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Cloning, over-expression and characterization of an alkali-tolerant endo-β-1,4-mannanase from Penicillium freii F63

Paper ID Volume ID Publish Year Pages File Format Full-Text
21203 43211 2012 5 PDF Available
Title
Cloning, over-expression and characterization of an alkali-tolerant endo-β-1,4-mannanase from Penicillium freii F63
Abstract

A glycosyl hydrolase family 5 endo-β-mannanase gene (man5F63) was cloned from Penicillium freii F63 and overexpressed in Pichia pastoris. man5F63 contained an open reading frame of 1260 bp that encoded a polypeptide of 419 amino acids including a putative 18-residue signal peptide. The recombinant enzyme (rMan5F63) was secreted into the culture supernatant to near electrophoretic homogeneity with a high yield (1.1 g l–1 in flask). Its apparent molecular weight was approximately 72.0 kDa, 29.0 kDa higher than the theoretical molecular mass. rMan5F63 was optimal at pH 4.5 and 60°C and exhibited good stability over a broad pH range from acidic to alkaline (> 85.0% activity at pH 4.0–9.0, > 70.0% activity at pH 10.0 and 43.7% even at pH 12.0). The activity of rMan5F63 was significantly enhanced in the presence of Co2 +, Cu2 +, Mn2 + and β-mercaptoethanol and was strongly inhibited by Hg2 + and SDS. The specific activity, Km and Vmax values were 47.5 U mg− 1, 7.8 mg ml− 1 and 70.4 μmol min− 1 mg− 1, respectively, for locust bean gum, and 40.3 U mg− 1, 2.3 mg ml− 1 and 61.7 μmol min− 1 mg− 1, respectively, for konjac flour. All these favorable enzymatic properties make it cost-effective to commercialization and valuable in various industries.

Keywords
Penicillium freii F63; β-Mannanase; Pichia pastoris; Over-expression; Alkali-tolerant
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Cloning, over-expression and characterization of an alkali-tolerant endo-β-1,4-mannanase from Penicillium freii F63
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 113, Issue 6, June 2012, Pages 710–714
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