Enhanced S-adenosyl-l-methionine production in Saccharomyces cerevisiae by spaceflight culture, overexpressing methionine adenosyltransferase and optimizing cultivation. 2012

Y Huang, and X Gou, and H Hu, and Q Xu, and Y Lu, and J Cheng
Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, China.

OBJECTIVE S-adenosyl-l-methionine (SAM) is an important biochemical molecule with great potential in the pharmacological and chemotherapeutic fields. In this study, our aims were to enhance SAM production in Saccharomyces cerevisiae. RESULTS Through spaceflight culture, a SAM-accumulating strain, S. cerevisiae H5M147, was isolated and found to produce 86·89% more SAM than its ground control strain H5. Amplified fragment length polymorphism (AFLP) analysis demonstrated that there were genetic variations between strain H5M147 and its ground control. Through recombinant DNA technology, the heterologous gene encoding methionine adenosyltransferase was integrated into the genome of strain H5M147. The recombinant strain H5MR83 was selected because its SAM production was increased by 42·98% when compared to strain H5M147. Furthermore, cultivation conditions were optimized using the one-factor-at-a-time and Taguchi methods. Under optimal conditions, strain H5MR83 yielded 7·76 g l(-1) of SAM in shake flask, an increase of 536·07% when compared to the strain H5. Furthermore, 9·64 g l(-1) of SAM was produced in fermenter cultivation. CONCLUSIONS A new SAM-accumulating strain, S. cerevisiae H5MR83, was obtained through spaceflight culture and genetic modification. Under optimal conditions, SAM production was increased to a relative high level in our study. CONCLUSIONS Through comprehensive application of multiple methods including spaceflight culture, genetic modification and optimizing cultivation, the yield of SAM could be increased by 6·4 times compared to that in the control strain H5. The obtained S. cerevisiae H5MR83 produced 7·76 g l(-1) of SAM in the flask cultures, a significant improvement on previously reported results. The SAM production period with S. cerevisiae H5MR83 was 84 h, which is shorter than previously reported results. Saccharomyces cerevisiae H5MR83 has considerable potential for use in industrial applications.

UI MeSH Term Description Entries
D008716 Methionine Adenosyltransferase An enzyme that catalyzes the synthesis of S-adenosylmethionine from methionine and ATP. EC 2.5.1.6. S-Adenosylmethionine Synthetase,ATP-Methionine S-Adenosyltransferase,ATP Methionine S Adenosyltransferase,Adenosyltransferase, Methionine,S Adenosylmethionine Synthetase,S-Adenosyltransferase, ATP-Methionine,Synthetase, S-Adenosylmethionine
D005818 Genetic Engineering Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc. Genetic Intervention,Engineering, Genetic,Intervention, Genetic,Genetic Interventions,Interventions, Genetic
D012436 S-Adenosylmethionine Physiologic methyl radical donor involved in enzymatic transmethylation reactions and present in all living organisms. It possesses anti-inflammatory activity and has been used in treatment of chronic liver disease. (From Merck, 11th ed) AdoMet,Ademetionine,FO-1561,Gumbaral,S Amet,S-Adenosyl-L-Methionine,S-Adenosylmethionine Sulfate Tosylate,SAM-e,Samyr,FO 1561,FO1561,S Adenosyl L Methionine,S Adenosylmethionine,S Adenosylmethionine Sulfate Tosylate
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D013026 Space Flight Travel beyond the earth's atmosphere. Space Exploration,Space Travel,Spaceflight,Exploration, Space,Explorations, Space,Flight, Space,Flights, Space,Space Explorations,Space Flights,Space Travels,Spaceflights,Travel, Space,Travels, Space
D054458 Amplified Fragment Length Polymorphism Analysis The detection of RESTRICTION FRAGMENT LENGTH POLYMORPHISMS by selective PCR amplification of restriction fragments derived from genomic DNA followed by electrophoretic analysis of the amplified restriction fragments. AFLP Analysis,AFLP Analyses,Analyses, AFLP,Analysis, AFLP
D019149 Bioreactors Tools or devices for generating products using the synthetic or chemical conversion capacity of a biological system. They can be classical fermentors, cell culture perfusion systems, or enzyme bioreactors. For production of proteins or enzymes, recombinant microorganisms such as bacteria, mammalian cells, or insect or plant cells are usually chosen. Fermentors,Bioreactor,Fermentor

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