CLONING AND EXPRESSION OF CLOSTRIDIUM-THERMOCELLUM GENES-CODING FOR THERMOSTABLE EXOGLUCANASES (CELLOBIOHYDROLASES) IN ESCHERICHIA-COLI-CELLS

被引：32

作者：

TUKA, K ^{[1
]}

ZVERLOV, VV ^{[1
]}

BUMAZKIN, BK ^{[1
]}

VELIKODVORSKAYA, GA ^{[1
]}

STRONGIN, AY ^{[1
]}

机构：

[1] ACAD SCI USSR,INST MOLEC GENET,KURCHATOV SQ 46,MOSCOW 123182,USSR

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1990年 / 169卷 / 03期

关键词：

D O I：

10.1016/0006-291X(90)92001-G

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

By special screening approach two independent Cl. thermocellum genes directing the synthesis of thermostable glucanases with an exo-mode of action have been isolated from pUC19-based gene bank in E. coli TG1. The genes are located on 3, 4 and 11,3 kb DNA fragments showing no homology. E. coli-derived exoglucanases, presumably, cellobiohydrolases, are able to cleave lichenan, carboxymethyl cellulose, xylan and p-nitrophenyl derivatives of cellobioside and lactoside. Cellobiose is the main degradation product of carboxymethyl cellulose, treated with the identified exoglucanases. With p-nitrophenil-β-D-cellobioside as substrate the enzymes had a pH optimum around 6,5 and a temperature optimum at 65°C. The identified and expressed enzymes differ from all other Cl. thermocellum proteins known to date. © 1990.

引用

页码：1055 / 1060

页数：6

共 20 条

[1]

AIT N, 1982, J GEN MICROBIOL, V128, P569

[2]

BEGUIN P, 1985, J BACTERIOL, V162, P102

[3]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[4]

BUMAZKIN BK, 1990, IN PRESS BBRC

[5] CHARACTERIZATION OF 2 CEL (CELLULOSE DEGRADATION) GENES OF CLOSTRIDIUM-THERMOCELLUM CODING FOR ENDOGLUCANASES [J].

CORNET, P ;

MILLET, J ;

BEGUIN, P ;

AUBERT, JP .

BIO-TECHNOLOGY, 1983, 1 (07) :589-594

[6] SEQUENCE OF THE CELLULASE GENE OF CLOSTRIDIUM-THERMOCELLUM CODING FOR ENDOGLUCANASE-B [J].

GREPINET, O ;

BEGUIN, P .

NUCLEIC ACIDS RESEARCH, 1986, 14 (04) :1791-1799

[7]

HALLIWELL N, 1989, BIOTECHNOL APPL BIOC, V11, P201

[8] SUBSTRATE SPECIFICITIES OF EXO-TYPE AND ENDO-TYPE CELLULASES IN THE HYDROLYSIS OF BETA-(1-]3)-MIXED AND BETA-(1-]4)-MIXED D-GLUCANS [J].

KANDA, T ;

YATOMI, H ;

MAKISHIMA, S ;

AMANO, Y ;

NISIZAWA, K .

JOURNAL OF BIOCHEMISTRY, 1989, 105 (01) :127-132

[9]

Maniatis T., 1982, MOL CLONING

[10]

MILLER GL, 1960, ANAL BIOCHEM, V2, P127, DOI DOI 10.1016/0003-2697(60)90004-X

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