe^{2+}$. The recombinant POXYNA hydrolyzed birchwood xylan, beechwood xylan, and oat spelt xylan to produce short-chain xylooligosaccharides, xylopentaose, xylotriose, and xylobiose as the main products. This is the first report on the expression properties of a recombinant endoxylanase gene from Penicillium oxalicum. The properties of this endoxylanase make it promising for applications in the food and feed industries.

Positive Regulation of Pyoluteorin Biosynthesis in Pseudomonas sp. M18 by Quorum-Sensing Regulator VqsR

  • Huang, Xianqing;Zhang, Xuehong;Xu, Yuquan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.828-836
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    • 2008
  • The biocontrol rhizobacterium Pseudomonas sp. M18 can produce two kinds of antibiotics, namely pyoluteorin (Plt) and phenazine-1-carboxylic acid (PCA), and is antagonistic against a number of soilborne phytopathogens. In this study, a luxR-type quorum-sensing regulatory gene, vqsR, was identified and characterized immediately downstream of the Plt gene cluster in strain MI8. A vqsR-inactivated mutant led to a significant decrease in the production of Plt and its biosynthetic gene expression. However, this was restored when introducing the vqsR gene by cloning into the plasmid pME6032 in trans. The vqsR mutation did not exert any obvious influence on the production of PCA and its biosynthetic gene expression and the production of N-acylhomoserine lactones (C4 and C8-HSLs) and their biosynthetic gene rhlI expression. Accordingly, these results introduce VqsR as a regulator of Plt production in Pseudomonas spp., and suggest that the regulatory mechanism of vqsR in strain M18 is distinct from that in P. aeruginosa. In addition, it was demonstrated that vqsR mutation did not have any obvious impact on the expression of Plt-specific ABC transporters and other secondary metabolic global regulators, including GacA, RpoS, and RsmA.

Isolation of Indole-3-acetic acid (IAA) producing Arthrobacter sp. and plant growth promotion effect (Indole-3-acetic acid (IAA) 생성 Arthrobacter sp.의 분리 및 식물 생육촉진 효과)

  • Da Som Kim;Ho-Young Shin;Song-Ih Han
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.6
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    • pp.831-838
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    • 2022
  • An auxin-producing bacteria, KSD16, KSD33, and KSD36 were isolated from agricultural soil. The strain KSD16, KSD33, and KSD36 was classified as a strain of Arthrobacter sp. based on phylogenetic analysis of 16S rRNA gene. The isolated KDS16, KDS33, and KSD36 was confirmed to produce indole-3-acetic acid (IAA), which is one of the auxin hormones. When the concentration of IAA was assessed the maximum concentration of IAA, 206.62 mg L-1, was detected from the culture broth incubated in R2A medium containing 0.1% L-tryptophan for 48 h at 28 ℃. To study the effect of IAA producing bacteria on germination rate, seeds of Mung bean were prepared for each treatment. KSD16, KSD33, and KSD36 showed significant increase in root length and number of adventitious roots than the controls. To investigate the growth-promoting effects on the crops, Arthrobacter species were placed in water cultures and seed pots of mung beans. In consequence, the seed germination of mung beans was 73.4% higher than the control.

Comparison of microbial molecular diagnosis efficiency within unstable template metagenomic DNA samples between qRT-PCR and chip-based digital PCR platforms

