Prunasin production using engineered Escherichia coli expressing UGT85A47 from Japanese apricot and UDP-glucose biosynthetic enzyme genes

• Published in: Bioscience, biotechnology, and biochemistry Vol. 82; no. 11; pp. 2021 - 2029
• Main Authors: Yamaguchi, Takuya, Asano, Yasuhisa
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 02.11.2018; Oxford University Press
• Subjects: Acetonitriles - metabolism; Biotransformation; Catalysis; Cloning, Molecular; Cyanogenic glycosides; Escherichia coli - genetics; Glucosyltransferases - genetics; Glucosyltransferases - metabolism; Hydrogen-Ion Concentration; Japanese apricot; metabolic engineering; microbial production; Nitriles - metabolism; Prunus armeniaca - enzymology; Temperature; UDP-glucosyl transferase; Uridine Diphosphate Glucose - biosynthesis; Uridine Diphosphate Glucose - metabolism; UDP-glucosyl transferase; microbial production; Japanese apricot; metabolic engineering; Cyanogenic glycosides
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1080/09168451.2018.1497942

Japanese apricot, Prunus mume Sieb. et Zucc., biosynthesizes the l-phenylalanine-derived cyanogenic glucosides prunasin and amygdalin. Prunasin has biological properties such as anti-inflammation, but plant extraction and chemical synthesis are impractical. In this study, we identified and characterized UGT85A47 from Japanese apricot. Further, UGT85A47 was utilized for prunasin microbial production. Full-length cDNA encoding UGT85A47 was isolated from Japanese apricot after 5ʹ- and 3ʹ-RACE. Recombinant UGT85A47 stoichiometrically catalyzed UDP-glucose consumption and synthesis of prunasin and UDP from mandelonitrile. Escherichia coli C41(DE3) cells expressing UGT85A47 produced prunasin (0.64 g/L) from racemic mandelonitrile and glucose. In addition, co-expression of genes encoding UDP-glucose biosynthetic enzymes (phosphoglucomutase and UTP-glucose 1-phosphate uridiltransferase) and polyphosphate kinase clearly improved prunasin production up to 2.3 g/L. These results showed that our whole-cell biocatalytic system is significantly more efficient than the existing prunasin production systems, such as chemical synthesis. Prunasin production using engineered Escherichia coli expressing UGT85A47 from Japanese apricot.