• Title/Summary/Keyword: Bacillus sp. K91

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Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

  • Maeng, Chang-Jae;Kim, Hyung-Kwoun;Park, Sun-Yang;Koo, Bon-Tag;Oh, Tae-Kwang;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.85-91
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    • 2001
  • To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

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Studies on the Production of Thermostable Amylase. Part 1. Optimal Culture Conditions and Purification of Enzyme. (내열성 Amylase의 생산에 관한 연구 (제1보) 최적배양조건과 효소의 정제)

  • 오두환;이강표;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.9 no.2
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    • pp.91-97
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    • 1981
  • A thermophilic soil isolate Bacillus sp. Y-127 was selected for the production of thermostable amylase. The strain was used for the enzyme production and the thermostable amylase was characterized. The optimum cultural conditions for the enzyme production were 6$0^{\circ}C$ at pH 7.0 for 32 hours using a mineral medium containing 2% soluble starch and 0.2% yeast extract. The extra-cellular enzyme was purified about 123-folds with about 6% recovery. The purified enzyme was stable at pH between 4.0 and 7.0, and temperature up to 6$0^{\circ}C$.

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Comparative Characterization of Xylanases from Two Bacillus Strains (두 종류 Bacillus속 균주의 Xylanases 특성 비교)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.370-375
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    • 2016
  • Two xylanase genes were cloned into Escherichia coli from Bacillus sp. YB-1401 and B. amyloliquefaciens YB-1402, which had been isolated as mannanase producer from home-made doenjang, respectively, and their nucleotide sequences were determined. Both xylanase genes consisted of 642 nucleotides, encoding polypeptides of 213 amino acid residues. The deduced amino acid sequences of the YB-1401 and YB-1402 xylanase, designated Xyn1401 and Xyn1402, differed from each other by single amino acid residue, Asn for Xyn1401 and Lys for Xyn1402, corresponding to amino acid position of 127. Their amino acid sequences were highly homologous to those of xylanases belonging to the glycosyl hydrolase family 11. The 28 amino acid stretch in the N-terminus of both enzymes was predicted as signal peptide by SignalP4.1 server. Both xylanases were localized at the level of 91−94% in culture filtrate of the recombinant E. coli cells, suggesting they were secreted efficiently in E. coli cells. The optimal reaction conditions were 50℃ and pH 6.0 for Xyn1401, and 55℃ and pH 6.5 for Xyn1402, respectively, indicating one amino acid difference from each other affected pH and temperature profiles of their activities. In addition, their thermostabilities were somewhat different from each other.

Comparative Analysis of Pathogenic Organisms in Cockroaches from Different Community Settings in Edo State, Nigeria

  • Isaac, Clement;Orue, Philip Ogbeide;Iyamu, Mercy Itohan;Ehiaghe, Joy Imuetiyan;Isaac, Osesojie
    • Parasites, Hosts and Diseases
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    • v.52 no.2
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    • pp.177-181
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    • 2014
  • Cockroaches are abundant in Nigeria and are seen to harbour an array of pathogens. Environmental and sanitary conditions associated with demographic/socio-economic settings of an area could contribute to the prevalence of disease pathogens in cockroaches. A total of 246 cockroaches (Periplaneta americana) in urban (Benin, n=91), semi-urban (Ekpoma, n=75) and rural (Emuhi, n=70) settings in Edo State, Nigeria were collected within and around households. The external body surfaces and alimentary canal of these cockroaches were screened for bacterial, fungal, and parasitological infections. Bacillus sp. and Escherichia coli were the most common bacteria in cockroaches. However, Enterococcus faecalis could not be isolated in cockroaches trapped from Ekpoma and Emuhi. Aspergillus niger was the most prevalent fungus in Benin and Ekpoma, while Mucor sp. was predominant in Emuhi. Parasitological investigations revealed the preponderance of Ascaris lumbricoides in Benin and Emuhi, while Trichuris trichura was the most predominant in Ekpoma. The prevalence and burden of infection in cockroaches is likely to be a reflection of the sanitary conditions of these areas. Also, cockroaches in these areas making incursions in homes may increase the risk of human infections with these disease agents.

Establishment of $F_o-value$ Criterion for Canned Smoked-Oyster In Cottonseed Oil (훈제 굴 통조림의 가열살균기준 설정에 관한 연구)

  • HAN Bong-Ho;LEE Chang-Kook;IM Chi-Won;YU Hong-Sik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.3
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    • pp.347-353
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    • 1995
  • [ $F_o-value$ ] of canned smoked-oyster in cottonseed oil (SOCO) were measured using a microcomputer aided $F_o-value$ measuring system, and the microbiological safety of the canned SOCO was evaluated to optimize the energy consumption. Most of the microorganisms in raw oyster were saprophytes. No microorganisms were detected from the canned SOCO which was pretreated by conventional procedure and sterilized at $110^{\circ}C$ with $F_o-value$ of 5.92min and over. The most heat resistant microflora isolated from the raw oyster was Bacillus sp.. D-value at $121.1^{\circ}C$ and z-value of spores of Bacillus sp. in the SOCO homogenate were 4.10min and $10,91^{\circ}C$, respectively. After 120 days storage at $50^{\circ}C$, no growth of microorganisms was recognized from the canned SOCO with $F_o-value$of 5.92min.

