Toward the construction of a technology platform for chemicals production from methanol: d-lactic acid production from methanol by an engineered yeast Pichia pastoris
- 53 Downloads
With the reduction in oil reserves and steady increases in the price of oil, alternative carbon sources like methanol are promising, but an efficient conversion process to fuels and other chemicals is still desired. In this study, we demonstrated for the first time the production of lactic acid from methanol using a lactate dehydrogenase copy number amplifying strategy in Pichia pastoris. We engineered methylotrophic yeast (Pichia pastoris) producing d-lactic acid by d-lactate dehydrogenase gene (d-LDH) integration into the non-transcribed spacer of the ribosomal DNA (rDNA) locus and post-transformational amplification. The resultant engineered strains GS115/S8/Z3 and GS115/S16/Z3 produced 3.48 and 3.26 g/L of d-lactic acid from methanol, respectively, in a 96-h test tube fermentation. To our knowledge, this is the first report about d-lactic acid production from methanol by an engineered P. pastoris strain. The technique of gene integration into the rDNA locus and post-transformational gene amplification could be useful for metabolic engineering in P. pastoris, and the chemical production from methanol by engineered P. pastoris represents a promising industrial technology.
Keywordsd-Lactic acid Fermentation Methanol Methylotrophic yeast Multi-copy integration Pichia pastoris
This work was partly supported by Japan Society for the Promotion of Science KAKENHI (Grant Number 18K14069) to RY.
Compliance with ethical standards
The authors declare that they have no conflict of interest.
- de Lima PB, Mulder KC, Melo NT, Carvalho LS, Menino GS, Mulinari E, de Castro VH, Dos Reis TF, Goldman GH, Magalhaes BS, Parachin NS (2016) Novel homologous lactate transporter improves l-lactic acid production from glycerol in recombinant strains of Pichia pastoris. Microb Cell Fact 15:158CrossRefGoogle Scholar
- Motoyama H, Anazawa H, Katsumata R, Araki K, Teshiba S (1993) Amino acid production from methanol by Methylobacillus glycogenes mutants: Isolation of l-glutamic acid hyper-producing mutants from M. glycogenes strains, and derivation of l-threonine and l-lysine-producing mutants from them. Biosci Biotechnol Biochem 57:82–87CrossRefGoogle Scholar
- Wendisch VF, Brito LF, Gil Lopez M, Hennig G, Pfeifenschneider J, Sgobba E, Veldmann KH (2016) The flexible feedstock concept in industrial biotechnology: Metabolic engineering of Escherichia coli, Corynebacterium glutamicum, Pseudomonas, Bacillus and yeast strains for access to alternative carbon sources. J Biotechnol 234:139–157CrossRefGoogle Scholar