• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.147-152
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• 2009

A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.

• Journal of Microbiology and Biotechnology
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• v.5 no.1
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• pp.18-21
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• 1995

Endoglucanase gene of Pseudomonas sp. LBC505 was previously cloned in pUCl9 to yield plasmid pLC1. overproduction of endoglucanase was attempted by following ways. First, the endoglucanase gene of Pseudomonas sp. LBC505 cloned in pUCl9(pLC1) was tandemly inserted, step by step, into a expression vector pKK223-3 in a directly repeated form to enhance productivity of endoglucanase. Escherichia coli containing pKCC30 among the resulting plasm ids showed the higher yield of the endoglucanase. Ecoli harboring pKCC30 which had three inserted endoglucanase genes expressed about 12.3 times as much CMCase activity as Ecoli harboring pLCl. Second, the endoglucanase gene was subcloned into Bacillus subtilis expression vector pgnt41 for both overproduction and extracellular secretion of the endoglucanase. A resulting plasmid pgntc15 in Bacillus subtilis expressed 4.3-fold higher levels of CMCase activity than that of E.coli harboring pLCl and the endoglucanase produced was entirely secreted into the culture medium.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.573-579
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• 1995

A Bacillus strain capable of producing an extracellular malto-oligosaccharides forming $\alpha $-amylase was isolated from soil and designated as Bacillus sp. SUH4-2. The enzyme was purified by ammonium sulfate fractionation, DEAE-Toyopearl and Mono-Q HR 5/5 column chromatographies using a FPLC system. The specific activity of the enzyme was increased by 16.1-fold and the yield was 13.5%. The optimum temperature for the activity of $\alpha $-amylase was 60-65$\circ$C and more than 50% of initial activity was retained after the enzyme was incubated at 60$\circ$C for 40 min. The enzyme was stable over a broad pH range of 5.0-8.0 and the optimum pH was 5.0-6.0. The molecular weight of the enzyme was determined to be about 63.6 kD and isoelectric point was around 5.8. The enzyme activity was strongly inhibited by Mn$^{2+}$, Ni$^{2+}$, and Cu$^{2+}$ ; slightly by Ca$^{2+}$. The purified enzyme produced starch hydrolyzates containing mainly maltose and maltotriose from soluble starch. The starch hydrolyzates were composed of 11% glucose, 59% maltose, 25% maltotriose and 5% maltotetraose.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.186-191
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• 2005

In this study, a microorganism-produced protease was used to improve the quality of fabrics. First, the protease-producing bacteria were isolated from soils, and one of them was selected and identified as Bacillus sp. SJ-121. The optimal medium composition for its growth and protease production was determined to be as follows: glucose 1g/L, soybean meal 0.5g/L, soy peptone 0.5, $K_2HPO_4\;0.2,\;MgSO_4\cdot7H_2O\; 0.002,\;NaCl\;0.002,\;and\;Na_2CO_3g/L$. Also, the optimal temperature for the production of the protease by Bacillus sp. SJ-121 was about $40^{\circ}C$ at pH 7. The wool and silk were treated with the protease from Bacillus sp. SJ-121. Following the protease treatment, changes in the surface of a single yarn of the fabrics were observed by both an optical microscope and a scanning electron microscope (SEM). Changes in the K/S value of the wool and silk were measured by spectrophotometric analysis, in order to determine the amount of dye uptake in the fabrics. We also performed a tensile strength examination in order to determine the degree and nature of mechanical changes in single yarns of the wool and silk fabrics. By increasing the protease treatment time to 48 h, the dyeing characteristics of the fabrics were enhanced, and the surfaces of the single yarns of the fabrics became smoother, due to the removal of soil and scale in them. However, no mechanical changes were detected in the fabrics. Therefore, we suggest that proper treatment of the protease produced by Bacillus sp. can improve the quality of silk and wool.

• Applied Biological Chemistry
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• v.43 no.1
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• pp.1-6
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• 2000

The quality changes of chunggugjang produced by Bacillus sp. CS-17 was investigated with fermentation time. The pH was gradually alkalized. L-value became low while b-value became high. Both strength and hardness extraordinarily decreased during fermentation. Total content of free amino acid was $8274.7{\sim}9301.4\;mg%$ and phenylalanine, lysine, leucine and tyrosine were the most abundant components among the amino acids. The ratio of essential amino acid was $66.6{\sim}71.8%$. As a result of sensory test, it was found that the chunggugjang fermented by Bacillus sp. CS-17 was suitable enough to be produced for commercial purpose.

