• 한국미생물·생명공학회지
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• 제40권4호
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• pp.303-309
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• 2012

Keratinases are of particular interest because of their action on insoluble keratins and generally on a broad range of protein substrates. Alkalophilic and neutrophilic actinomycete strains isolated from different soil samples, rich in keratinaceous substances were screened for keratinolytic activity. An alkalophilic isolate, TBG-S13A5, was found to possess good keratinolytic activity and was able to utilize feather as the sole carbon and nitrogen source. TBG-S13A5 exhibited an off-white aerial mass color with a rectus-flexibilis type of spore chain. The morphological, microscopical and biochemical characters were comparable with that of Streptomyces albidoflavus. Fatty acid methyl ester profiling (FAME) and 16S rDNA sequence analysis confirmed its identity as a strain of S. albidoflavus. Under submerged fermentation conditions, maximum protease production was recorded on the $5^{th}$ day of incubation at $30^{\circ}C$, using basal broth of pH 9.0 with 0.25% (w/v) white chicken feather. This strain could affect feather degradation when the initial pH was 8 and above and maximum protease production was recorded when the initial pH was around 10.5. The effectiveness of the crude enzyme in destaining and leather dehairing were also demonstrated.

• 한국미생물·생명공학회지
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• 제17권3호
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• pp.213-218
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• 1989

Cloning을 위한 host와 vector의 이용 가능성을 타진하기 위해 호알칼리성 Bacillus속 K-17을 host로, pUB110과 pBD64를 vector로 사용하여 Bacillus속 K-17의 protoplast 형질전환조건을 검토하였다. 원형질체의 형성은 200$\mu\textrm{g}$/$m\ell$의 Iysozyme 을 처리하였으며, 원형질체 형성의 최적 온도, PH및 배양시간은 각각 4$0^{\circ}C$, 7.0 및 4시간으로 나타났다. 원형질체는 DM3 재생배지에서 재생시켰으며 0.8% agar, 0.5M sodium succinate 농도에서 가장재생이 좋았다. 형질전환시 PEG의 농도는 40%(w/v) PEG 6,000 30%(v/v)가 최적으로 나타났다. 형질전환체의 특성을 조사한 결과, plasmid 안정성은 pUB110이 pBD64보다 더 안정하였으며, 최대 효소활성은 비슷하였지만 효소 분비시간은 pUB110 은 2.5일, pBD64의 경우는 3일로 Bacillus속 K-17의 2일에 비해 약간 지연되었다.

• 한국염색가공학회지
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• 제22권2호
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• pp.145-154
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• 2010

An alkalophilic microorganism capable of degrading dyes was developed for the treatment of alkaline dye solution. This strain was identified as Pseudomonas species. Using this microorganism, biological treatment of dye was studied in Erlenmeyer flasks. The characteristics of this microorganism were observed under various incubating-condition such as temperature, pH, nitrogen source, and macronutrients concentration. The removal effciencies of Disperse Red 60 from synthetic wastewater were 33.5 ~ 36.9% at the range of $30{\sim}40^{\circ}C$, and they were 31.1 ~ 36.7% at the range of initial pH 8 ~ pH 10, respectively. The optimal culture medium was found to be 0.25%(w/v) yeast extract, 0.25%(w/v) polypeptone, 0.1%(w/v) $KH_2PO_4$, 0.2%(w/v) $MgSO_4{\cdot}7H_2O$, and 1.0%(w/v) $Na_2CO_3$. In treatment of various dyes using Erlenmeyer flasks, the removal effciencies of Disperse Blue 87, Disperse Yellow 64, Disperse Red 60, Acid Blue 193, Acid Red 138, and Direct Yellow 23 were found to be 76%, 71%, 58%, 93%, 94%, and 90% respectively after 24hrs reaction of alkalophilic strain Pseudomonas sp. YBE-12.

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.524-528
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• 1989

토양으로부터 분리한 호알카리성 Bucillus sp. YS-309는 $\beta$-galactosidase 생산력이 강력하였으며 중성배지 보다도 알카리배지 중에서 균체생육 및 효소생산량이 많았다. 이 분리균의 최적 배양조건은 lactose 0.5%, yeast extract 0.5%, defatted soybean meal extract 0.6%(v/v)가 함유된 PH9.9의 알카리 배지를 사용하였을 때 얻어졌다. 이 때의 배양온도는 4$0^{\circ}C$, 배양시간은 24-30시간의 바람직한 것으로 나타났다. 또 IPTG와 lactose에 의해 효소생산이 유도적으로 증가하였으며 전자는 후자의 3배 효과가 있었다. Lactose 농도도 균체생육 및 효소생산에 영향을 미쳤으며 그 농도가 1.5% 이상 배지 중에 첨가되면 균체생육 및 효소생산이 모두 감소하였다.

• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.17-21
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• 1991

A gene coding for ${\beta}-xylosidase$ in alkalophilic Bacillus sp. YC-335 isolated from soil was cloned into Escherichia coli HB101 using plasmid pBR322. The recombinant plasmid pYK40 was isolated, and the cloned HindIII fragment was 15 kilobases (kb). To reduce the size of the inserted DNA fragment of pYK40, the 15 kb HindIII fragment was subjected to a series of subclonings. A 6 kb subfragment was found to code for ${\beta}-xylosidase$ activity, and the recombinant plasmid was named pYK44. Southern hybridization analysis revealed that the cloned gene hybridized with 3.5 kb, 1.5 kb, and 1.0 kb of HindIII cleaved chromosomal DNA from Bacillus sp. YC-335. ${\beta}-xylosidase$ activity produced by recombinant E. coli was found to be 11 times higher than that produced by Bacillus sp. YC-335. Xylan was required to induce the production of ${\beta}-xylosidase$ in Bacillus sp. YC-335.

• 한국미생물·생명공학회지
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• 제19권6호
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• pp.543-547
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• 1991

토양으로부터 호알칼리성 미생물을 분리하여 동정 한 결과, Bacillus속의 특성을 나타내어 Bacillus sp. SH-8로 명명하였다. Bacillus sp. SH-8은 초기 pH 9.0이상의 알칼리에서만 생육이 가능하였으며 중성 pH에서는 불가능하였다. Bacillus sp. SH-8로부터 U.V.조사에 의해 중성배지에서도 생육이 가능한 변이주를 분리하여 Bacillus sp. SH-8M으로 명명하였으며, 초기 pH 6.9부터 11.0까지 생육이 가능하였다. 따라서 이들 두 균주는 pH 변화에 따른 생육과 효소생산을 비교, 검토하는데 적합한 균주로 생각된다.

• 한국미생물·생명공학회지
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• 제17권6호
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• pp.563-567
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• 1989

알칼리성 세균으로부터 ammonium fumarate를 이용하여 글루탐산을 생성할 수 있는 균주를 분리, 동정하였으며, 글루탐산 생성에 미치는 여러가지 요인들과 생성기전에 관하여 조사하였다. 선택된 균주는 형태, 성질 등에서 Bacillus brevis와 유사하였다. 여러가지 배양조건 중에서 2% fumaric acid에 0.8% nutrient broth 를 첨가하고 암모니아수로 pH를 9로 조절한 배지에서 글루탐산 생성이 가장 좋았으며, 1% fumaric acid로부터 글루탐산으로의 전환율은 약 22%였다. 생성기전을 알아보기 위한 실험에서 whole cell을 이용하였을 때 ammonium fumarate로부터 글루탐산 생성을 볼 수 있었으며, $\alpha$-ketoglutaric acid와 aspartic acid 가 포함된 반응액에서 glutamic acid 생성과 aspartic acid 소실을 동시에 볼 수 있었다. 따라서 생성기전은 fumaric acid가 aspartic acid로 전환되고, aspartic acid와 $\alpha$-KGA가 반응하여 glutamic acid가 생성되는 것으로 추정된다.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.863-869
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• 2001

A systematic study of microbial fuel cells comprised of alkalophilic Bacillus sp. B-31 has been carried out under various operating conditions. A significant amount of electricity was generated when redox mediators were used. Among the phenothiazine-type redox dyes tested, azure A was found to be the most effective both in maintaining a high cell voltage and for the long-term operation. The maximum efficiency was and for the long-term operation. The maximum efficiency was obtained at ca. $50^{\circ}C$ giving an open circuit voltage of 0.7V. A small change in temperature did not significantly affect the cell performance, but a rapid decrease in performance was observed below $20^{\circ}C$ and above $70^{\circ}C$. It was noticeable that fuel cell efficiency and discharge pattern depended strongly on the carbon source used in the initial culture medium. Regardless of the initial carbon sources, only glucose and trehalose were utilized as substrates. Galactose, however, was not substantially utilized except when galactose was used in the initial medium. Glucose, in particular, showed $87\%$ coulombic efficiency, which was the highest value ever reported, when Bacillus sp. was cultured in a maltose-containing medium. This study demonstrates that highly efficient microbial fuel cells can be constructed with alkalophilic microorganisms by fine-tuning the operating conditions and by carefully selecting carbon sources in the initial culture medium.

• 한국미생물·생명공학회지
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• 제18권1호
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• pp.44-48
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• 1990

토양에서 분리한 호알카리성 Bacillus sp. YC-335가 생산하는 CGTase를 ethanol 침전법, DEAE-Toyopearl column chromatography, Sephadex G-100 column chromatography 방법 등에 의해 대량 정제 하였다. 이때 수율은 17.8 이었고 52.9배의 정제도를 가진 효소단백질을 얻을 수 있었다. 정제효소는 SDS-polyacrylamide gel 전지영동에 의해 분자량이 75,000 정도인 단일 peptide의 단백질임을 확인하였고 효소의 최적활성 pH는 6.0 있었으며 pH 안정성은 pH 6-10까지 안정하였다. 최적활성온도는 5$0^{\circ}C$이었으며 열안정성은 5$0^{\circ}C$까지 안정하였고 이는 15mM CaCl$_2$에 의해 열안정성이 보호되었다.

• 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.