Development of Glutathione Production Technology Based on Designed Active Yeast Overproducers

TitleDevelopment of Glutathione Production Technology Based on Designed Active Yeast Overproducers
Publication TypeJournal Article
Year of Publication2015
AuthorsYurkiv, MT, Kurylenko, OO, Vasylyshyn, RV, Dmytruk, KV, Martynyuk, NB, Skorohod, VV, Sybirny, AA
Short TitleSci. innov.
DOI10.15407/scine11.05.053
Volume11
Issue5
SectionResearch and Engineering Innovative Projects of the National Academy of Sciences of Ukraine
Pagination53-60
LanguageEnglish
Abstract

The Hansenula polymorpha recombinant strain overexpressing both GSH2 gene encoding γ-glutamyl-cysteine-synthetase and MET4 gene encoding the transcription activator of genes involved in cysteine (precursor of glutathione) biosynthesis has been obtained using metabolic engineering approaches. The recombinant strain is characterized by significantly increased glutathione output as compared with in vitro wild-type strain. Conditions for efficient glutathione production by recombinant H. polymorpha strain have been optimized. A semi-industrial model for glutathione production using the designed H. polymorpha overproducer has been developed.

Keywordsand metabolic engineering., glutathione, Hansenula polymorpha, yeast
References: 

1. Penninckx M.J. A short review on the role of glutathione in the response of yeasts to nutritional, environmental, and oxidative stresses. Enzyme Microb Technol. 2000, 26: 737—742.
2. Pocsi I., Prade R., Penninckx M. Glutathione, Altruistic Metabolite in Fungi. Advances in Microbial Physiology. 2004, V. 49: 13—86.
3. Li Y., Wei G., Chen J. Glutatione: a review on biotechnological production. Applied Microbiology and Biotechnology. 2004: 305—315.
4. Gellisen G. Hansenula polymorpha — biology and applications. Weinheim: Wiley VCH; 2002.
5. Ubiyvovk V.M., Ananin V.M., Malyshev A.Y. et al. Оptimization of glutathione production in batch and fedbatch cultures by the wild-type and recombinant strains of the methylotrophic yeast Hansenula polymorpha DL-1. BMC Biotechnol. 2011, V.11: 8.
6. Sambrook J., Fritsh E.F., Maniatis T. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, New York. 1989.
7. Faber K.N., Haima P., Harder W. et al. Highly-efficient electrotransformation of the yeast Hansenula polymorpha. Curr Genet. 1994, 25: 305—310.
8. Griffith O., Mulcahy R. The enzyme of glutathione synthesis: γ-glutamylcysteine synthetase. Advances in enzymology and related aress of molecular biology. 1999, V. 73: 209—267.
9. Grabek-Lejko D, Kurylenko O.O, Sibirny V.A. et al. Alcoholic fermentation by wild-type Hansenula polymorpha and Saccharomyces cerevisiae versus recombinant strains with an elevated level of intracellular glutathione. J. Ind. Microbiol Biotechnol. 2011, 38(11): 1853-9.
http://dx.doi.org/10.1007/s10295-011-0974-z 
10. Taxis C., Knop M. System of centromeric, episomal, and integrative vectors based on drug resistance markers for Saccharomyces cerevisiae. BioTechniques. 2006, 40: 73—78.
11. Wang Z., Tan T., Song J. Effect of amino acids addition and feedback control strategies on the high-cell-density cultivation of Saccharomyces cerevisiae for glutathione production. Process Biochemistry. 2007, 42: 108—111.