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Biotransformation of β-hydroxypyruvate and Glycolaldehyde to L-Erythrulose by Pichia Pastoris Strain GS115 Overexpressing Native Transketolase

Biotransformation of β-hydroxypyruvate and Glycolaldehyde to L-Erythrulose by Pichia Pastoris Strain GS115 Overexpressing Native Transketolase

Yu-Chia Wei, Stephanie Braun-Galleani, Maria José Henríquez, Sahan Bandara, Darren Nesbeth

Biotechnol Prog. 2018 Jan;34(1):99-106.

PMID: 29086489

Abstract:

Transketolase is a proven biocatalytic tool for asymmetric carbon-carbon bond formation, both as a purified enzyme and within bacterial whole-cell biocatalysts. The performance of Pichia pastoris as a host for transketolase whole-cell biocatalysis was investigated using a transketolase-overexpressing strain to catalyze formation of l-erythrulose from β-hydroxypyruvic acid and glycolaldehyde substrates. Pichia pastoris transketolase coding sequence from the locus PAS_chr1-4_0150 was subcloned downstream of the methanol-inducible AOX1 promoter in a plasmid for transformation of strain GS115, generating strain TK150. Whole and disrupted TK150 cells from shake flasks achieved 62% and 65% conversion, respectively, under optimal pH and methanol induction conditions. In a 300 μL reaction, TK150 samples from a 1L fed-batch fermentation achieved a maximum l-erythrulose space time yield (STY) of 46.58 g L-1 h-1 , specific activity of 155 U gCDW-1, product yield on substrate (Yp/s ) of 0.52 mol mol-1 and product yield on catalyst (Yp/x ) of 2.23g gCDW-1. We have successfully exploited the rapid growth and high biomass characteristics of Pichia pastoris in whole cell biocatalysis. At high cell density, the engineered TK150 Pichia pastoris strain tolerated high concentrations of substrate and product to achieve high STY of the chiral sugar l-erythrulose. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:99-106, 2018.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP1113606	β-Hydroxypyruvic acid		1113-60-6	Price
AP533506	Erythrulose		533-50-6	Price

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