Construction of lactic acid-tolerant Saccharomyces cerevisiae by using CRISPR-Cas-mediated genome evolution for efficient d-lactic acid production

Mitsui, Ryosuke; Yamada, R.; Matsumoto, Takuya; Yoshihara, Shizue; Tokumoto, Hayato; Ogino, Hiroyasu

doi:10.1007/s00253-020-10906-3

Cited by 29 publications

(17 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The S. cerevisiae platform can be further developed for production of PHA polymers in non-buffered or low pH conditions (Fernandes et al., 2005 ; Mitsui et al., 2020 ; Rajkumar et al., 2016 ; Zhong et al. 2021 ), and for utilization of various industrial side streams as raw materials.…”

Section: Discussionmentioning

confidence: 99%

Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae

Ylinen

Maaheimo

Anghelescu-Hakala

et al. 2021

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

Polyhydroxyalkanoates (PHAs) provide biodegradable and bio-based alternatives to conventional plastics. Incorporation of 2-hydroxy acid monomers into polymer, in addition to 3-hydroxy acids, offers possibility to tailor the polymer properties. In this study, poly(D-lactic acid) (PDLA) and copolymer P(LA-3HB) were produced and characterized for the first time in the yeast Saccharomyces cerevisiae. Expression of engineered PHA synthase PhaC1437Ps6–19, propionyl-CoA transferase Pct540Cp, acetyl-CoA acetyltransferase PhaA, and acetoacetyl-CoA reductase PhaB1 resulted in accumulation of 3.6% P(LA-3HB) and expression of engineered enzymes PhaC1Pre and PctMe resulted in accumulation of 0.73% PDLA of the cell dry weight (CDW). According to NMR, P(LA-3HB) contained D-lactic acid repeating sequences. For reference, expression of PhaA, PhaB1, and PHA synthase PhaC1 resulted in accumulation 11% poly(hydroxybutyrate) (PHB) of the CDW. Weight average molecular weights of these polymers were comparable to similar polymers produced by bacterial strains, 24.6, 6.3, and 1 130 kDa for P(LA-3HB), PDLA, and PHB, respectively. The results suggest that yeast, as a robust and acid tolerant industrial production organism, could be suitable for production of 2-hydroxy acid containing PHAs from sugars or from 2-hydroxy acid containing raw materials. Moreover, the wide substrate specificity of PHA synthase enzymes employed increases the possibilities for modifying copolymer properties in yeast in the future.

show abstract

Section: Discussionmentioning

confidence: 99%

Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae

Ylinen

Maaheimo

Anghelescu-Hakala

et al. 2021

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

show abstract

“…In previous studies, ALE was also successfully applied to develop lactic acid-tolerant S. cerevisiae [17,18,42]. For the evolution conditions, lactic acid concentration gradually increased from 1% to 4% during 11 subcultures, and the evolved mutant showed no growth reduction with 1% lactic acid [17,18].…”

Section: Ale Of Lactic Acid-producing S Cerevisiae In a High Concentration Of Lactic Acidmentioning

confidence: 99%

“…For the evolution conditions, lactic acid concentration gradually increased from 1% to 4% during 11 subcultures, and the evolved mutant showed no growth reduction with 1% lactic acid [17,18]. In another study, evolution was performed under 8% lactic acid conditions, and the resulting strain showed a maximum tolerance to 6% lactic acid and not growth on 8% lactic acid [42]. In the present study, BK01 was able to grow on 8% lactic acid, which was a higher tolerance than the previously reported strains.…”

Section: Ale Of Lactic Acid-producing S Cerevisiae In a High Concentration Of Lactic Acidmentioning

confidence: 99%

l-Lactic Acid Production Using Engineered Saccharomyces cerevisiae with Improved Organic Acid Tolerance

Jang

Jeong

et al. 2021

JoF

View full text Add to dashboard Cite

Lactic acid is mainly used to produce bio-based, bio-degradable polylactic acid. For industrial production of lactic acid, engineered Saccharomyces cerevisiae can be used. To avoid cellular toxicity caused by lactic acid accumulation, pH-neutralizing agents are used, leading to increased production costs. In this study, lactic acid-producing S. cerevisiae BK01 was developed with improved lactic acid tolerance through adaptive laboratory evolution (ALE) on 8% lactic acid. The genetic basis of BK01 could not be determined, suggesting complex mechanisms associated with lactic acid tolerance. However, BK01 had distinctive metabolomic traits clearly separated from the parental strain, and lactic acid production was improved by 17% (from 102 g/L to 119 g/L). To the best of our knowledge, this is the highest lactic acid titer produced by engineered S. cerevisiae without the use of pH neutralizers. Moreover, cellulosic lactic acid production by BK01 was demonstrated using acetate-rich buckwheat husk hydrolysates. Particularly, BK01 revealed improved tolerance against acetic acid of the hydrolysates, a major fermentation inhibitor of lignocellulosic biomass. In short, ALE with a high concentration of lactic acid improved lactic acid production as well as acetic acid tolerance of BK01, suggesting a potential for economically viable cellulosic lactic acid production.

show abstract

“…As genome structural rearrangements can be used to generate strains with increased tolerance to stress, Mitsui et al further combined CRISPR-d integration aiming to obtain acid tolerance strain with rational metabolic engineering that integrates a series of genes from Leuconostoc mesenteroides into the yeast genome to enable the yeast strain to produce LA. 103 This combined method resulted in a high yield of D-LA.…”

Section: 1mentioning

confidence: 99%

Directed genome evolution driven by structural rearrangement techniques

Zhou

Xie

et al. 2021

Chem. Soc. Rev.

View full text Add to dashboard Cite

show abstract

Construction of lactic acid-tolerant Saccharomyces cerevisiae by using CRISPR-Cas-mediated genome evolution for efficient d-lactic acid production

Cited by 29 publications

References 30 publications

Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae

Production of D-lactic acid containing polyhydroxyalkanoate polymers in yeast Saccharomyces cerevisiae

l-Lactic Acid Production Using Engineered Saccharomyces cerevisiae with Improved Organic Acid Tolerance

Directed genome evolution driven by structural rearrangement techniques

Contact Info

Product

Resources

About