  • Dongwan Kim;Junhyeon Jeon;Minseo Kim;Jinuk Jeong;Young Mok Heo;Dong-Geol Lee;Dong Keon Yon;Kyudong Han
    • Genomics & Informatics
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    • v.21 no.4
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    • pp.52.1-52.10
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    • 2023
  • Accurate and efficient microbial diagnosis is crucial for effective molecular diagnostics, especially in the field of human healthcare. The gold standard equipment widely employed for detecting specific microorganisms in molecular diagnosis is quantitative real-time polymerase chain reaction (qRT-PCR). However, its limitations in low metagenomic DNA yield samples necessitate exploring alternative approaches. Digital PCR, by quantifying the number of copies of the target sequence, provides absolute quantification results for the bacterial strain. In this study, we compared the diagnostic efficiency of qRT-PCR and digital PCR in detecting a particular bacterial strain (Staphylococcus aureus), focusing on skin-derived DNA samples. Experimentally, specific primer for S. aureus were designed at transcription elongation factor (greA) gene and the target amplicon were cloned and sequenced to validate efficiency of specificity to the greA gene of S. aureus. To quantify the absolute amount of microorganisms present on the skin, the variable region 5 (V5) of the 16S rRNA gene was used, and primers for S. aureus identification were used to relative their amount in the subject's skin. The findings demonstrate the absolute convenience and efficiency of digital PCR in microbial diagnostics. We suggest that the high sensitivity and precise quantification provided by digital PCR could be a promising tool for detecting specific microorganisms, especially in skin-derived DNA samples with low metagenomic DNA yields, and that further research and implementation is needed to improve medical practice and diagnosis.

Concanamycin B, Active substance Against Phytophthora capsici Produced by Streptomyces neyagawaensis 38D10 Strain (Streptomyces neyagawaensis 38D10 균주가 생산하는 concanamycin B의 항고추역병 활성)

  • Kim, Chang-Jin;Lee, In-Kyoung;Yun, Bong-Sik;Yoo, Ick-Dong
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.322-328
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    • 1993
  • During the screening of antifungal compounds from microbial secondary metabolites to control phytophthora blight of red pepper caused by Phytophthora capsici, a soil isolate, strain 38D10 was selected. Based on taxonomic studies, this strain was identified as Streptomyces neyagawaensis. The antifungal compound was purified from culture broth by HP-20 column chromatography, ethyl acetate extraction, silica gel column chromatography, HPLC and identified as concanamycin B by UV. $^1H$-NMR, $^{13}C$-NMR, SIMS analysis. Concanamycin B has strong antifungal activity against some phytopathogenic fungi but not antivacterial activity and preventive value were 50% and 100% at 125ppm and 250ppm in pot assay.

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The effect of environmental factors affecting to the growth of Rhizobium japonicum (Rhizobium japonicum의 생장 및 poly$\beta$hydroxybutyric acid(PHB)의 축적에 미치는 환경요인의 영향)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.20 no.4
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    • pp.195-200
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    • 1982
  • Rhizobium japonicum was isolated from the nodule of soybean root grown at the reclaimed tidal land in Kang-Wha island. The effect of pH and salt concentration to the viability of the isolated strain were examined in relationship between microbial growth and accumulation of PHB. Optimal pH value for the good viability of the isolated strain was 7.0 and also, at 5.0 and 6.0 viability was favorable to large extent, but 9.0 was unfavorable. Examined the effect of salt concentration treated two times as of the salinity in the reclaimed tidal land, viability of the isolated strain showed about 30 to 40%. And also in treatment with NaCl(40g/l) whatever the pH value adopted, viability was mostly less than 10%. The amount of accumulated PHB was relatively high at low pH value(5-6) and at high salt concentrration, respectively.

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Current Status and Applications of Adaptive Laboratory Evolution in Industrial Microorganisms

  • Lee, SuRin;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.793-803
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    • 2020
  • Adaptive laboratory evolution (ALE) is an evolutionary engineering approach in artificial conditions that improves organisms through the imitation of natural evolution. Due to the development of multi-level omics technologies in recent decades, ALE can be performed for various purposes at the laboratory level. This review delineates the basics of the experimental design of ALE based on several ALE studies of industrial microbial strains and updates current strategies combined with progressed metabolic engineering, in silico modeling and automation to maximize the evolution efficiency. Moreover, the review sheds light on the applicability of ALE as a strain development approach that complies with non-recombinant preferences in various food industries. Overall, recent progress in the utilization of ALE for strain development leading to successful industrialization is discussed.

