Isolation, purification, and enzymatic characterization of extracellular chitosanase from marine bacterium Bacillus subtilis CH2.
A Bacillus subtilis strain was isolated from the intestine of Sebastiscus marmoratus (scorpion fish) that was identified as Bacillus subtilis CH2 by morphological, biochemical, and genetic analyses. The chitosanase of Bacillus subtilis CH2 was best induced by fructose and not induced with chitosan, unlike other chitosanases. The strain was incubated in LB broth, and the chitosanase secreted into the medium was concentrated with ammonium sulfate precipitation and purified by gel permeation chromatography. The molecular mass of the purified chitosanase was detected as 29 kDa. The optimum pH and temperature of the purified chitosanase were 5.5 and 60°C, respectively. The purified chitosanase was continuously thermostable at 40°C. The specific acitivity of the purified chitosanase was 161 units/mg. The N-terminal amino acid sequence was analyzed for future study.
Korea Ocean Research and Development Institute, Ansan 426-744, Republic of Korea.
Chulhong
C
Oh
Mahanama
M
De Zoysa
Do-Hyung
DH
Kang
Youngdeuk
Y
Lee
Ilson
I
Whang
Chamilani
C
Nikapitiya
Soo-Jin
SJ
Heo
Kon-Tak
KT
Yoon
Abu
A
Affan
Jehee
J
Lee
D000818:Animals; D001412:Bacillus subtilis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D005110:Extracellular Space / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D005399:Fishes; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D007422:Intestines / Q000382:microbiology; D008969:Molecular Sequence Data; D012623:Seawater / Q000382:microbiology*; D013379:Substrate Specificity
D001426:Bacterial Proteins; D006026:Glycoside Hydrolases; C019868:chitosanase