• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.14-20
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• 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
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• 2000.11a
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• pp.167-170
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• 2000

Proteolytic hydrolysis is one of the main enzymatic reaction of waste activated sludge (WAS) digestion. Pretense excreted from Bacillus stearothermophilus (ATCC 31197) showed optimum temperature of $75^{\circ}C$ for maxium heat stable proteolytic activity against azo casein. The dependency of $Ca^{2+}$, $Zn^{2+}$ on heat stability of proteolytic enzymes were measured with various concentrations. It was shown that $Ca^{2+}$ ion enhanced heat stability of these enzymes. Then thermophilic aerobic digestion (TAD) was performed using B. sterothermophilus with the addition of divalent ions. Performance of TAD process with ATCC 31197 activated by $Ca^{2+}$, $Zn^{2+}$ions in terms of dissolved organic carbon (DOC) concentration, extracellular protein concentration, and scanning electrion microscopy (SEM) analysis. The best result of protein reduction concentration in digestion test was obtained with the addition of 2 mM $Ca^{2+}$ ion.

• Journal of Food Hygiene and Safety
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• v.21 no.3
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• pp.159-163
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• 2006

37 Bacillus spp strains(43.0%=37/86), and 18 B. cereus(20.9%=18/86) were isolated on 86 dried marine products. Each isolates were 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) B. spp. 8.1%(3/37), and Brebacillus brevis 10.8%(4/37) etc.

• Journal of Life Science
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• v.14 no.5
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• pp.778-787
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• 2004

Effect of the microbial product made of Bacillus stearothermophilus DL-3, which was isolated from the soil and identified in this study, and rice bran on microorganisms in soil and growth of lettuce (Red skirt lettuce) and Chinese cabbage (Ga rack new No.1 Chinese cabbage) was investigated. Total numbers of microorganisms in the pot with untreated soil, treated with standard amount of microbial product and treated with double amounts of microbial product for growth of lettuce after 6 weeks were 2.78${\times}$10$^{7}$ CFU/g, 2.72${\times}$10$^{8}$ CFU/g and 3.63${\times}$10$^{8}$ CFU/g. Total numbers of microorganisms in the soil without treatment of microbial product and treated with standard amount of microbial product were 2.06${\times}$10$^{8}$ CFU/g and 5.49${\times}$10$^{8}$ CFU/g. Total numbers of microorganisms in the pot with untreated soil, treated with standard amount of microbial product and treated with double amounts of microbial product for growth of Chinse cabbage after 6 weeks were 1.43${\times}$10$^{7}$ CFU/g, 3.42${\times}$10$^{8}$ CFU/g and 7.22${\times}$10$^{8}$ CFU/g. Total numbers of microorganisms in the soil without treatment of microbial product and treated with standard amount of microbial product were 5.75${\times}$10$^{8}$ CFU/g and 7.96${\times}$10$^{8}$ CFU/g. On basis of leaf length, leaf width, leaf number, wet weight and dry weight, the growth of lettuce and Chinese cabbage on the soil treated with microbial product was faster than that on the untreated soil. The treatment of microbial product in the soil resulted in the increase of useful microorganisms, which seemed to enhance the growth of lettuce and Chinese cabbage.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.683-686
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• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.3
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• pp.145-149
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• 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
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• v.10 no.6
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• pp.764-769
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• 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
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• v.13 no.3
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• pp.451-456
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• 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
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• v.33 no.1
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• pp.79-86
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• 1990

Cyclodextrin hydrolase from Bacillus stearothermophilus KFCC 21203 was purified and the properties of the purified enzyme were investigated. The enzyme was purified 15 folds with 77 % recovery by ammonium sulfate fractionation, DEAE-cellulose chromatography, and Ultro AcA 34 gel filtration. The specific activity and the molecular weight of the enzyme were 1.30 units/mg protein and about 29,500, respectively, The maximum activity of the enzyme was shown at $55^{\circ}C$ and pH 5.5. However, stable temperature and pH were $40^{\circ}C$ and $5.0{\sim}8.0$, respectively. The Km value for ${\gamma}-cyclodextrin$ was $3.78{\times}10^{-3}$ M. The degradation activity of the enzyme was selectively high for ${\gamma}-cyclodextrin$, and very low for ${\beta}-cyclodextrin$, but not for ${\alpha}-cyclodextrin$. The decomposed products of ${\gamma}-cyclodextrin$ were mainly glucose and maltose, and a little mlatotriose. The activity of the enzyme was very high for amylose, potato starch, corn starch, amylopectin and maltooligomer, and relatively high for glycogen and dextrin. The decomposed products of them were mainly glucose and maltose.

• Korean Journal of Microbiology
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• v.30 no.4
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• pp.305-309
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• 1992

Cytosine deaminase (EC 3.5.4.1) from BaciNus stc~urorhermophilus was partially purified 7.2-fold with an overall yield of 52.7%. The partially purified enzyme deiiminated cytosine only.but not 5-methylcytosine and 5-fluorocytosine. The apparent Michaclis constant. Km valuefor cytosine was 5.9 mM. The enzyme was relatively stable in the range of pH 4.0 to 7.0.furthermore extremely thermo-stable : more than 75'X) of the activity was remained afterheating at 80$^{\circ}$C for I0 min at pH 6.5. The enzyme had a pH optimum at around pH7.0 to 7.5. and temperature optimum at 35 to 31$^{\circ}$C. And the activation energ (En value)determined from an Arrhenius plot was 26 Kcal/mol. The enzyme activity was stronglyinhibited by heavy metal ions such as Cd", Hg". Cut' at 1 mM, anJ by o-phenanthroline,and p-chloromcrcuribcnzoate at I mM. But the enrymc activity was activatetl increased byGMP, and CMP at 1 mM.ased by GMP, and CMP at 1 mM.