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Red-Colored Phenomena of Ginseng(Panax ginseng C. A. Meyer) Root and Soil Environment (인삼근 적변현상과 근권 토양환경)

  • 양덕조;김용해
    • Journal of Ginseng Research
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    • v.21 no.2
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    • pp.91-97
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    • 1997
  • In order to elucidate the mechanism of red-colored phenomena(RCP) in ginseng(Panax ginseng C.A. Meyer), distribution of inorganic elements of ginseng root and its surrounding soil, and microflora in the soil were investigated. Red brown colored-substances were accumulated in the cell wall of epidermis at early stage of red-colored ginseng (RCG). Cell wall of the late stage of RCG was disordered and microorganisms were shown in the disordered cell wall. Al, Si and Fe contents among inorpanic elements in the epidermis of RCG were higher at two or three times than that of healthy ginseng. On the other hand, K content was higher at three times in healthy ginseng than that of RCG. Especially, Fe content was higher at three times in lateral roots of RCG than that of healthy ginseng. Total 21 strains of microorganisms were isolated on the 523 medium from surface soil, surrounding soil of both healthy and RCG, and RCG. Six strains of microorganisms among them were resistant to 2 mM Fe. Two species in Bacillus app. and Lactobacillus app. , and one species in Micrococcus sp. and Npisseria sp. respectively were identified. It seemed that RCP was closely related with the distribution and uptake of inorganic elements, was also correlated Fe-independent metabolism of microorganisms.

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Quality Characteristics of Kochujang Meju Prepared with Aspergillus Species and Bacillus subtilis

  • Oh, Byoung-Hak;Kim, Yong-Suk;Jeong, Pyeong-Hwa;Shin, Dong-Hwa
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.549-554
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    • 2006
  • To standardize a manufacturing method and improve the quality of traditional kochujang, eight-types of meju with different shapes (brick, grain) were prepared using Aspergillus oryzae (A.o) or Aspergillus sojae (A.s) alone or in combination with Bacillus subtilis (B.s). The physicochemical characteristics and enzyme activities of the various meju were compared during fermentation for 12 days at $28^{\circ}C$. The moisture content of both the brick- and grain-shaped meju were gradually decreased from an initial content of 50.47 to 54.89% to a content of 12.91 to 16.25% on day 12 of fermentation. The neutral protease activities of the brick-shaped meju ranged from $1.19{\pm}0.12$ to $1.25{\pm}0.28\;unit/mL$, and were similar for all treatments. The ${\alpha}$-amylase activities in A.s+B.s treatment of brick-shaped and grain-shaped meju were the highest, $11{\pm}0.6$ and $9{\pm}0.7\;unit/mL$, respectively. The ${\beta}$-amylase activities ranged from $1.53{\pm}0.01$ to $1.56{\pm}0.02\;unit/mL$, and were similar for all treatments. The amino type nitrogen content of A.o+B.s brick-shaped meju was the highest, $0.39{\pm}0.03%$. We confirmed that the brick-shaped meju prepared with A. oryzae and B. subtilis could be used to prepare traditional kochujang to improve the quality of the product.

Fermentation characteristics of mulberry (Cudrania tricuspidata) fruits produced using microbes isolated from traditional fermented food, and development of fermented soybean food (전통장류로부터 분리한 발효미생물을 이용한 꾸지뽕 열매 발효물의 특성 및 장류제품 개발)

  • Lee, Eun-Sil;Jo, Seung-Wha;Yim, Eun-Jung;Kim, Yun-Sun;Park, Hae-Suk;Kim, Myung-Kon;Cho, Sung-Ho
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.866-877
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    • 2014
  • The aim of this study was to develop a new functional traditional fermented soybean food using Cudrania tricuspidata fruits and fermentation microbes isolated from traditional fermented food. Aspergillus oryzae koji, Lactobacillus sp., and Bacillus sp. were used for the selection of a suitable microbe for the fermentation of Cudrania tricuspidata fruits, and as a result, Bacillus licheniformis SCDB 1234 was selected. SCDB 1234 enhanced the concentration of kaempferol in the Cudrania tricuspidata fruits from 9.54 to $217.04{\mu}g/g$ (about 22 times). The DPPH radical scavenging activity of the fermented materials was similar to that of BHA and BHT (92~99 ppm). The tyrosinase inhibitory activity was high with arbutin (95 ppm) and kojic acid (90 ppm). Doenjang-added fermentation materials of the Cudrania tricuspidata fruits were developed, and the organic acid, reducing sugar, and free amino acid of the developed Doenjang were analyzed. The pancreatic lipase inhibitory (PLI) activity and ${\alpha}$-glucosidase inhibitory (AGI) activity of the fermentation materials of the Cudrania tricuspidata fruits and the developed Doenjang were investigated, and it was found that after fermentation, the PLI and AGI activities of the fermentation materials of the Cudrania tricuspidata fruits were higher than those before fermentation, and that the AGI activity of the developed Doenjang after aging ($91.25{\pm}0.04%$) was higher than that before aging ($84.89{\pm}0.08%$).

