• Journal of Microbiology and Biotechnology
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• 제1권2호
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• pp.102-110
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• 1991

Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1868-1873
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• 2006

A novel microorganism that could degrade high molecular weight gellan was screened and isolated from soil. On gellan plate, the microorganism grew well and completely liquefied the plate. The gellan-degrading microorganism was isolated by pure culture on glucose and nutrient agar medium afterwards. The 16S rDNA sequence analysis and biochemical tests using an API 50CHB/20E kit revealed that the strain belonged to Bacillus sp. The isolate, named as Bacillus sp. YJ-1, showed optimum gellan-degrading activity in 0.5% gellan medium at pH 7.5 and 37$^{\circ}C$. The activity was measured and evaluated by the thiobarbituric acid and thin-layer chromatography method. Mass spectrometry revealed that the major gellan.. depolymerized product was an unsaturated tetrasaccharide consisting of $\Delta$4,5-glucuronic acid-(1$\rightarrow$4 )-$\beta$-D-glucose-(1$\rightarrow$4)- $\alpha$-L-rhamnose-(1$\rightarrow$3)-$\beta$-D-glucose, which is a dehydrated repeating unit of gellan, thus the enzyme was identified as gellan lyase. When the gellan was present in the medium, the gellan-degrading activity was much higher than that in glucose-grown cells. These results indicate that in the presence of gellan, Bacillus sp. YJ-1 is able to metabolize the gellan by inducing gellan-degrading enzymes that can degrade gellan into small molecular weight oligosaccharides, and then the gellan. depolymerized products are taken up by the cells and utilized by intracellular enzymes.

• Journal of Microbiology
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• 제40권2호
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• pp.140-145
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• 2002

The 5.8 kb pMMH1, rolling-circle replication (RCR) plasmid of the wild type soil Bacillus mesentericus was developed into a novel secretion vector system in Bacillus subtilis. The pMMHl turned out to have a replication origin and two open reading frames (ORFs) of the putative γ-GTP and type I signal peptidase (sipP). To characterize the regions necessary for plasmid stability and high copy number, five vectors (pPS, pPP, pEN, pMN, pME) were constructed by disruption or deletion of each region in pMMH1. Like pMMHl all constructed vectors were stable over 100 generations In a non-selective medium. Since pPS was the smallest (2.3 kb)of all, it was selected for the construction of a navel secretion vector, Using the $\alpha$-amylase promoter/signal sequence of B. subtilils the novel plasmid pJSN was constructed. When $\beta$-glucosidase was expressed using pJSN, we found $\beta$-glucosidase activity in the medium. This result strongly suggested that plasmid pJSN can be used for the production of bioactive peptides in B. subtilis.

• Food Science and Biotechnology
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• 제17권2호
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• pp.219-227
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• 2008

Bacillus thuringiensis was isolated as a pathogen of the sotto disease of silkmoth larvae about a hundred years ago. Since then, this bacterium has attracted attentions of not only insect pathologists but also many other scientists who are interested in its strong and specific insecticidal activity. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as a landmark discovery of par asp orin, a cancer cell-specific cytotoxin produced by B. thuringiensis. In this review, we describe examination of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using a surface plasmon resonance-based biosensor, identification and characterization of parasporin-4, the latest parasporin produced by the B. thuringiensis A1470 strain, and an effective method for preparing the parasporin-4 from inclusion bodies expressed in the recombinant Escherichia coli cells.

• 한국임상수의학회지
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• 제18권1호
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• pp.18-21
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• 2001

In this study, we assessed the efficacy of a novel B. amyloliquefacience DS11 phytase (DS11 phytase) and that of a commercial Aspergillus ficcum phytase (AF phytase) through their bioavailabilities of phytin-posphorus and -calcium in the diet using cannulated pigs. For the purpose of evaluating the efficacy of the phytases in pigs, we determined phosphorous concentrations from serum and feces, in addition to ingesta obtained from the cannula at the terminal ileum. As results, phosphorus concentration was lower in feces from DS11 group and BASF group by 17% and 10%, and higher in serum from the respective groups by 34% and 20%, as compared to the control group. Both phytases are evaluated to enhance phosphorus availability to the great extent. Calcium concentration of feces were lower in DS11 group and BASF group by 31% and 10%, than that in the control. Calcium concentration of serum was higher in DS11 phytase group by 4% but lower in AF phyase group by 3%, then that in the control group.

