Engineering of Biosynthesis Pathway and NADPH Supply for Improved L-5-Methyltetrahydrofolate Production by Lactococcus lactis

Lu Chuanchuan; Liu Yanfeng; Li Jianghua; Liu Long; Du Guocheng

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자료유형: 학술저널

저자정보: Lu Chuanchuan (Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of Education Jiangnan Universit) Liu Yanfeng (Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of Education Jiangnan Universit) Li Jianghua (Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of Education Jiangnan Universit) Liu Long (Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of Education Jiangnan Universit) Du Guocheng (Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of Education Jiangnan Universit)

저널정보: 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제31권 제1호

발행연도: 2021.1

수록면: 154 - 162 (9page)

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L-5-methyltetrahydrofolate (5-MTHF) is one of the biological active forms of folate, which is widely used as a nutraceutical. However, low yield and serious pollution associated with the chemical synthesis of 5-MTHF hampers its sustainable supply. In this study, 5-MTHF production was improved by engineering the 5-MTHF biosynthesis pathway and NADPH supply in Lactococcus lactis for developing a green and sustainable biosynthesis approach. Specifically, overexpressing the key rate-limiting enzyme methylenetetrahydrofolate reductase led to intracellular 5-MTHF accumulation, reaching 18 μg/l. Next, 5-MTHF synthesis was further enhanced by combinatorial overexpression of 5-MTHF synthesis pathway enzymes with methylenetetrahydrofolate reductase, resulting in 1.7- fold enhancement. The folate supply pathway was strengthened by expressing folE encoding GTP cyclohydrolase I, which increased 5-MTHF production 2.4-fold to 72 μg/l. Furthermore, glucose-6- phosphate dehydrogenase was overexpressed to improve the redox cofactor NADPH supply for 5- MTHF biosynthesis, which led to a 60% increase in intracellular NADPH and a 35% increase in 5- MTHF production (97 μg/l). To reduce formation of the by-product 5-formyltetrahydrofolate, overexpression of 5-formyltetrahydrofolate cyclo-ligase converted 5-formyltetrahydrofolate to 5,10-methyltetrahydrofolate, which enhanced the 5-MTHF titer to 132 μg/l. Finally, combinatorial addition of folate precursors to the fermentation medium boosted 5-MTHF production, reaching 300 μg/l. To the best of our knowledge, this titer is the highest achieved by L. lactis. This study lays the foundation for further engineering of L. lactis for efficient 5-MTHF biosynthesis.

#L-5-Methyltetrahydrofolate #Lactococcus lactis #metabolic engineering #strengthening synthetic pathway #NADPH supply

참고문헌 (47)

참고문헌 신청

Arcot J / 2005 / Folate : methods of analysis / Trends Food Sci. Technol 16:253~266

Asrar FM / 2005 / Bacterially synthesized folate and supplemental folic acid are absorbed across the large intestine of piglets / J. Nutr. Biochem 16:587~593

Kelly P / 1997 / Unmetabolized folic acid in serum : acute studies in subjects consuming fortified food and supplements / Am. Clin. Nutr 65:1790~1795

Bailey LB / 1999 / Folate metabolism and requirements / J. Nutr 129:779~782

Kim Y-I / 2000 / Methylenetetrahydrofolate reductase polymorphisms, folate, and cancer risk : a paradigm of gene-nutrient interactions in carcinogenesis / Nutr. Rev 58:205~209

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