CLONING, SEQUENCING AND ANALYSIS OF EXPRESSION OF A BUTYRIVIBRIO-FIBRISOLVENS GENE ENCODING A BETA-GLUCOSIDASE

被引:40
作者
LIN, LL [1 ]
RUMBAK, E [1 ]
ZAPPE, H [1 ]
THOMSON, JA [1 ]
WOODS, DR [1 ]
机构
[1] UNIV CAPE TOWN,DEPT MICROBIOL,RONDEBOSCH 7700,SOUTH AFRICA
来源
JOURNAL OF GENERAL MICROBIOLOGY | 1990年 / 136卷
关键词
D O I
10.1099/00221287-136-8-1567
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The cloning, expression and nucleotide sequence of a 3.74 kb DNA segment on pLS215 containing a β-glucosidase gene (bglA) from Butyrivibrio fibrisolvens H17c was investigated. The B. fibrisolvens bglA open reading frame (ORF) of 2490 bp encoded a β-glucosidase of 830 amino acid residues with a calculated M(r) of 91.800. In Escherichia coli C600(pLS215) cells the β-glucosidase was localized in the cytoplasm and these cells produced an additional protein with an apparent M(r) of approximately 94.000. The bglA gene was expressed from its own regulatory region in E. coli and a single mRNA initiation point was identified upstream of the bglA ORF and adjacent to a promoter consensus sequence. The primary structure of the β-glucosidase showed > 40% similarity with a domain of 237 amino acids present in the β-glucosidases of Kluyveromyces fragilis and Clostridium thermocellum. The B. fibrisolvens β-glucosidase hydrolysed cellobiose to a limited extent, cellotriose to cellobiose and glucose, and cellotetraose and cellopentaose to predominantly glucose.
引用
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页码:1567 / 1576
页数:10
相关论文
共 41 条
[1]  
AIBA H, 1981, J BIOL CHEM, V256, P1905
[2]   ISOLATION AND STRUCTURE OF A TRYPTIC GLYCOPEPTIDE FROM THE ACTIVE-SITE OF BETA-GLUCOSIDASE A3 FROM ASPERGILLUS-WENTII [J].
BAUSE, E ;
LEGLER, G .
BIOCHIMICA ET BIOPHYSICA ACTA, 1980, 626 (02) :459-465
[3]   CLONING AND SEQUENCING OF AN ENDOGLUCANASE (END1) GENE FROM BUTYRIVIBRIO-FIBRISOLVENS H17C [J].
BERGER, E ;
JONES, WA ;
JONES, DT ;
WOODS, DR .
MOLECULAR & GENERAL GENETICS, 1989, 219 (1-2) :193-198
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   MEDIUM WITHOUT RUMEN FLUID FOR NONSELECTIVE ENUMERATION AND ISOLATION OF RUMEN BACTERIA [J].
CALDWELL, DR ;
BRYANT, MP .
APPLIED MICROBIOLOGY, 1966, 14 (05) :794-&
[6]   POSITIVE-SELECTION CLONING VEHICLE USEFUL FOR OVERPRODUCTION OF HYBRID PROTEINS [J].
CHENG, SC ;
MODRICH, P .
JOURNAL OF BACTERIOLOGY, 1983, 154 (02) :1005-1008
[7]   THE PROPERTIES OF FUNGAL AND BACTERIAL CELLULASES WITH COMMENT ON THEIR PRODUCTION AND APPLICATION [J].
COUGHLAN, MP .
BIOTECHNOLOGY & GENETIC ENGINEERING REVIEWS, 1985, 3 :39-109
[8]   CONSENSUS SEQUENCE FOR ESCHERICHIA-COLI HEAT-SHOCK GENE PROMOTERS [J].
COWING, DW ;
BARDWELL, JCA ;
CRAIG, EA ;
WOOLFORD, C ;
HENDRIX, RW ;
GROSS, CA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (09) :2679-2683
[9]   PROLONGED INCUBATION IN CALCIUM-CHLORIDE IMPROVES THE COMPETENCE OF ESCHERICHIA-COLI-CELLS [J].
DAGERT, M ;
EHRLICH, SD .
GENE, 1979, 6 (01) :23-28
[10]   CHARACTERIZATION OF SEVERAL BOVINE RUMEN BACTERIA ISOLATED WITH A XYLAN MEDIUM [J].
DEHORITY, BA .
JOURNAL OF BACTERIOLOGY, 1966, 91 (05) :1724-&