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Intracellular expression of Vitreoscilla hemoglobin improves production of Yarrowia lipolytica lipase LIP2 in a recombinant Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
17341 42661 2012 7 PDF Available
Title
Intracellular expression of Vitreoscilla hemoglobin improves production of Yarrowia lipolytica lipase LIP2 in a recombinant Pichia pastoris
Abstract

The Yarrowia lipolytica lipase LIP2 (YlLIP2) gene lip2 and Vitreoscilla hemoglobin gene vgb were co-expressed in Pichia pastoris, both under the control of AOX1 promoter, in order to alleviate respiration limitation under conditions of high cell-density fermentation and enhance YlLIP2 production. The results showed that recombinant P. pastoris strains harboring the lip2 and vgb genes (VHb+) displayed higher biomass and YlLIP2 activity than control strains (VHb−). Compared with VHb− cells, the expression levels of YlLIP2 in VHb-expressing cells when oxygen was not a limiting factor were improved 31.5% in shake-flask culture and 22% in a 10-L fermentor. Under non-limiting dissolved oxygen (DO) conditions, the maximum YlLIP2 activity of VHb+ in a 10-L fermentor reached 33,000 U/mL. Oxygen limitation had a more negative effect on YlLIP2 productivity in VHb− cells than in VHb+ cells. The highest YlLIP2 activity of VHb+ cells was approximately 1.84-fold higher than that of VHb− cells at lower DO levels. Moreover, the recombinant strain VHb+ exhibited a higher specific oxygen uptake rate and achieved higher cell viability under oxygen limiting and non-limiting conditions compared with VHb− cells. Therefore, the above results suggest that intracellular expression of VHb in recombinant P. pastoris has the potential to improve cell growth and industrial enzyme production.

► Lipase gene and Vitreoscilla hemoglobin gene were co-expressed in Pichia pastoris. ► Recombinant strain (VHb+) exhibited high specific oxygen uptake rate. ► VHb+ strain displayed higher biomass and lipase activity than control strains. ► The maximum lipase activity of VHb+ in a 10-L fermentor reached 33,000 U/mL. ► VHb+ strain also achieved higher cell viability than control strain.

Keywords
Expression; Lipase; Pichia pastoris; Vitreoscilla hemoglobin; Yarrowia lipolytica
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Intracellular expression of Vitreoscilla hemoglobin improves production of Yarrowia lipolytica lipase LIP2 in a recombinant Pichia pastoris
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 50, Issue 1, 5 January 2012, Pages 22–28
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