• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.730-738
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• 2016

est19 is a gene from Bacillus sp. K91 that encodes a new esterase. A comparison of the amino acid sequence showed that Est19 has typical Ser-Gly-Asn-His (SGNH) family motifs and could be grouped into the SGNH hydrolase family. The Est19 protein was functionally cloned, and expressed and purified from Escherichia coli BL21(DE3). The enzyme activity was optimal at 60℃ and pH 9.0, and displayed esterase activity towards esters with short-chain acyl esters (C₂-C₆). A structural model of Est19 was constructed using phospholipase A1 from Streptomyces albidoflavus NA297 as a template. The structure showed an α/β-hydrolase fold and indicated the presence of the typical catalytic triad Ser49-Asp227-His230, which were further investigated by site-directed mutagenesis. To the best of our knowledge, Est19 is a new member of the SGNH hydrolase family identified from thermophiles, which may be applicable in the industrial production of semisynthetic β-lactam antibiotics after modification.

• Journal of fish pathology
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• v.30 no.1
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• pp.41-49
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• 2017

Probiotic principles can be applied in aquaculture for the purpose of growth and immunity stimulation, disease prevention and eventually better production performance. This study was to assess effects of combinations of microbes containing two Bacillus sp., plus one Lactobacillus sp. as the basal preparation (BSL-LAB), and additional Nitrosomonas sp. (nitrifying bacteria consortium, NBS) in olive flounder (Paralichthys olivaceus). The effects examined were growth parameters, hematologic parameters, innate immunity and pathogen challenge test. Fish were assigned to 4 treatments as Control (no probiotics), Group A (Bacillius and Lactobacillus to culture water), Group B (Bacillius and Lactobacillus both in water and feed), Group C (same as Group B with additional Nitrosomonas in feed). Fish were allocated to the above 4 groups, each group being composed of triplicate 30 fish, for a 7-week feeding in the laboratory. Positive effects were observed both in growth and pathogen sensitivity with all three probiotic combinations. Such effects were attributed to improved innate immune functions. This result indicates that the tested probiotic microbes are beneficial to olive flounder aquaculture.

• BMB Reports
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• v.32 no.5
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• pp.480-485
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• 1999

Tyrosine phenol-lyase of thermophilic Symbiobacterium sp. SC-1, which is obligately and symbiotically dependent on thermophilic Bacillus sp. SK-1, was purified and characterized. The enzyme is composed of four identical subunits and contains approximately 1 mol of pyridoxal 5'-phosphate (PLP) per mol subunit as a cofactor. The enzyme showed absorption maxima at 330 and 420 nm, and lost this absorption profile by treatment with phenylhydrazine. The apparent dissociation constsnt, $K'_D$, for PLP was determined with the apoenzyme to be about $1.2\;{\mu}M$. The isoelectric point was 4.9. The optimal temperature and pH for the $\alpha,\beta$-elimination of L-tyrosine were found to be $80^{\circ}C$ and pH 8.0, respectively. The substrate specificity of the enzyme was very broad: L-amino acids including L-tyrosine, 3,4-dihydroxyphenyl-L-alanine (L-DOPA), L-cysteine, L-serine, S-methyl-L-cysteine, $\beta$-chloro-L-alanine, and S-(o-nitrophenyl)-L-cysteine all served as substrates. D-Tyrosine and D-serine were also decomposed into pyruvic acid and ammonia at rates of 7% and 31% relative to their corresponding L-enantiomers, respectively. D-Alanine, which was inert as a substrate in a, $\beta$-elimination, was the only D-amino acid racemized by the enzyme. The $K_m$ values for L-tyrosine, L-DOPA, S-(o-nitrophenyl)-L-cysteine, $\beta$-chloro-L-alanine, and S-methyl-L-cysteine were 0.19, 9.9, 0.36, 12, and 5.5 mM, respectively.

• Journal of Applied Biological Chemistry
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• v.54 no.1
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• pp.1-6
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• 2011

${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.

