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Gene cloning and expression of a detergent stable alkaline protease from Aspergillus clavatus ES1

Paper ID Volume ID Publish Year Pages File Format Full-Text
35521 45093 2010 7 PDF Available
Title
Gene cloning and expression of a detergent stable alkaline protease from Aspergillus clavatus ES1
Abstract

The genes, cDNA alpES1 and alpES1, encoding Aspergillus clavatus ES1 alkaline protease were amplified from complementary DNA (cDNA) and genomic DNA, respectively, cloned in pCR®II-TOPO plasmid and then sequenced. Sequence analysis of the cDNA alpES1 gene revealed an open reading frame (ORF) of 1212 bp encoding a pre–pro-protein of 403 amino acid residues consisting of a 21-aa signal peptide, a 100-aa pro-peptide and a 282-aa mature protein with a calculated molecular weight of 28.5 kDa. Compared to the cDNA alpES1 gene, the alpES1 gene contained three introns, which had 53, 57 and 54 bp, respectively. The cDNA alpES1 gene was then sub-cloned in pET-30b(+) and expressed in Escherichia coli BL21 (λDE3). The purified recombinant protease had a molecular weight of about 32 kDa estimated by SDS-PAGE. Kinetic parameters, Km and kcat values of the recombinant AlpES1 for casein, were 0.23 mM and 12.38 min−1, respectively. The catalytic efficiency (kcat/Km) was 53.82 min−1 mM−1.

Keywords
Alkaline protease; Aspergillus clavatus ES1; Gene cloning; Expression; Detergent
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Gene cloning and expression of a detergent stable alkaline protease from Aspergillus clavatus ES1
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 45, Issue 10, October 2010, Pages 1746–1752
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