A Study on the Measurement of the Pollution Level of Bacteria and Disinfection of Table and IP Cassette (촬영 테이블과 IP Cassette의 세균 오염도 측정 및 소독에 관한 연구)

  • Bae, Seok-Hwan;Lee, Moo-Sik;Lim, Chang-Seon;Kim, Gha-Jung
    • Journal of radiological science and technology
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    • v.31 no.3
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    • pp.229-237
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    • 2008
  • Purpose : For the number of microbes and the pollution level of bacteriology of IP Cassette and Table by laboratories, after identifying the bacteria before and after using alcohol and tissue including disinfectant and statistically testing, this research was intended to provide the basic data for the prevention and the right disinfection guideline for infection management of hospitals in diagnostic radiology. Subject and Method : The subject of this research was the general room of diagnostic radiology of a university hospital in Daejeon City. The research was conducted from Apr 5 to Apr 12, 2007. The number of microbes and the pollution level of bacteriology of IP Cassette and Table by laboratories were tested before and after using alcohol and tissue for disinfection including disinfectant. In order to collect specimens exactly, they were collected with the nurse who specialized in infection management of the hospital, and statistical processing was done with SPSS V13.0. To compare the results before and after using alcohol and tissue, T-test was implemented, and post-hoc test was conducted. Results : Bacteria were detected in 19 cases of 24 subjects(79.2%), however, they were not detected in 5 cases(20.8%). 7kinds of bacteria were detected as isolated bacteria, of which Methicillin Resistant coagulase-negative Staphylococci(MRCNS) were detected in 15 cases(62.5%), which was most, Methicillin Resistant Staphylococcus Aureus(MRSA) in 6 cases(16.7%), Enterococcus Faecium(EFM) in 5 cases(20.8%), Acinetobacter baumannii(ABA) in 2 cases(8.3%), and Bacillus sp, Coagulase-negative Staphylococci(CNS), and Enterococcus sp(ENT) in 1 case, respectively(4.2%). In all bacteria except ABA, Gram positive bacillia were detected in 30 cases(97%), and Gram negative bacilli were detected only in 1 case(3%). As for the kinds of bacteria and the number of groups before and after using 70% Alcohol by Groups, when the bacteria were identified after disinfecting IP Cassette and Table with 70% Alcohol, all the bacteria became extinct in both IP Cassette and Table Group(100%). As for the kinds and number of bacteria before and after using Tissue Cleaner by Groups, the bacteria in Tissue Cleaner Group became completely extinct only in 10 Groups(71.2%), and in 4 Groups, there was much decrease in bacteria, however, they were still detected. The extinction rate of all the bacteria was 91.5%. That is, though the other bacteria became extinct(100%), that of MRCNS bacteria was lowest(83.6%), followed by MRSA(95%). Conclusion : As a result of comparing the mean of the bacteria which were detected before and after using 70% Alcohol and Tissue Cleaner, there was statistically significant in the significant level of 5% in both of them.

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Rhizosphere Inhibition of Cucumber Fusarium Wilt by Different Surfactinexcreting Strains of Bacillus subtilis

  • Jia, Ke;Gao, Yu-Han;Huang, Xiao-Qin;Guo, Rong-Jun;Li, Shi-Dong
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.140-151
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    • 2015
  • Bacillus subtilis B006 strain effectively suppresses the cucumber fusarium wilt caused by Fusarium oxysporum f. sp. cucumerinum (Foc). The population dynamics of Foc, strain B006 and its surfactin over-producing mutant B841 and surfactin-deficient mutant B1020, in the rhizosphere were determined under greenhouse conditions to elucidate the importance of the lipopeptides excreted by these strains in suppressing Foc. Results showed that B. subtilis strain B006 effectively suppressed the disease in natural soil by 42.9%, five weeks after transplanting, whereas B841 and B1020 suppressed the disease by only 22.6% and 7.1%, respectively. Quantitative PCR assays showed that effective colonization of strain B006 in the rhizosphere suppressed Foc propagation by more than 10 times both in nursery substrate and in field-infected soil. Reduction of Foc population at the cucumber stems in a range of $0.96log_{10}ng/g$ to $2.39log_{10}ng/g$ was attained at the third and the fifth weeks of B006 treatment in nursery substrate. In field-infected soil, all three treatments with B. subtilis suppressed Foc infection, indicated by the reduction of Foc population at a range of $2.91log_{10}ng/g$ to $3.36log_{10}ng/g$ at the stem base, one week after transplanting. This study reveals that the suppression of fusarium wilt disease is affected by the effective colonization of the surfactin-producing B. subtilis strain in the rhizosphere. These results improved our understanding of the biocontrol mechanism of the B. subtilis strain B006 in the natural soil and facilitate its application as biocontrol agent in the field.