• Journal of Microbiology and Biotechnology
• /
• 제8권1호
• /
• pp.14-20
• /
• 1998

Syntheses of the B. stearothermophilus xylanolytic enzymes such as xylanases, ${\beta}$-xylosidases, ${\alpha}$-arabinofurano-sidases, and esterases, were observed to be regulated by the carbon source present in the culture media. Xylan induced synthesis of ${\beta}$-xylosidase at the highest level while xylose gave about 30% of the ${\beta}$-xylosidase activity induced by xylan. The lowest syntheses of the xylanolytic enzymes above mentioned were detected in the basal medium containing glucose as a sole carbon source. When a mixture of xylan and glucose was used as a carbon source, we could observe glucose repression of xylanase (about 70-fold) and ${\beta}$-xylosidase (about 40-fold) syntheses. Whereas, the level of the glucose repression of the expression of the xylA gene encoding the major ${\beta}$-xylosidase of B. stearothermophilus was assessed to be about l0-fold when the relative amounts of the xylA transcript were determined. From the sequence of the xylA gene, we could find two CRE-like sequences (CRE-l: nucleotides ＋124 to ＋136 and CRE-2:＋247 to ＋259) within the reading frame of the xylA gene, either or both of which were suspected to be involved in catabolite repression of the xylA gene.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
• /
• pp.167-170
• /
• 2000

B. Stearothermophilus 단백질 분해 효소는 2가 금속 이온인 $Ca^{2+}$에 의하여 내열성이 향상되며 $75^{\circ}C$에서 최대 역가를 나타내었다. 2mM $Ca^{2+}$를 첨가하여 B. Stearothermophilus를 이용한 호열 ${\cdot}$ 호기성 소화공정 운전할 경우 각각 최대량 대비 51%와 27%의 DOC 및 세포외단백질의 분해가 이루어지는 것을 확인하였다.

• 한국식품위생안전성학회지
• /
• 제21권3호
• /
• pp.159-163
• /
• 2006

건포류 86건에 대한 시험 결과, 37건(43.0%=37/86)에서 Bacillus spp가 18건(20.9%=18/86)에서 B. cereus가 분리되었다. 분리 동정된 37주는 B. cereus 48.6%(18/37), B. mycoides 13.5%(5/37), B. coagulus 5.4%(2/37). B. firmus 5.4%(2/37), B. circulus 2.7%(1/37), B. stearothermophilus 2.7%(1/37), B. pumilus 2.7%(1/37), E. spp. 8.1%(3/37), 그리고 Brebacillus brevis 10.8%(4/37) 등으로 나타나 식중독균의 일종인 B. cereus가 가장 많이 나타나 주의를 요할 것으로 사료된다.

• 생명과학회지
• /
• 제14권5호
• /
• pp.778-787
• /
• 2004

유기물이 풍부한 토양에서 분리하여 동정한 Bacillus stearothermophilus DL-3와 미강을 사용하여 미생물 제재를 제조하고 미생물 제재의 처리가 상추(적치마 상추)와 배추(가락 신1호 배추)재배 토양의 미생물상에 미치는 영향 및 상추와 배추의 생장에 미치는 영향을 조사하였다. 미생물 제재를 처리 한 6주 후, 미생물 제재를 처 리 한 토양에 존재하는 토양 미생물의 총 균수는 미생물 제재를 처리하지 않은 토양에 존재하는 토양 미생물의 총 균수에 비하여 미생물 제재를 처리한 양에 비례하여 많음을 확인하였다. 미생물 제재를 처리한 토양에서 재배한 상추와 배추의 생육이 미생물 제재를 처리하지 않은 토양에서 재배한 상추와 배추에 비하여 빠름을 확인하였다. 미생물 제재를 처리한 토양에서는 처리하지 않은 토양에 비하여 전체적인 토양 미생물 수의 증가와 작물의 생장에 유용한 종류의 미생물이 상대적으로 증가하기 때문에 상추와 배추의 생장을 촉진시킨다고 판단된다.

• Journal of Microbiology and Biotechnology
• /
• 제12권4호
• /
• pp.683-686
• /
• 2002

To achieve optimum operation of a thermophilic aerobic digestion (TAD) process for waste activated sludge (WAS), TAD experiments using Bacillus stearothermophilus (ATCC 31197) were carried out to investigate the optimum concentration of dissolved oxygen (DO). TAD reactors were operated at DO concentrations of 0, 1, 2, 3, 4, and 5 ppm, and the results showed that the WAS could be successfully degraded by a TAD system operated with a DO concentration of 1 ppm and above. When the TAD system with an optimum additive (2 mM Ca ion), selected from a previous study, and 1 ppm DO concentration were combined with a thermal pretreatment ($121^{\circ}C$, 10 min), the results exhibited upgraded total suspended solids and an enhanced protein degradation.

