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Comparative performance of S-adenosyl-L-methionine biosynthesis and degradation in Pichia pastoris using different promoters and novel consumption inhibitors.

Abstract：

:The yeast Pichia pastoris is a widely used host for recombinant protein expression, and has recently been engineered for whole-cell biocatalysis. The inducible P(AOX) and constitutive P(GAP) promoters are commonly employed. In this study, the S-adenosyl-L-methionine (SAM) biosynthesis and degradation efficiency of two P. pastoris strains were compared, and novel inhibitors that suppress SAM degradation were characterized. The strains exhibited clear physiological differences. P(GAP)-Pichia showed higher transcription and activity of SAM synthetase, and the rapid cell growth led to higher levels of spermidine synthesis from SAM. In contrast, P(AOX)-Pichia synthesized higher levels of glutathione from SAM, and this strain responded to hydrogen peroxide formation during methanol utilization. Aristeromycin proved an efficient inhibitor of SAM degradation in P(AOX)-Pichia; 0.02 mg/L led to a 36.36% reduction in the ratio of glutathionine:SAM, and SAM accumulation was enhanced by 7.74% to 11.83 g/L. Ethanol was an even more efficient inhibitor of SAM consumption in P(GAP)-Pichia; 8 g/L resulted in a 73.68% decrease in the ratio of SPD:SAM, and SAM production was elevated by 54.55% to 0.17 g/L/h.

journal_name

Enzyme Microb Technol

journal_title

Enzyme and microbial technology

authors

Hu X,Chu J,Zhang S,Zhuang Y

doi

10.1016/j.enzmictec.2013.09.003

subject

Has Abstract

pub_date

2014-02-05 00:00:00

pages

94-9

eissn

0141-0229

issn

1879-0909

pii

S0141-0229(13)00192-0

journal_volume

55

pub_type

杂志文章

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