2004•Journal of Zhejiang University Agriculture and Life SciencesRequires access
Cloning and Expression of β-1,3-1,4-glucanase Gene from Bacillus pumilus.
Jian-Yi Sun
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Abstract
A gene encoding β-glucanase from Bacillus pumilus has been cloned and expressed in Escherichia coli. The whole length of the gene was 754 bp, including an ORF of 717 bp encoding 239 amino acids. The enzyme showed a molecular size of 26.98 kD and a pI of 6.08.The gene showed a high degree of similarity to B.pumilus,amounting to homology of 91%. The gene was firstly registered in GenBank (Accession No: AY164456). The expression plasmid named pET-pum was constructed and transformed into E.coli BL21.The result of SDS-PAGE showed that a protein of about 27 kD was expressed in E.coli. The enzyme activity expressed in E.coli was 7.24 U/mL. The optimal pH and temperature for the this enzyme activity were respectively found to be about 5 and 50 ℃.