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Biotransformation of β-Hydroxypyruvate and Glycolaldehyde to L-Erythrulose by Pichia pastoris strain GS115 overexpressing native Transketolase

Wei, Y-C; Braun-Galleani, S; Henriquez, M-J; Bandara, S; Nesbeth, DN; (2018) Biotransformation of β-Hydroxypyruvate and Glycolaldehyde to L-Erythrulose by Pichia pastoris strain GS115 overexpressing native Transketolase. Biotechnology Progress , 34 (1) pp. 99-106. 10.1002/btpr.2577. Green open access

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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 catalyse 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 methanolinducible 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 300µL reaction, TK150 samples from a 1L fed-batch fermentation achieved a maximum Lerythrulose space time yield (STY) of 46.58 g L -1 hr-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.

Type: Article
Title: Biotransformation of β-Hydroxypyruvate and Glycolaldehyde to L-Erythrulose by Pichia pastoris strain GS115 overexpressing native Transketolase
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/btpr.2577
Publisher version: http://dx.doi.org/10.1002/btpr.2577
Language: English
Additional information: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: Pichia pastoris; transketolase; whole cell biocatalyst; L-erythrulose; product inhibition
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10026227
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