Isolation and identification of two novel SDS-resistant secreted chitinases from Aeromonas schubertii.
Two SDS-resistant endochitinases, designated as ASCHI53 and ASCHI61, were isolated from Aeromonas schubertii in a soil sample from southern Taiwan. MALDI-TOF mass measurement indicates the molecular weights of 53,527 for ASCHI53 and 61,202 for ASCHI61. N-terminal and internal amino acid sequences were obtained, and BLAST analysis of the sequences and MS/MS peptide sequencing showed that they were novel proteins. Degradation of chitin by these two endochitinases gave rise to hexameric chitin oligosaccharide, a compound known to have several potent biomedical functions. ASCHI53 and ASCHI61 retained, respectively, 65% and 75%, of their chitinase activity in the presence of 5% SDS and 100% of their activity in the presence of 10% beta-mercaptoethanol. These results demonstrate that they are SDS-resistant endochitinases and probably have a rigid structure.
Liu
Chao-Lin
CL
Graduate School of Biochemical Engineering, and Dept. of Safety Health and Environmental Engineering, MingChi University of Technology, Taipei, Taiwan, Republic of China. f2402002@ms16.hinet.net
Shen
Chia-Rui
CR
Hsu
Fong-Fu
FF
Chen
Jeen-Kuan
JK
Wu
Pei-Tzu
PT
Guo
Shang-Hsin
SH
Lee
Wen-Chien
WC
Yu
Feng-Wei
FW
Mackey
Zachary B
ZB
Turk
John
J
Gross
Michael L
ML
D000333:Aeromonas / Q000201:enzymology*; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D002688:Chitinases / Q000302:isolation & purification* / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D001426:Bacterial Proteins; D012967:Sodium Dodecyl Sulfate; D002688:Chitinases