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Elevated intracellular acetyl-CoA availability by acs2 overexpression and mls1 deletion combined with metK1 introduction enhanced SAM accumulation in Saccharomyces cerevisiae

Paper ID Volume ID Publish Year Pages File Format Full-Text
2772 131 2016 9 PDF Available
Title
Elevated intracellular acetyl-CoA availability by acs2 overexpression and mls1 deletion combined with metK1 introduction enhanced SAM accumulation in Saccharomyces cerevisiae
Abstract

•Over-expressing acs2 and deleting mls1 elevated cytosolic acetyl-CoA and Met levels.•Introducing metK1 from L. infantum caused a higher SAM synthase activity in yeast.•Over-expressing acs2 and metK1 and deleting mls1 notably enhanced SAM accumulation.

S-Adenosyl-l-methionine (SAM), with diverse pharmaceutical applications, is biosynthesized from l-methionine and ATP. To enhance SAM accumulation in Saccharomyces cerevisiae CGMCC 2842 (2842), a new strategy based on yeast acetyl-CoA metabolism combined with introducing a methionine adenosyltransferase (metK1) from Leishmania infantum, was presented here. It was found that over-expressing acs2 (encoding acetyl-CoA synthase) and deleting mls1 (encoding malate synthase) increased SAM by 0.86- and 1.30-fold, respectively. To eliminate feedback inhibition of SAM synthase, a codon-optimized metK1 was introduced into 2842, and an increase of 1.45-fold of SAM was observed. Subsequently, metK1 and acs2 were co-expressed in the mls1 deleted strain, obtained the highly SAM-productive strain Ymls1 △GAPmK, and 2.22 g/L of SAM accumulated, which was 3.36-fold that in 2842. Moreover, the Ymls1 △GAPmK strain yielded 6.06 g/L SAM, which was 9.18-fold that in 2842, by fed-batch fermentation in a 10-L fermenter. Finally, the isolation and purification of SAM from yeast cell and preparation of SAM sulfate were preliminarily investigated. This study demonstrated that up-regulating acs2 and deleting mls1, which elevated intracellular acetyl-CoA levels, effectively enhanced the intracellular methionine biosynthesis. The elevated intracellular acetyl-CoA levels ultimately enhanced SAM accumulation, whereas the introduction of metK1 enhanced the redirection of acetyl-CoA to SAM biosynthesis in Ymls1 △GAPmK strain.

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Keywords
S-Adenosyl-l-methionine; Biosynthesis; Intracellular acetyl-CoA levels; Recombinant DNA; Yeast; Metabolite over production
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Elevated intracellular acetyl-CoA availability by acs2 overexpression and mls1 deletion combined with metK1 introduction enhanced SAM accumulation in Saccharomyces cerevisiae
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 107, 15 March 2016, Pages 26–34
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