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KMID : 1094720160210030408
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Biotechnology and Bioprocess Engineering
2016 Volume.21 No. 3 p.408 ~ p.413
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Expression system for Yarrowia lipolytica based on a promoter of the mitochondrial potential-dependent porin VDAC gene
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Epova Ekaterina Yu
Balovneva Maria V.
Isakova Elena P.
Kudykina Yuliya K.
Zylkova Marina V.
Deryabina Yulia I.
Shevelev Alexei B.
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Abstract
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In this study, we designed an expression system for the Y. lipolytica yeast, which can be fully efficient in media with non-standard industrial ingredients. Previously, we reported that the mitochondrial Voltage Dependent Anion Channel (VDAC) was a major protein overproduced in the Yarrowia lipolytica yeast in an alkaline (pH = 9.0) culture medium. In this study, the VDAC promoter was cloned and tested using a reporter system based on the LacZ gene. Naturally, the VDAC gene contains an intron located just within the ATG translation initiation codon. The VDAC promoter V2 variant with the intron and V3 variant without the intron were studied. The VDAC-driven clones exhibited high variability of the expression profile. Some clones were more active than the clones induced by the artificial hp4d promoter, when grown in both complete and low-cost industrial ingredient (10% fish and sunflower meal) containing media. The new expression system may greatly expand the range of recombinant producers of feed enzymes and some other types of fodder additives in the Y. lipolytica yeast, appropriate for assimilation of low-cost and non-standard raw material.
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KEYWORD
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expression, LacZ, vector, VDAC, Yarrowia lipolytica
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