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Comparison of the biochemical properties of a recombinant lipase extract from Rhizopus oryzae expressed in Pichia pastoris with a native extract

Paper ID Volume ID Publish Year Pages File Format Full-Text
3782 188 2011 7 PDF Available
Title
Comparison of the biochemical properties of a recombinant lipase extract from Rhizopus oryzae expressed in Pichia pastoris with a native extract
Abstract

Extracts containing mature Rhizopus oryzae lipase overexpressed in Pichia pastoris (rROL) and commercial ones from the native microorganism (nROL) have been characterized. The specific activity of rROL extract was more than 40-fold higher than nROL. The presence of multiple bands of lipases around 34 kDa was detected by western blot and zymogram analysis in both extracts. Nevertheless, rROL showed a slightly lower molecular weight than nROL. The presence of hydrolytic activity not recognised as derived from a lipase was also detected in nROL at higher molecular weights.The influence of the ionic strength on lipase activity was assayed and there was an effect both on pH and optimal temperature. Some differences were found between the two extracts. The specificity against triacylglycerol esters and p-nitrophenol substrates was also analyzed. Similar behaviour was noted towards triacylglycerol esters; however rROL showed opposite behaviour compared to nROL towards p-nitrophenol esters with preferences for long chain derivatives.The properties of rROL were partially different from those reported for nROL showing the influence of the pre-pro-sequence of ROL, the post-translational modifications of Pichia pastoris and the effect of an esterase in nROL powder.

Research highlights► Recombinant lipase has higher specific activity than native one. ► More than one form of lipase was found in both extracts. ► An esterase activity was found in the native extract. ► Different specificity towards p-nitrophenol esters. ► Determination of the influence of ionic strength on lipolytic activity.

Keywords
Pichia pastoris; Rhizopus oryzae; Recombinant lipase; Biochemical properties; Protein extract; Specificity
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Comparison of the biochemical properties of a recombinant lipase extract from Rhizopus oryzae expressed in Pichia pastoris with a native extract
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 54, Issue 2, 15 April 2011, Pages 117–123
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