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Extracellular expression of YlLip11 with a native signal peptide from Yarrowia lipolytica MSR80 in three different yeast hosts

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논문

Extracellular expression of YlLip11 with a native signal peptide from Yarrowia lipolytica MSR80 in three different yeast hosts

학술지

Protein expression and purification

저자명

Kumari, Arti; Baronian, Keith; Kunze, Gotthard; Gupta, Rani

초록

<P><B>Abstract</B></P> <P>Lipase <I>YlLip11</I> from <I>Yarrowia lipolytica</I> was expressed with a signal peptide encoding sequence in <I>Arxula adeninivorans</I>, <I>Saccharomyces cerevisiae</I> and <I>Hansenula polymorpha</I> using the Xplor&reg;2 transformation/expression platform and an expression module with the constitutive <I>Arxula-</I>derived <I>TEF1</I> promoter. The YlLip11 signal peptide was functional in all of the yeast hosts with 97% of the recombinant enzyme being secreted into the culture medium. However, recombinant YlLip11 with His Tag fused at C-terminal was not active.</P> <P>The best recombinant YlLip11 producing <I>A. adeninivorans</I> G1212/YRC102-YlLip11 transformant cultivated in shake flasks produced 2654 U/L lipase, followed by <I>S. cerevisiae</I> SEY6210/YRC103-YlLip11 (1632U/L) and <I>H. polymorpha</I> RB11/YRC103-YlLip11 (1144U/L). Although the biochemical parameters of YlLip11 synthesized in different hosts were similar, their glycosylation level and thermo stability differed. The protein synthesized by the <I>H. polymorpha</I> transformant had the highest degree of glycosylation and with a <I>t</I> <SUB>1/2</SUB> of 60min at 70&deg;C, exhibited the highest thermostability.</P>

발행연도

2015

발행기관

Elsevier

ISSN

1046-5928

ISSN

1096-0279

110

페이지

pp.138-144

주제어

Lipase; Yarrowia lipolytica; Arxula adeninivorans; Saccharomyces cerevisiae; Hansenula polymorpha

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1 2023-12-11

논문; 2015-06-01

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