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Production of (R)-3-hydroxybutyric Acid by Arxula Adeninivorans

Production of (R)-3-hydroxybutyric Acid by Arxula Adeninivorans

Mateusz Biernacki, Jan Riechen, Urs Hähnel, Thomas Roick, Kim Baronian, Rüdiger Bode, Gotthard Kunze

AMB Express. 2017 Dec;7(1):4.

PMID: 28050847

Abstract:

(R)-3-hydroxybutyric acid can be used in industrial and health applications. The synthesis pathway comprises two enzymes, β-ketothiolase and acetoacetyl-CoA reductase which convert cytoplasmic acetyl-CoA to (R)-3-hydroxybutyric acid [(R)-3-HB] which is released into the culture medium. In the present study we used the non-conventional yeast, Arxula adeninivorans, for the synthesis enantiopure (R)-3-HB. To establish optimal production, we investigated three different endogenous yeast thiolases (Akat1p, Akat2p, Akat4p) and three bacterial thiolases (atoBp, thlp, phaAp) in combination with an enantiospecific reductase (phaBp) from Cupriavidus necator H16 and endogenous yeast reductases (Atpk2p, Afox2p). We found that Arxula is able to release (R)-3-HB used an existing secretion system negating the need to engineer membrane transport. Overexpression of thl and phaB genes in organisms cultured in a shaking flask resulted in 4.84 g L-1 (R)-3-HB, at a rate of 0.023 g L-1 h-1 over 214 h. Fed-batch culturing with glucose as a carbon source did not improve the yield, but a similar level was reached with a shorter incubation period [3.78 g L-1 of (R)-3-HB at 89 h] and the rate of production was doubled to 0.043 g L-1 h-1 which is higher than any levels in yeast reported to date. The secreted (R)-3-HB was 99.9% pure. This is the first evidence of enantiopure (R)-3-HB synthesis using yeast as a production host and glucose as a carbon source.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP625729	(R)-3-Hydroxybutyric acid		625-72-9	Price

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