Document Detail

A cellulase gene from a new alkalophilic Bacillus sp. (strain N186-1). Its cloning, nucleotide sequence and expression in Escherichia coli.
MedLine Citation:
PMID:  8987645     Owner:  NLM     Status:  MEDLINE    
Several alkalophilic Bacillus spp. strains were selected for their capacity to produce alkaline cellulases. Culture supernatants of these strains showed optimal cellulase activities between pH 8 and 9 and they were stable from pH 6 to pH 12. A cellulase gene (celB1) from the alkalophilic Bacillus sp. strain N186-1 was cloned in Escherichia coli using polymerase chain reaction techniques. The cloned gene was present in a 2.539-bp HindIII fragment and its nucleotide sequence was determined. The coding sequence showed an open-reading frame encoding 389 amino acids. The amino acid sequence, deduced from the nucleotide sequence, permitted us to include it in family 5 (or A) of the glycosyl hydrolases. The complete open-reading frame of celB1 was cloned in the plasmid pET-11d and expressed in E. coli BL21 (DE3), in which a protein of 39 kDa was obtained in the cytoplasm; however, no endoglucanase activity was detected. A second construction in pET-12a allowed the production of a 39-kDa protein located in the periplasmic space of E. coli that had endoglucanase activity. The protein produced has optimal activity at pH 7 and 50 degrees C and it retains more than 70% of its activity after incubation for 1 h at pH 12.
J Sánchez-Torres; P Pérez; R I Santamaría
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Applied microbiology and biotechnology     Volume:  46     ISSN:  0175-7598     ISO Abbreviation:  Appl. Microbiol. Biotechnol.     Publication Date:  1996 Sep 
Date Detail:
Created Date:  1997-02-04     Completed Date:  1997-02-04     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8406612     Medline TA:  Appl Microbiol Biotechnol     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  149-55     Citation Subset:  B    
Departamento de Microbiología y Genética, Consejo Superior de Investigaciones Cientificas/Universidad de Salamanca, Spain.
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MeSH Terms
Amino Acid Sequence
Bacillus / enzymology*
Base Sequence
Cellulase / genetics*
Cloning, Molecular
DNA, Bacterial
Escherichia coli / genetics
Molecular Sequence Data
Open Reading Frames
Sequence Homology, Amino Acid
Reg. No./Substance:
0/DNA, Bacterial; EC

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