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Whole cell biosynthetic activity of Komagataella phaffii (Pichia pastoris) GS115 strains engineered with transgenes encoding Chromobacterium violaceum ω-transaminase alone or combined with native transketolase

Henríquez, M-J; Braun-Galleani, S; Nesbeth, D; (2020) Whole cell biosynthetic activity of Komagataella phaffii (Pichia pastoris) GS115 strains engineered with transgenes encoding Chromobacterium violaceum ω-transaminase alone or combined with native transketolase. Biotechnology Progress , 36 (1) , Article e2893. 10.1002/btpr.2893. Green open access

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Abstract

Whole cell biocatalysis is an ideal tool for biotransformations that demand enzyme regeneration or robustness to fluctuating pH, osmolarity and biocontaminant load in feedstocks. The methylotrophic yeast Komagataella phaffii is an attractive alternative to Escherichia coli for whole cell biocatalysis due to its genetic tractability and capacity to grow to up to 60% wet cell weight by volume. We sought to exploit high cell density K. phaffii to intensify whole‐cell chiral amino‐alcohol (CAA) biosynthesis. We engineered two novel K. phaffii GS115 strains: one by inserting a Chromobacterium violaceum ω‐transaminase CV2025 transgene, for strain PpTAmCV708, and a second strain, PpTAm‐TK16, by also inserting the same CV2025 transgene plus a second transgene for a native transketolase. At high cell density, both strains tolerated high substrate concentrations. When fed three low cost substrates, 200 mM glycolaldehyde, 200 mM hydroxypyruvate and 150 mM methylbenzylamine, PpTAm‐TK16 whole cells achieved 0.29 g L−1 hr−1 space–time yield of the acetophenone by‐product, a 49‐fold increase of the highest levels reported for E. coli whole cells harboring the equivalent pathway. When fed only the low‐cost substrate, 150 mM methylbenzylamine, strain PpTAmCV708 achieved a 105‐fold increase of reported E. coli whole cell biocatalysis performance, with a space–time yield of 0.62 g L−1 hr−1 of the CAA, 2‐amino‐1,3,4‐butanetriol (ABT). The rapid growth and high biomass characteristics of K. phaffii were successfully exploited for production of ABT by whole‐cell biocatalysis at higher levels than the previously achieved with E. coli in the presence of the same substrates.

Type: Article
Title: Whole cell biosynthetic activity of Komagataella phaffii (Pichia pastoris) GS115 strains engineered with transgenes encoding Chromobacterium violaceum ω-transaminase alone or combined with native transketolase
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/btpr.2893
Publisher version: https://doi.org/10.1002/btpr.2893
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: biocatalyst, chiral amino‐alcohol, Komagataella phaffii, Pichia pastoris, transaminase, transketolase, whole‐cell biocatalysis
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/10079441
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