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키토산분해효소를 생산 분비하는 Bacillus sp . P16의 선발 및 특성
박노동,정미라,조유영,지연태 ( Ro Dong Park,Mi Ra Jung,Yoo Young Jo,Yeon Tae Chi ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.5
An endochitosanase-producing bacterium was isolated from soil and identified as a strain of Bacillus sp. The isolate was gram positive, rod shape (0.4-0.6×1.6-2.2 ㎛), endospore-forming, catalase positive, and mobility positive, and grown at pH 4.5-11.0 and upto 42℃ in the medium containing 2% NaCl. RAPD analysis of the DNA purified from the strain was also performed, and the chitosanase-producing strain was named as Bacillus sp. P16. The culture supernatant of the strain showed strong liquefaction activity and rapidly decreased viscosity of chitosan solution. By TLC and HPLC, chitooligosaccharides of DP 2-7 were separated and identified from the enzyme hydrolyzates of chitosan. The chitosanase from Bacillus sp. P16 was thus regarded as an endo-splitting type.
키토산분해효소 생산을 위한 Bacillus sp. P16 배양조건의 최적화
정미라,박노동,지연태,조유영 한국농화학회 1999 Applied Biological Chemistry (Appl Biol Chem) Vol.42 No.3
The optimal culture condition of Bacillus sp. P16 was investigated for production of an extracellular endo-splitting chitosanase. The best carbon and nitrogen sources for the chitosanase production were chitosan and tryptone, respectively. The best condition for the maximum activity was at 37℃ in a medium containing 0.5% powdered chitosan, 1% tryptone, and 1% NaCl(at initial pH 7.0) in a rotary shaker(200 rpm). In a jar fermenter, the culture duration shortened to 6∼12 hr for maximum activity and the enzyme activity increased about 100% compared with that of flask culture.
정미라,조유영,지연태,박노동 한국키틴키토산학회 1998 한국키틴키토산학회지 Vol.3 No.1
In order to obtain microbial endochitosanases for enzymatic production of high dp chitooligosaccharides from chitosan we screended 9 microbes which showed high chitinolytic activities. We tested the chitosanase activities of total 32 enzymes including 23 commercial enzymes and 9 crude enzymes produced by the microbes, in terms of the decreases in turbidity and viscosity of chitosan solution, the amount of pecipitation after neutralization, and reducing sugar-producing activity, after incubation with each enzyme. We selected Bacillus spp. P20, P21, and P30 as potential sources for production of endochitosanases to produce high dp chitooligosaccharides. Among them, enzyme produced by P2l was most potent and comparable to commercial enzyme E7. The culture supernatant of the strain P2l showed strong liquefaction activity and rapid decrease in viscosity of chitosan solution. By TLC and HPLC, chitooligosaccharides of dp 2-7 were separated and identified from the enzyme hydrolyzates of chitosn. The chitosanase from Bacillus sp. P2l was thus regarded as an endo-splitting and applicable to enzymatic production of high dp chitooligasaccharides from chitosan.