• KSBB Journal
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• v.27 no.2
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• pp.75-85
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• 2012

Although, microbial arsenic mobilization by dissimilatory arsenate-reducing bacteria (DARB) and the practical use to the removal technology of arsenic from contaminated soil are expected, most previous research mainly has been focused on the geochemical circulation of arsenic. Therefore, in this review we summarized the previously reported DARB to grasp the characteristic for bioremediation of arsenic. Evidence of microbial growth on arsenate is presented based on isolate analyses, after which a summary of the physiology of the following arsenate-respiring bacteria is provided: Chrysiogenes arsenatis strain BAL-$1^T$, Sulfurospirillum barnesii, Desulfotomaculum strain Ben-RB, Desulfotomaculum auripigmentum strains OREX-4, GFAJ-1, Bacillus sp., Desulfitobacterium hafniense DCB-$2^T$, strain SES-3, Citrobacter sp. (TSA-1 and NC-1), Sulfurospirillum arsenophilum sp. nov., Shewanella sp., Chrysiogenes arsenatis BAL-$1^T$, Deferribacter desulfuricans. Among the DARB, Citrobacter sp. NC-1 is superior to other dissimilatory arsenate-reducing bacteria with respect to arsenate reduction, particularly at high concentrations as high as 60 mM. A gram-negative anaerobic bacterium, Citrobacter sp. NC-1, which was isolated from arsenic contaminated soil, can grow on glucose as an electron donor and arsenate as an electron acceptor. Strain NC-1 rapidly reduced arsenate at 5 mM to arsenite with concomitant cell growth, indicating that arsenate can act as the terminal electron acceptor for anaerobic respiration (dissimilatory arsenate reduction). To characterize the reductase systems in strain NC-1, arsenate and nitrate reduction activities were investigated with washed-cell suspensions and crude cell extracts from cells grown on arsenate or nitrate. These reductase activities were induced individually by the two electron acceptors. Tungstate, which is a typical inhibitory antagonist of molybdenum containing dissimilatory reductases, strongly inhibited the reduction of arsenate and nitrate in anaerobic growth cultures. These results suggest that strain NC-1 catalyzes the reduction of arsenate and nitrate by distinct terminal reductases containing a molybdenum cofactor. This may be advantageous during bioremediation processes where both contaminants are present. Moreover, a brief explanation of arsenic extraction from a model soil artificially contaminated with As (V) using a novel DARB (Citrobacter sp. NC-1) is given in this article. We conclude with a discussion of the importance of microbial arsenate reduction in the environment. The successful application and use of DARB should facilitate the effective bioremediation of arsenic contaminated sites.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.585-595
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• 2019

Analysis of bacterial communities based on their 16S rDNA sequences revealed the predominance of Proteobacteria (Aeromonas sp., Acinetobacter sp. and Thaueraamino aromatica sp.) and uncultured bacterium in activated sludge from the effluent treatment plant (ETP) of Mother Dairy, Calcutta (India). Each isolate was used for bioremediation of dairy wastewater with simultaneous conversion of nitrogenous pollutants into ammonia. A consortium developed using seven of these isolates and three Bacillus strains from different environmental origins could reduce 93% nitrate with simultaneous production of ammonia (626 ㎍/100 ml) within 20 h in non-aerated, immobilized conditions as compared to 82% nitrate reduction producing 2.4 ㎍/100 ml ammonia in 96 h with extensive aeration in a conventional ETP. The treated ammonia-rich effluent could be used instead of freshwater and fertilizer during cultivation of mung bean with 1.6-fold increase in grain yield. The ETP with the surrounding agricultural land makes this process a zero liquid discharge technology for using the biofertilizer generated. In addition, the process requires minimal energy supporting sustained environmental health. This method is thus proposed as an alternative approach for small-scale dairy ETPs.

• Journal of Life Science
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• v.28 no.6
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• pp.697-707
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• 2018

