Cloning, sequences, and characterization of two chitinase genes from the antarctic Arthrobacter sp strain TAD20:: Isolation and partial characterization of the enzymes

被引:54
作者
Lonhienne, T
Mavromatis, K
Vorgias, CE
Buchon, L
Gerday, C
Bouriotis, V
机构
[1] Univ Crete, Dept Biol, Div Appl Biol & Biotechnol, Iraklion 71110, Crete, Greece
[2] Univ Liege, Inst Chem B6, Biochem Lab, B-4000 Liege, Belgium
[3] Univ Crete, Inst Mol Biol & Biotechnol, Enzyme Technol Div, Iraklion 71110, Crete, Greece
[4] Univ Athens, Fac Biol, Dept Biochem Mol Biol, Athens 15701, Greece
[5] Inst Univ Technol, Lab Microbiol Froid, F-27000 Evreux, France
关键词
D O I
10.1128/JB.183.5.1773-1779.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Arthrobacter sp. strain TAD20, a chitinolytic gram-positive organism, was isolated from the sea bottom along the Antarctic ice shell. Arthrobacter sp. strain TAD20 secretes two major chitinases, ChiA and ChiB (ArChiA and ArChiB), in response to chitin induction. A single chromosomal DNA fragment containing the genes coding for both chitinases was cloned in Escherichia coli. DNA sequencing analysis of this fragment revealed two contiguous open reading frames coding for the precursors of ArChiA (881 amino acids [aa]) and ArChiB (578 aa). ArChiA and ArChiB are modular enzymes consisting of a glycosyl-hydrolase family 18 catalytic domain as well as two and one chitin-binding domains, respectively. The catalytic domain of ArChiA exhibits 55% identity with a chitodextrinase from Vibro furnissii. The ArChiB catalytic domain exhibits 33% identity with chitinase A of Bacillus circulans. The ArChiA chitin-binding domains are homologous to the chitin-binding domain of ArChiB. ArChiA and ArChiB were purified to homogeneity from the native Arthrobacter strain and partially characterized. Thermal unfolding of ArChiA, ArChiB, and chitinase A of Serratia marcescens was studied using differential scanning calorimetry. ArChiA and ArChiB, compared to their mesophilic counterpart, exhibited increased heat lability, similar to other cold-adapted enzymes.
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页码:1773 / 1779
页数:7
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