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Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda

Paper ID Volume ID Publish Year Pages File Format Full-Text
3323 164 2013 6 PDF Available
Title
Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda
Abstract

Klebsiella pneumoniae XM-4 is first isolated from a bacterial consortium. According to the 16S rRNA sequence, it has only 75% sequence similarity to Cellulomonas sp. However, a 999-bp open reading frame of K. pneumoniae, which encodes a putative endo-glucanase of 333 amino acids belonging to GH8 family and designated as Cel8A, is found closely to Cellulomonas uda with 95% similarity in amino acid sequence. This implies that the biological evolution between both strains has occurred. The cel8A is constructed in vectors of pET-22b(+) and pET-32a(+) with or without containing a signal peptide, and then cloned and expressed in Escherichia coli. The best endo-glucanase production of 25.4 mg/l is then obtainable by applying pET-32a(+) without the signal peptide. The recombinant CelA8 has an optimal specific activity of 62.4 U/mg against CMC or 23.3 U/mg against β-d-glucan at 55 °C and pH 5.0, indicating good potential for the industrial application.

► Cel8A from Klebsiella pneumoniae XM-4 has firstly been cloned and expressed in Escherichia coli. ► The highest recombinant protein was obtained by used pET-32a(+) without signal peptide to cel8A. ► 95% similarity of Cel8A to Cellulomonas uda showed the biological evolution between both strains.

Keywords
Klebsiella pneumoniae; Cellulomonas uda; Endo-glucanase; Signal peptide; Recombinant expression; Biological evolution
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Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 78, 15 September 2013, Pages 53–58
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us