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A synergistic effect on the production of S-adenosyl-l-methionine in Pichia pastoris by knocking in of S-adenosyl-l-methionine synthase and knocking out of cystathionine-β synthase

Paper ID Volume ID Publish Year Pages File Format Full-Text
25622 43585 2006 9 PDF Available
Title
A synergistic effect on the production of S-adenosyl-l-methionine in Pichia pastoris by knocking in of S-adenosyl-l-methionine synthase and knocking out of cystathionine-β synthase
Abstract

Two strategies of metabolic engineering have been used, individually or combined, to alter the metabolic flux to improve the production of S-adenosyl-l-methionine (SAM) in Pichia pastoris. One is over expressing SAM synthase by knock-in technique, the other is the disruption of cystathionine-β synthase (CBS) by knock-out technique. Strain Gsam with ectopic SAM synthase gene produced 20 times of SAM comparing to the starter strain GS115. Disruption of CBS in GS115 only doubled its SAM production. However, disruption of CBS in Gsam results in a robust increase of SAM production, more than 56 times of the strain GS115. Thus, we report for the first time a synergistic effect on the production of SAM in yeast by the combination of knock-in and knock-out techniques. Furthermore, we optimize the cultural conditions for the genetically modified strain Gsam-cbs to produce SAM. The maximum yield of SAM reaches 3.6 g/L in shake flask and 13.5 g/L in a 5 L fermentor, indicating that it could be used for industrial fermentation to produce large scale of SAM.

Keywords
S-adenosyl-l-methionine; Gene knock-out and knock-in; Metabolic engineering; Pichia pastoris
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A synergistic effect on the production of S-adenosyl-l-methionine in Pichia pastoris by knocking in of S-adenosyl-l-methionine synthase and knocking out of cystathionine-β synthase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 4, 1 December 2006, Pages 519–527
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