• Food Science and Preservation
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• v.8 no.1
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• pp.92-101
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• 2001

In order to develop the Production and application of cholesterol oxidase, a cholesterol degradation bacteria which produces a remarkable amount of extracellular cholesterol oxidase has been isolated from Korea traditional salt fermented flat fish. The isolated strain was identified as a strain of Bacil1us sp. based on its morphological, physiological characteristics and cellular fatty acid compositions. Experiments were carried out to optimize the condition of cholesterol oxidase production using Bacillus sp. SFF34. Bacillus sp. SFF34 was shown to give the maximum yield of cholesterol oxidase in the medium containing 2.0% glucose, 0.5% yeast extract, 0.02% MgSO$_4$$.$7H$_2$O, 0.025% K$_2$HPO$_4$, 0.15% NH$_4$NO$_3$ and 0.2% cholesterol. The optimum culture conditions, temperature, initial pH and agitation speed were 30$^{\circ}C$, 7.0 and 150rpm respectively. The enzyme production reached a maximum level at 24 hrs of cultivation(2.42 U/ml).

• Journal of the Society of Cosmetic Scientists of Korea
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• v.26 no.1
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• pp.107-124
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• 2000

A bacterial strain No. HB-5, which was capable of producing a pretense in the culture conditions, was isolated from the soil . The pretense was purified from cultural filtrate of Bacillus sp. HB-5 by membrane ultrafiltration and DEAE- cellulose chromatography, gel filtration on Sephadex G-100. The molecular weight was estimated to be 60k4a. The optimal pH and temperature for the activity of the purified pretense pH were 11 and 5$0^{\circ}C$ , respectively. The enzyme was stable within a pH range 8-12 and up to 6$0^{\circ}C$ . The enzyme activity was highly inhibited by PMSF at 1mM. The proteolytic actions of pretense and papain on human epidermis keratins which are major protein impurities on the skin, were compared. The bacterial pretense degraded more effectively than papain. Product containing 2% protease exhibited 21% increase on the skin coloration index. These results suggest that cosmetic product containing pretense produced by Bacillus sp. HB-5 could remove the adherent keratin layer and then make a softer skin.

• Journal of Korean Society of Environmental Engineers
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• v.29 no.2
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• pp.169-175
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• 2007

The Bio Best Bacillus(B3) and Rotating Activated Bacillus Contactor(RABC) processes, in which Bacillus strains are predominating, are reported to remove nitrogen and phosphorus as well as organic matter effectively. Nevertheless the nutrient removal characteristics of the Bacillus strains have not been studied in detail so far. This study investigated the organic and nutrient removal by Bacillus strains, Bacillus megaterium(KCTC 3007), Paenibacillus polymyxa(KCTC 3627), and Bacillus sp. A12, C21, F12, and L1(isolated from a B3 process), by incubating the strains in 0.2% nutrient broth at $30^{\circ}C$. Burkholderia cepacia(KCTC 2966), a common activated sludge organism, was used as a reference species for comparison. Although the degradation rate was affected by the population sire, the specific removal rates of organic matter by Bacillus strains were greater by $2\sim5$ times than that of Burkholderia. In particular, the culture bottles inoculated with the endospores of Bacillus megaterium and Bacillus sp. C21, F12, and N12 showed significantly higher degradation rate than those of vegetative cells.

• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.148-153
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• 1989

A strain of alkalophilic Bacillus sp. YC-335 has been isolated from soil. The strain was capable of producing large amount of cyclodextrin glycosyl transferase (CGTase) in the culture broth. The preferable medium composition has been determined to be as follows : 1.5% soluble starch, 5% corn steep liquor, 0.1% $K_2$HPO$_4$, 0.02%mgSO$_4$.7$H_2O$, 1% CaCO$_3$and 1% Na$_2$CO$_3$(pH 10.3). The highest enzyme production was observed after 48 hours of cultivation at 31$^{\circ}C$. The optimum pH and temperature for the activity of crude enzyme were 6.0 and 5$0^{\circ}C$, respectively. The enzyme was stable between pH 5 and 9, and upto 5$0^{\circ}C$. The enzyme converted starch into $\alpha$-, $\beta$- and ${\gamma}$-CD in the relative amounts of 1:10:1.5, respectively.