Biotransformation of L-lysine to L-pipecolic acid catalyzed by L-lysine 6-aminotransferase and pyrroline-5-carboxylate reductase

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2002-03, Vol.66 (3), p.622-627
• Main Authors: Fujii, T. (Mercian Corp., Fujisawa, Kanagawa (Japan). Central Research Labs.), Mukaihara, M, Agematu, H, Tsunekawa, H
• Format: Article
• Language: English
• Subjects: AMINOTRANSFERASES; BIOCONVERSION; Bioconversions. Hemisynthesis; Biological and medical sciences; Biotechnology; Chromatography, High Pressure Liquid; Chromatography, Liquid; Chromatography, Thin Layer; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fermentation; Fundamental and applied biological sciences. Psychology; Genetic Complementation Test; L-Lysine 6-Transaminase; L-pipecolic acid; L-pipecolic acid (L-PA); lat; lat gene; LYSINE; Lysine - metabolism; lysine 6-aminotransferase; Mass Spectrometry; Methods. Procedures. Technologies; Oxidation-Reduction; OXIDOREDUCTASES; Pipecolic Acids - metabolism; piperideine-6-carboxylate (P6C); Plasmids - genetics; proC; proC gene; Pyrroline Carboxylate Reductases - genetics; Pyrroline Carboxylate Reductases - metabolism; pyrroline-5-carboxylate (P5C) reductase; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Transaminases - genetics; Transaminases - metabolism
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.66.622

The enzyme involved in the reduction of D1-piperideine-6-carboxylate (P6C) to L-pipecolic acid (L-PA) has never been identified. We found that Escherichia coli JM109 transformed with the lot gene encoding L-lysine 6-aminotransferase (LAT) converted L-lysine (L-Lys) to L-PA. This suggested that there is a gene encoding ""P6C reductase"" that catalyzes the reduction of P6C to L-PA in the genome of E. coli. The complementation experiment of proC32 in E. coli RK4904 for L-PA production clearly shows that the expression of both lat and proC is essential for the biotransformation of L-Lys to L-PA. Further, We showed that both LAT and pyrroline-5-carboxylate (P5C) reductase, the product of proC, were needed to convert L-Lys to L-PA in vitro. These results demonstrate that P5C reductase catalyzes the reduction of P6C to L-PA. Biotransformation of L-Lys to L-PA using lat-expressing E. coli BL21 was done and L-PA was accumulated in the medium to reach at an amount of 3.9g/l after 159 h of cultivation. It is noteworthy that the ee-value of the produced pipecolic acid was 100%.