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Overexpression of Candida rugosa lipase Lip1 via combined strategies in Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
16904 42621 2016 10 PDF Available
Title
Overexpression of Candida rugosa lipase Lip1 via combined strategies in Pichia pastoris
Abstract

•Codon-optimized CRL1 gene was overexpressed via combining strategy.•A 5 copies CRL1 gene of recombinant was conducted.•CRL1 co-expressed with two chaperones significantly enhanced expression.•The best recombinant strains were fermented in optimized conditions.

In this study, combined strategies were employed to heterologously overexpress Candida rugosa lipase Lip1 (CRL1) in a Pichia pastoris system. The LIP1 gene was systematically codon-optimized and synthesized in vitro. The Lip1 activity of a recombinant strain harboring three copies of the codon-optimized LIP1 gene reached 1200 U/mL in a shake flask culture. Higher lipase activity, 1450 U/mL, was obtained using a five copy number construct. Co-expressing one copy of the ERO1p and BiP chaperones with Lip1p, the CRL1 lipase yield further reached 1758 U/mL, which was significantly higher than that achieved by expressing Lip1p alone or only co-expressing one molecular chaperone. When cultivated in a 3 L fermenter under optimal conditions, the recombinant strain GS115/87-ZA-ERO1p-BiP #7, expressing the molecular chaperones Ero1p and BiP, produced 13,490 U/mL of lipase activity at 130 h, which was greater than the 11,400 U/mL of activity for the recombinant strain GS115/pAO815-α-mCRL1 #87, which did not express a molecular chaperone. This study indicates that a strategy of combining codon optimization with co-expression of molecular chaperones has great potential for the industrial-scale production of pure CRL1.

Keywords
Pichia pastoris; Candida rugosa lipase; Molecular chaperone; Codon optimization; Fed-batch fermentation
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Overexpression of Candida rugosa lipase Lip1 via combined strategies in Pichia pastoris
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 82, January 2016, Pages 115–124
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