• 한국수산과학회지
• /
• 제10권3호
• /
• pp.145-149
• /
• 1977

Thermal resistance of dried bacterial spores against dry heat was determined. Spare suspensions of Bacillus subtilis var. niger ATCC 9372, Bacillus stearothermophilus Oxoid Code BR 23 and Clostridium sporogenes ATCC 19404 were located on aluminium strips, dried in electric oven under vacuum at room temperature for 10 minutes. The aluminium strips were laid in the middle of gas flow (hot air and superheated steam) with the velocity of 6 m/sec and heated at $120^{\circ}C$ for 180 seconds. The calculated D-values showed that there were no remarkable differences in the heat resistance of bacterial spares between $R.H.\leqq0.012$ and R. H.=0.51. Furthermore the thermal resistance of B. subtilis spores to dry heat was greater than that of B. stearothermophilus.

• Journal of Microbiology and Biotechnology
• /
• 제10권6호
• /
• pp.764-769
• /
• 2000

The lipase gene(lip) from Bacillus stearothermophilus was recombined in vitro by utilizing the DNA shuffling technique. After four rounds of shuffling, transformation, and screening based on the initial rate of clear zone formation on a tricaprylin plate, a clone (M10) was isolated, the cell extract of which showed about 2.8-fold increased lipase activity. The DNA sequence of the mutant lipase gene (m10) showed 3 base changes, resulting in two cryptic mutations and one amino acid substitution: S113($AGC{\rightarrow}AGT$), L252 ($TTG{\rightarrow}TTA$), and G275E ($GGA{\rightarrow}GAA$). SDS-PAGE analysis revealed that the increased enzyme activity observed in M10 was partly caused by high expression of the m10 lipase gene. The amount of the expressed G275E lipase was estimated to comprise as much as 41% of the total soluble proteins of the cell. The maximum velocity ($V_{max}$) of the purified mutant enzyme for the hydrolysis of olive oil was measured to be 3,200 U/mg, which was 10% higher than that of the parental (WT) lipase (2,900 U/mg). Its optimum temperature for the hydrolysis of olive oil was $68^{\circ}C$ and it showed a typical $Ca^{2+}$-dependent thermostability, properties fo which were the same as those of the WT lipase. However, the mutant enzyme exhibited a high enantiospecificity towards (S)-naproxen compared with the WT lipase. In addition, it showed increased hydrolytic activity towards triolein, tricaprin, tricaprylin, and tricaproin.

• Journal of Microbiology and Biotechnology
• /
• 제13권3호
• /
• pp.451-456
• /
• 2003

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

• Applied Biological Chemistry
• /
• 제33권1호
• /
• pp.79-86
• /
• 1990

Bacillus stearothermophilus KFCC 21203를 배양하여 cyclodextrin(CD)을 분해하는 효소를 분리, 정제하고 정제효소의 몇 가지 특성을 조사하였다. 배양액에서 얻은 조효소를 염석, DEAE-cellulose column chromatography, Ultro AcA 34 gel filtration등의 방법으로 15배 정제하였으며 회수율은 77.2%이었다. 정제효소의 specific activity는 12.30units/mg protein 이었고 분자량은 약 29,500정도였다. 이 효소의 작용최적 PH는 5.5, 작용최적온도는 $55^{\circ}C$였으며 $40^{\circ}C$이하의 온도와 pH $5.0{\sim}8.0$의 범위에서 안정하였고 ${\gamma}-CD$에 대한 Km치는 $3.78{\times}10^{-3}$ M이었다. 정제효소는 ${\beta}-CD$에 대한 활성이 매우 낮았고 ${\alpha}-CD$에는 거의 활성이 없었으나 ${\gamma}-CD$에는 매우 높은 활성을 나타내었으며, 이의 분해산물로는 주로 glucose 및 maltose 그리고 소량의 maltotriose였다. 또한 amylose, potato starch, corn starch, amylopectin 및 maltooligomer등에는 높은 활성을, 그리고 glycogen, dextrin에도 비교적 높은 활성을 나타내었고 분해산물로는 주로 glucose와 maltose였다.

• 미생물학회지
• /
• 제30권4호
• /
• pp.305-309
• /
• 1992

Bacillus stearothermophilus 의 cytosine deaminase (EC 3. 5. 4.1 )를 수율 52.7% 로 7.2배 부분정제했다. 부분정제된 cytosine deaminase 는 cytosine 을 유일한 기질로 이용하였으머 5-methylcytosine 과 5-fluorocytosine 은 기질고 이용되지 못햇으며 cytosine 에 대한 Michaelis 정수 Km 값은 5.9 mM 이었다. 본효소는 pH 4.0 에서 7.0 까지의 폭 넓은 pH 영역에서 안정했으며 80.deg.C 에서 10 분간 열처리하여도 75% 이상의 효소활성이 잔존하여 높은 내열성 효소였다. 본 효소의 반응최적 pH 는 7.0-7.5 였으며 반응 최적 온도는 35. 37.deg.C 였다. 그리고 Arrhenium plot 에 의하여 계산된 활성화 에너지 값(Ea value) 은 26 Kcal/mol 이었다. 본 효소는 중금속인 1 mM의 $Cd^{2+}$ , $Hg^{2+}$ 및 $Cu^{2+}$ 에 의하여 완정히 실활되었으며 GMP 와 CMP 에 의해서는 효소활성이 촉진되었다. 특히 본 효소는 p-chloromercuribenzoate 에 의하여 효소 활성이 강하게 저해되어 thiol 효소임을 추정할 수 있었다.