• Journal of Microbiology and Biotechnology
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• 제24권10호
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• pp.1405-1412
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• 2014

Lysozyme is a protein found in egg white, tears, saliva, and other secretions. As a marketable natural alternative to preservatives, lysozyme can act as a natural antibiotic. In this study, we have isolated Bacillus licheniformis TIB320 from soil, which contains a lysozyme gene with various features. We have cloned and expressed the lysozyme in E. coli. The antimicrobial activity of the lysozyme showed that it had a broad antimicrobial spectrum against several standard strains. The lysozyme could maintain efficient activities in a pH range between 3 and 9 and from $20^{\circ}C$ to $60^{\circ}C$, respectively. The lysozyme was resistant to pepsin and trypsin to some extent at $40^{\circ}C$. Production of the lysozyme was optimized by using various expression strategies in B. subtilis WB800. The lysozyme from B. licheniformis TIB320 will be promising as a food or feed additive.

• 환경생물
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• 제36권2호
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• pp.199-205
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• 2018

Petroleum energy is the major source of the world energy market, and its massive usage, and the corresponding extreme environmental pollution, imposes a serious threat on the ecological cycles. By screening oil-contaminated soil, we isolated, identified, and characterized a novel strain that represents a considerable diesel-degrading potentiality; the Bacillus aryabhattai DA2 strain is registered in the NCBI with the accession number MG571630, and it possesses an efficient tributyrin-degrading capacity. The optimal condition for diesel degradation by DA2 strain was observed at pH between 7-8 and at the temperature of $30^{\circ}C$. The strain is resistant to salt as well as the antibiotics like ampicillin and streptomycin. These results indicate B. aryabhattai is one of the potential candidates for the remediation of the diesel-contaminated sites.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.798-803
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• 2001

Peptide synthetases produced from various microorganisms are multifunctional enzyme complexes and their substrates are recognized and activated by adenylation domains. To identify the substrate specificity of the peptide synthetase isolated from Bacillus subtilis 713, known to produce an antifungal peptide, two adenylation domains containing the minimal functional portion were expressed and purified. ATP-ppi exchange experiments and kinetic studies revealed that the two adenylation enzymes had a substrate specificity to $_{L}-lysine{\;}and{\;}_{L}-tryptophan$, respectively. In addition, based on a signature sequence comparison, the substrate of the third domain was predicted to be L-glutamic acid. These results suggest that this peptide synthetase is novel because there has been no previous report on a peptide synthetase that uses $_{L}-lysine,{\;}_{L}-tryptophan,{\;}and{\;}_{L}-glutamic$ acid as substrates in that order.

• 미생물학회지
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• 제34권3호
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• pp.87-90
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• 1998

우리 나라의 전통 대두발효식품인 된장으로부터 혈전용해능을 가진 미생물을 분리하였다. 그 중 5종류의 균주를 Bergey's manual of systematic bacteriology에 의거하여 동정한 결과 Bacillus sp. 균주로 밝혀졌다. 분리된 Bacillus 속 미생물을 효소 유도배지에서 배양한 결과 B. amyloliquefaciens는 2.46 plasmin unit/ml, B pantothenticus는 3.82 plasmin unit/ml 의 혈전용해능을 가지고 있었고, B. subtilis는 4.94 plasmin unti/ml의 높은 혈전용해효소 생산능을 보여주었다. 분리 균주에 의하여 생산된 세포외 단백질을 SDS-PAGE와 reverse fibrin zymogram 활성측정법에 의해 확인한 결과 각 균주별로 서로 다른 혈전용해효소가 생산되었음을 확인하였다.

• 한국미생물·생명공학회지
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• 제34권1호
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• pp.52-57
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• 2006

새로운 고지혈증 예방 제품 개발을 위한 기초 자료를 얻고자 우선 전통 된장으로부터 HMG-CoA reductase 저해 물질을 강력하게 세포외로 생산하는 D-3 세균을 최종 분리 하였다. D-3 균의 미생물학적 특성을 조사한 결과 그람양성의 간균으로 운동성이 있었고 통성 혐기성이였다. 이들 특성과 16S rRNA 염기서열의 분석 결과 등을 종합하여 Bergey's manual로 동정한 결과 Bacillus cereus D-3로 동정되었다. HMG-CoA reductase 저해물질 생산 최적 조건을 조사한 결과 Bacillus cereus D-3를 glucose 2%, corn steep liquor $0.6%,\;K_{2}HPO_4\;0.04%,\;KH_{2}PO_4\;0.05%$를 함유한 Glucose-CSL 배지에 접종하여 $30^{\circ}C$에서 36시간 배양하여 얻은 상등액의 HMC-CoA reductase 저해활성이 39.4% 저해물질을 가장 많이 생성 하였다.