Production of Cellulosic Ethanol in Saccharomyces cerevisiae Heterologous Expressing Clostridium thermocellum Endoglucanase and Saccharomycopsis fibuligera β-glucosidase Genes

  • Jeon, Eugene;Hyeon, Jeong-eun;Suh, Dong Jin;Suh, Young-Woong;Kim, Seoung Wook;Song, Kwang Ho;Han, Sung Ok
    • Molecules and Cells
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    • v.28 no.4
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    • pp.369-373
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    • 2009
  • Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and ${\beta}$-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an ${\alpha}$-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and ${\beta}$-glucosidase was able to produce ethanol from ${\beta}$-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst.

Isolation of Alkalophilic Bacillus sp. KJ-133 Producing Cyclomaltodextrinase and Its Enzyme Production (Cyclomaltodextrinase를 생산하는 Alkalophilic Bacillus sp. KJ-133의 분리와 효소생산 조건)

  • 정혜진;권호정
    • Microbiology and Biotechnology Letters
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    • v.28 no.4
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    • pp.219-222
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    • 2000
  • To produce and utilize microbial cyclomaltodextrinase being industrially useful, we isolated an alkalophilic Bacillus strain from soil which was capable of degrading cyclodextrins. The newly isolated strain was aerobic, gram-positive, spore-forming, motile, rod shape(0.2~0.4$\times$1.4~4.4 $\mu\textrm{m}$), and 35.8 mol% of DNA base composition. Based on its morphological, phisiological, and biochemical properties, it was identified as alkalophilic Bacillus sp. KJ-133 and cultivated well in the ranges of $30~40^{\circ}C$ and pH 8.0~9.0 . The cyclomaltodextrinase of the strain showed maximal production after 48h of cultivation at $37^{\circ}C$, and the activity was inhibited by Ag2+, Hg2+, Cu2+, and p-chloromercuribenzoate.

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Effect of inoculation of Lactobacillus plantarum isolated from swine feces on fermentation characteristics of hulless barley

  • Jeong, Yong Dae;Lee, Jung Jae;Seol, Kuk-Hwan;Kim, Doo Wan;Min, Ye Jin;Yu, Dong Jo;Cho, Kyu Ho;Kim, Young Hwa
    • Korean Journal of Agricultural Science
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    • v.44 no.4
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    • pp.558-565
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    • 2017
  • This study was conducted to determine the effect of inoculation of microorganism isolated from pig feces on nutrient contents of fermented hulless barley. The microbial flora in feces of a total of four crossbred piglets ($Landrace{\times}Yorkshire{\times}Duroc$) was analyzed by 16s rRNA sequencing. The most abundant strain was then selected for fermentation of hulless barley. Lactobacillus plantarum (L. plantarum) was dominant (64.56%) in intestinal microbial flora in the pig feces. The selected candidate strain showed significantly higher survival rate at pH 7 than at pH 2.5 and 3.0 (p < 0.05). Incubated culture containing the candidate strain showed an increased growth rate with lower pH levels after 7.5 h incubation compared to initial incubation period (p < 0.05). When compared with commercial multiple probiotics which were used as control, the selected strain showed faster growth rate at 5 h post-incubation (p < 0.05). During the fermentation period, neither inoculated nor non-inoculated control hulless barley showed any change in pH value. Crude fat, fiber and ash contents were lower (p < 0.05) in hulless barley inoculated by the selected strain compared to control. However, moisture, energy, NDF and ADF were not affected by the inoculation of strain or fermentation period. Lactic acid was increased and acetic acid was decreased in the hulless barley inoculated with the selected strain during the fermentation period (p < 0.05). Taken together, our results suggest that L. plantarum derived from the pigs could be utilized as a new microorganism for manufacturing fermented feed stuffs.