Low‐pH production of <scp>d</scp>‐lactic acid using newly isolated acid tolerant yeast <i>Pichia kudriavzevii</i> NG7

Park, Hyun Joo; Bae, Jung Hoon; Ko, Hyeok Jin; Lee, Sun Hee; Sung, Bong Hyun; Han, Jong‐In; Sohn, Jae Hak

doi:10.1002/bit.26745

Cited by 73 publications

(78 citation statements)

References 33 publications

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“…It could thus be interesting to metabolically engineer a PDLA producing strain to produce its own D-lactic acid or, conversely, to engineer metabolic pathway for PDLA in an eukaryotic chassis capable of D-lactic acid production. In this context, D-lactic acid eukaryotic producer such as S. cerevisiae, Pichia kudriavzevii or Rhizopus oryzae (Sauer et al, 2010;Baek et al, 2016;Park et al, 2018) could be good candidates. However, biological polymer production raise the question of its extraction, as the purification of microbial biopolymers is one of the major challenges to overcome for their industrial development (Kosseva and Rusbandi, 2018).…”

Section: Discussionmentioning

confidence: 99%

Engineering the Yeast Yarrowia lipolytica for Production of Polylactic Acid Homopolymer

Lajus

Dusséaux

Verbeke

et al. 2020

Front. Bioeng. Biotechnol.

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Polylactic acid is a plastic polymer widely used in different applications from printing filaments for 3D printer to mulching films in agriculture, packaging materials, etc. Here, we report the production of poly-D-lactic acid (PDLA) in an engineered yeast strain of Yarrowia lipolytica. Firstly, the pathway for lactic acid consumption in this yeast was identified and interrupted. Then, the heterologous pathway for PDLA production, which contains a propionyl-CoA transferase (PCT) converting lactic acid into lactyl-CoA, and an evolved polyhydroxyalkanoic acid (PHA) synthase polymerizing lactyl-CoA, was introduced into the engineered strain. Among the different PCT proteins that were expressed in Y. lipolytica, the Clostridium propionicum PCT exhibited the highest efficiency in conversion of D-lactic acid to D-lactyl-CoA. We further evaluated the lactyl-CoA and PDLA productions by expressing this PCT and a variant of Pseudomonas aeruginosa PHA synthase at different subcellular localizations. The best PDLA production was obtained by expressing the PCT in the cytosol and the variant of PHA synthase in peroxisome. PDLA homopolymer accumulation in the cell reached 26 mg/g-DCW, and the molecular weights of the polymer (Mw = 50.5 × 10 3 g/mol and Mn = 12.5 × 10 3 g/mol) were among the highest reported for an in vivo production.

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Section: Discussionmentioning

confidence: 99%

Engineering the Yeast Yarrowia lipolytica for Production of Polylactic Acid Homopolymer

Lajus

Dusséaux

Verbeke

et al. 2020

Front. Bioeng. Biotechnol.

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“…Furthermore, recombinant I. orientalis strains can produce 15–20 g/L lactic acid under anaerobic conditions in an unbuffered medium at a pH of 2 ( Suominen et al., 2012 ) and succinic acid with a titer of 11.6 g/L ( Xiao et al., 2014 ). A recent isolate that was subsequently genetically modified and adapted to high lactic acid concentrations was reported to produce as much as 154 g/L d -lactic acid at a pH of 4.7 with the addition of calcium hydroxide ( Park et al., 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Genome-scale metabolic reconstruction of the non-model yeast Issatchenkia orientalis SD108 and its application to organic acids production

Suthers

Dinh

Fatma

et al. 2020

Metabolic Engineering Communications

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Many platform chemicals can be produced from renewable biomass by microorganisms, with organic acids making up a large fraction. Intolerance to the resulting low pH growth conditions, however, remains a challenge for the industrial production of organic acids by microorganisms. Issatchenkia orientalis SD108 is a promising host for industrial production because it is tolerant to acidic conditions as low as pH 2.0. With the goal to systematically assess the metabolic capabilities of this non-model yeast, we developed a genome-scale metabolic model for I. orientalis SD108 spanning 850 genes, 1826 reactions, and 1702 metabolites. In order to improve the model’s quantitative predictions, organism-specific macromolecular composition and ATP maintenance requirements were determined experimentally and implemented. We examined its network topology, including essential genes and flux coupling analysis and drew comparisons with the Yeast 8.3 model for Saccharomyces cerevisiae . We explored the carbon substrate utilization and examined the organism’s production potential for the industrially-relevant succinic acid, making use of the OptKnock framework to identify gene knockouts which couple production of the targeted chemical to biomass production. The genome-scale metabolic model iIsor 850 is a data-supported curated model which can inform genetic interventions for overproduction.

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“…It has been used for low-pH ethanol fermentation and engineered to produce lactic acid and succinic acid. 7,77,78 The organism natively favors HR for DNA repair, and recently an episomal plasmid as well as a CRISPR/ Cas9-based gene-editing tool have been reported. 55 For CRISPR/Cas9-based gene editing in I. orientalis, SpCas9 was expressed from an ARS plasmid using pTEF1 and the gRNA was expressed from the same plasmid, which also contained a homology donor with 50 bp arms containing an 8-bp frame shift mutation for the targeted gene.…”

Section: Issatchenkia Orientalismentioning

confidence: 99%

Recent advances in domesticating non‐model microorganisms

Fatma

Schultz

Zhao

2020

Biotechnology Progress

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Non-model microorganisms have been increasingly explored as microbial cell factories for production of chemicals, fuels, and materials owing to their unique physiology and metabolic capabilities. However, these microorganisms often lack facile genetic tools for strain development, which hinders their adoption as production hosts. In this review, we describe recent advances in domestication of non-model microorganisms, including bacteria, actinobacteria, cyanobacteria, yeast, and fungi, with a focus on the development of genetic tools. In addition, we highlight some successful applications of non-model microorganisms as microbial cell factories.

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Low‐pH production of d‐lactic acid using newly isolated acid tolerant yeast Pichia kudriavzevii NG7

Cited by 73 publications

References 33 publications

Engineering the Yeast Yarrowia lipolytica for Production of Polylactic Acid Homopolymer

Engineering the Yeast Yarrowia lipolytica for Production of Polylactic Acid Homopolymer

Genome-scale metabolic reconstruction of the non-model yeast Issatchenkia orientalis SD108 and its application to organic acids production

Recent advances in domesticating non‐model microorganisms

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