Effect of Pyruvate Decarboxylase Knockout on Product Distribution Using Pichia Pastoris (Komagataella Phaffii) Engineered for Lactic Acid Production

Overview

Journal Bioengineering (Basel)

Date 2018 Feb 22

PMID 29462904

Citations 8

Authors

Nadiele T M Melo

Kelly C L Mulder

Andre Moraes Nicola

Lucas S Carvalho

Gisele S Menino

Eduardo Mulinari

Nadia S Parachin

Affiliations

Soon will be listed here.

Abstract

Lactic acid is the monomer unit of the bioplastic poly-lactic acid (PLA). One candidate organism for lactic acid production is , a yeast widely used for heterologous protein production. Nevertheless, this yeast has a poor fermentative capability that can be modulated by controlling oxygen levels. In a previous study, lactate dehydrogenase (LDH) activity was introduced into enabling this yeast to produce lactic acid. The present study aimed to increase the flow of pyruvate towards the production of lactic acid in . To this end, a strain designated GLp was constructed by inserting the bovine lactic acid dehydrogenase gene (LDHb) concomitantly with the interruption of the gene encoding pyruvate decarboxylase (PDC). Aerobic fermentation, followed by micro-aerophilic culture two-phase fermentations, showed that the GLp strain achieved a lactic acid yield of 0.65 g/g. The distribution of fermentation products demonstrated that the acetate titer was reduced by 20% in the GLp strain with a concomitant increase in arabitol production: arabitol increased from 0.025 g/g to 0.174 g/g when compared to the GS115 strain. Taken together, the results show a significant potential for in producing lactic acid. Moreover, for the first time, physiological data regarding co-product formation have indicated the redox balance limitations of this yeast.

Citing Articles

Native and Recombinant Yeast Producers of Lactic Acid: Characteristics and Perspectives.

Tsaruk A, Filip K, Sibirny A, Ruchala J Int J Mol Sci. 2025; 26(5).

PMID: 40076630 PMC: 11900929. DOI: 10.3390/ijms26052007.

Engineering Komagataella phaffii for ethylene glycol production from xylose.

Carneiro C, Trichez D, Bergmann J, Reis V, Wagner N, Walther T AMB Express. 2024; 14(1):131.

PMID: 39614978 PMC: 11608209. DOI: 10.1186/s13568-024-01795-0.

Just around the Corner: Advances in the Optimization of Yeasts and Filamentous Fungi for Lactic Acid Production.

Melo N, de Oliveira Junqueira A, Lima L, Sampaio de Oliveira K, Dos Reis M, Franco O J Fungi (Basel). 2024; 10(3).

PMID: 38535215 PMC: 10971269. DOI: 10.3390/jof10030207.

Loss of a Functional Mitochondrial Pyruvate Carrier in Does Not Improve Lactic Acid Production from Glycerol in Aerobic Cultivation.

de Oliveira Junqueira A, Melo N, Skorupa Parachin N, Paes H Microorganisms. 2023; 11(2).

PMID: 36838448 PMC: 9967928. DOI: 10.3390/microorganisms11020483.

Transcriptome Profiling Analysis of Phosphate-Solubilizing Mechanism of Strain W134.

Wang S, Li Y, Zhang J, Wang X, Hong J, Qiu C Microorganisms. 2022; 10(10).

PMID: 36296274 PMC: 9609647. DOI: 10.3390/microorganisms10101998.

References

Sauer M, Porro D, Mattanovich D, Branduardi P . 16 years research on lactic acid production with yeast - ready for the market?. Biotechnol Genet Eng Rev. 2011; 27:229-56. DOI: 10.1080/02648725.2010.10648152. View

Li P, Anumanthan A, Gao X, Ilangovan K, Suzara V, Duzgunes N . Expression of recombinant proteins in Pichia pastoris. Appl Biochem Biotechnol. 2007; 142(2):105-24. DOI: 10.1007/s12010-007-0003-x. View

Branduardi P, Valli M, Brambilla L, Sauer M, Alberghina L, Porro D . The yeast Zygosaccharomyces bailii: a new host for heterologous protein production, secretion and for metabolic engineering applications. FEMS Yeast Res. 2004; 4(4-5):493-504. DOI: 10.1016/S1567-1356(03)00200-9. View

Lee J, Kang C, Lee S, Park Y, Cho K . Engineering cellular redox balance in Saccharomyces cerevisiae for improved production of L-lactic acid. Biotechnol Bioeng. 2014; 112(4):751-8. DOI: 10.1002/bit.25488. View

Tsuji H . Poly(lactic acid) stereocomplexes: A decade of progress. Adv Drug Deliv Rev. 2016; 107:97-135. DOI: 10.1016/j.addr.2016.04.017. View

