Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1, 3, 4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase. Issue 8 (August 2019)

• Record Type: Journal Article
• Authors: Braun-Galleani, Stephanie; Henríquez, Maria-José; Nesbeth, Darren N.
• Is Part Of: Heliyon. Volume 5:Issue 8(2019)
• Publication Date: 2019-08
• Subjects: Bioengineering -- Biotechnology -- Chemical engineering -- Komagataella phaffii -- Pichia pastoris -- Transaminase -- Whole cell biocatalyst; Research -- Periodicals; Medical sciences -- Periodicals; Natural history -- Periodicals; Social sciences -- Periodicals; Earth sciences -- Periodicals; Physical sciences -- Periodicals; 507.2
• DOI: 10.1016/j.heliyon.2019.e02338
• Languages: English
• ISSNs: 2405-8440

Abstract

Abstract: We have engineered strain BG-10 of the methylotrophic yeast Komagataella phaffii for use as an effective whole cell biocatalyst. We introduced into the yeast a transgene encoding a Chromobacterium violaceum ω-transaminase for transcription in response to methanol induction. The strain was then assessed with respect to its growth performance and biotransformation of a fed ketoalcohol substrate to an amino-alcohol. In the resultant strain, BG-TAM, methanol induction did not compromise cell growth. Successful bioconversion of fed substrates to the by-product, acetophenone, indicated transaminase activity in shake flask-cultivated BG-TAM cells. We then used bioreactor cultivation to exploit the high levels of biomass achievable by Komagataella phaffii . In a 900 μL reaction the BG-TAM strain at OD600 = 1024 achieved up to 0.41 mol mol −1 (molproduct molsubstrate −1 ) yield on substrate (Yp/s) for production of 1-methyl-3-phenylpropylamine and a space time yield (STY) of 0.29 g L −1 h −1 for production of 2-amino-1, 3, 4-butanetriol. We have shown that transamination, an important step for bespoke synthesis of small molecule medicines, is biologically realisable using enzymes with a broad substrate range, such as ω-transaminases, within living yeast cells that are fed low-cost substrates for bioconversion.