The combination of ultraviolet mutagenesis and PPX1 overexpression synergistically enhanced S-adenosyl-L-methionine synthesis in industrial Saccharomyces cerevisiae

Abstract

S-adenosyl-L-methionine (SAM) is the only injectable drug among the hepatoprotective and choleretic drugs, which has remarkable efficacy and is favored by hepatopaths. The demand for SAM is constantly increasing in clinical settings. Therefore, many efforts have been made to increase SAM biosynthesis from L-methionine and ATP in Saccharomyces cerevisiae. This study aimed to construct a stable and high-accumulating SAM industrial strain through successive ultraviolet irradiation (UV) mutations coupled with three resistant (ethionine, nystatin, and cordycepin, respectively) screening procedures and metabolic engineering strategies. Following multiple UV mutagenesis, a higher production mutant strain ZJT15–33 was successfully obtained. In addition, the recombinant strain spe2△-PPX1 was derived from ZJT15–33 by deleting the SPE2 and overexpressing the PPX1, resulting in a 2.5-fold enhanced ATP accumulation, which promoted the synthesis of 2.41 g/L SAM in the shake-flask, representing an 11.4-fold enhancement over the original strain (0.21 g/L). Furthermore, 11.65 g/L SAM was accumulated with 113 mg/g DCW SAM content in a 5-L fermenter at 96 h, marking a 36.57 % increase compared to strain ZJT15–33 (8.53 g/L). These results indicated that UV mutagenesis combined with PPX1 overexpression could effectively improve SAM synthesis in S. cerevisiae, providing a feasible approach for developing highly SAM industrial production.