Kuberl A, Schneider J, Thallinger G, Anderl I, Wibberg D, Hajek T . High-quality genome sequence of Pichia pastoris CBS7435. J Biotechnol. 2011; 154(4):312-20. DOI: 10.1016/j.jbiotec.2011.04.014. View

Kurtzman C . Description of Komagataella phaffii sp. nov. and the transfer of Pichia pseudopastoris to the methylotrophic yeast genus Komagataella. Int J Syst Evol Microbiol. 2005; 55(Pt 2):973-976. DOI: 10.1099/ijs.0.63491-0. View

Osawa F, Fujii T, Nishida T, Tada N, Ohnishi T, Kobayashi O . Efficient production of L-lactic acid by Crabtree-negative yeast Candida boidinii. Yeast. 2009; 26(9):485-96. DOI: 10.1002/yea.1702. View

Ilmen M, Koivuranta K, Ruohonen L, Rajgarhia V, Suominen P, Penttila M . Production of L-lactic acid by the yeast Candida sonorensis expressing heterologous bacterial and fungal lactate dehydrogenases. Microb Cell Fact. 2013; 12:53. PMC: 3680033. DOI: 10.1186/1475-2859-12-53. View

10.

Love K, Shah K, Whittaker C, Wu J, Bartlett M, Ma D . Comparative genomics and transcriptomics of Pichia pastoris. BMC Genomics. 2016; 17:550. PMC: 4974788. DOI: 10.1186/s12864-016-2876-y. View

11.

Hohmann S . Characterization of PDC6, a third structural gene for pyruvate decarboxylase in Saccharomyces cerevisiae. J Bacteriol. 1991; 173(24):7963-9. PMC: 212591. DOI: 10.1128/jb.173.24.7963-7969.1991. View

12.

Porro D, Brambilla L, Ranzi B, Martegani E, Alberghina L . Development of metabolically engineered Saccharomyces cerevisiae cells for the production of lactic acid. Biotechnol Prog. 1995; 11(3):294-8. DOI: 10.1021/bp00033a009. View

13.

Looser V, Bruhlmann B, Bumbak F, Stenger C, Costa M, Camattari A . Cultivation strategies to enhance productivity of Pichia pastoris: A review. Biotechnol Adv. 2015; 33(6 Pt 2):1177-93. DOI: 10.1016/j.biotechadv.2015.05.008. View

14.

Cheng H, Lv J, Wang H, Wang B, Li Z, Deng Z . Genetically engineered Pichia pastoris yeast for conversion of glucose to xylitol by a single-fermentation process. Appl Microbiol Biotechnol. 2014; 98(8):3539-52. DOI: 10.1007/s00253-013-5501-x. View

15.

Cregg J, Barringer K, Hessler A, Madden K . Pichia pastoris as a host system for transformations. Mol Cell Biol. 1985; 5(12):3376-85. PMC: 369166. DOI: 10.1128/mcb.5.12.3376-3385.1985. View

16.

Bianchi M, Brambilla L, Protani F, Liu C, Lievense J, Porro D . Efficient homolactic fermentation by Kluyveromyces lactis strains defective in pyruvate utilization and transformed with the heterologous LDH gene. Appl Environ Microbiol. 2001; 67(12):5621-5. PMC: 93352. DOI: 10.1128/AEM.67.12.5621-5625.2001. View

17.

Kurtzman C . Biotechnological strains of Komagataella (Pichia) pastoris are Komagataella phaffii as determined from multigene sequence analysis. J Ind Microbiol Biotechnol. 2009; 36(11):1435-8. DOI: 10.1007/s10295-009-0638-4. View

18.

Agarwal P, Uppada V, Noronha S . Comparison of pyruvate decarboxylases from Saccharomyces cerevisiae and Komagataella pastoris (Pichia pastoris). Appl Microbiol Biotechnol. 2013; 97(21):9439-49. DOI: 10.1007/s00253-013-4758-4. View

19.

Koganti S, Kuo T, Kurtzman C, Smith N, Ju L . Production of arabitol from glycerol: strain screening and study of factors affecting production yield. Appl Microbiol Biotechnol. 2010; 90(1):257-67. DOI: 10.1007/s00253-010-3015-3. View

20.

Porro D, Bianchi M, Brambilla L, Menghini R, Bolzani D, Carrera V . Replacement of a metabolic pathway for large-scale production of lactic acid from engineered yeasts. Appl Environ Microbiol. 1999; 65(9):4211-5. PMC: 99761. DOI: 10.1128/AEM.65.9.4211-4215.1999. View