The aim of this study was to investigate the potential effects of extracts from silkworm (Bombyx mori L.) that was fermented with Bacillus subtilis KACC 91157 at the levels of 1%(v/w), 2.5%(v/w), and 5%(v/w) in alcohol-fed rats. Sprague-Dawley rats were divided into seven groups: normal group (N), alcohol treated group (C), silymarin treated group (PC, positive control), 5% silkworm powder treated group (SP5), 1% Bacillus subtilis-fermented silkworm powder treated group (BSP1), 2.5% BSP treated group (BSP2.5), and 5% BSP treated group (BSP5). The activities of AST, ALT, ALP, and LDH in the serum and the triglyceride concentrations in the liver and serum were increased by alcohol feeding but were reduced in the BSP5 group. In addition, the contents of total lipids, free fatty acids, and total cholesterol were increased in the alcohol-fed group but were reduced in the BSP5 group. The activities of ADH, ALDH and ADH, ALDH protein levels in the liver were increased in the BSP5 group. The TBARS contents in the liver, serum, liver mitochondria, and liver microsomes were slightly decreased in the BSP5 group compared to the C group. The hepatocytes in alcohol-fed rats contained numerous large droplets; however, BSP5 treatment prevented alcohol-induced lipid droplet accumulation in the hepatocytes. Based on these results, extracts from Bacillus subtilis-fermented silkworm (Bombyx mori L.) have significant potential for development into a functional health food that can improve alcoholic fatty liver conditions.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.529.1-529
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• 1986

For isolation of the CMCase gene of the alkalophilic Bacillus sp. strain N-4 to analyze their genetic information for the multicomponents of the cellulase, Bscherichia coli K12 and plasmid DNA pBR322 was used as host-vector system. After the digestion of purified chromosomal DNA and plasmid DNA pBR322 with HindIII, these were ligated. The ligated DND were transformed into Escherichia coli, and recombinant plasmid 107 carried the gene coding for CMCase was constructed. The CMCase produced by Escherichia coli cells containing plasmid DNA pYBC107 was found in the cells as intracellular enzyme and nearly 60% of the total CMCase activity was localized in cellular fraction. Also, the optimum pH for the reaction of CMCase produced by Escherichia coli was appeared at pH .8.0 and the enzyme was stable between pH 7.0 and pH 8.0.

• The Korean Journal of Mycology
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• v.45 no.1
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• pp.43-53
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• 2017

In this study, we analyzed the densities and taxonomic characteristics of various microorganisms that play important roles in Agaricus bisporus culture medium composting, and examined changes in the levels of decomposition-related enzymes secreted by these microorganisms. Various microorganisms such as thermophilic bacteria, actinomycetes, fluorescent Pseudomonas spp., and filamentous bacteria are closely associated with culture medium composts of Agaricus bisporus. The population densities of microorganisms change, and harmful bacteria disappear during thermophilic composting. Psychrobacter sp., Pseudomonas sp., Bacillus sp., and Pseudoxanthomonas sp. accounted for the highest proportion of bacteria in the culture media during outdoor composting, whereas Bacillus sp. and Psychrobacillus sp. were dominant after pasteurization. Cellulose and hemicellulose enzymes of the microorganisms were important at an early stage of rice straw composting and after decomposition of carbon sources, respectively. Microorganisms that secreted these enzymes were present in the second and third turning stage of composting.

• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1029-1039
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• 2024

This study explores beneficial bacteria isolated from the roots and rhizosphere soil of Khao Rai Leum Pua Phetchabun rice plants. A total of 315 bacterial isolates (KK001 to KK315) were obtained. Plant growth-promoting traits (phosphate solubilization and indole-3-acetic acid (IAA) production), and antimicrobial activity against three rice pathogens (Curvularia lunata NUF001, Bipolaris oryzae 2464, and Xanthomonas oryzae pv. oryzae) were assessed. KK074 was the most prolific in IAA production, generating 362.6 ± 28.0 ㎍/ml, and KK007 excelled in tricalcium phosphate solubilization, achieving 714.2 ± 12.1 ㎍/ml. In antimicrobial assays using the dual culture method, KK024 and KK281 exhibited strong inhibitory activity against C. lunata, and KK269 was particularly effective against B. oryzae. In the evaluation of antimicrobial metabolite production, KK281 and KK288 exhibited strong antifungal activities in cell-free supernatants. Given the superior performance of KK281, taxonomically identified as Bacillus sp. KK281, it was investigated further. Lipopeptide extracts from KK281 had significant antimicrobial activity against C. lunata and a minimum inhibitory concentration (MIC) of 3.1 mg/ml against X. oryzae pv. oryzae. LC-ESI-MS/MS analysis revealed the presence of surfactin in the lipopeptide extract. The crude extract was non-cytotoxic to the L-929 cell line at tested concentrations. In conclusion, the in vitro plant growth-promoting and disease-controlling attributes of Bacillus sp. KK281 make it a strong candidate for field evaluation to boost plant growth and manage disease in upland rice.