Biotransformation ofL-Lysine toL-Pipecolic Acid Catalyzed byL-Lysine 6-Aminotransferase and Pyrroline-5-carboxylate Reductase

Molecular characterization of lysine 6-dehydrogenase from Achromobacter denitrificans

“…The crystallization of the A. denitrificans enzyme will help in the establishment of the three-dimensional structure of Lys 6-DH. Recently, much attention has been paid to the asymmetric synthesis of L-pipecolate (18,19), a key component of the immunosuppressant rapamycin (20) and the antitumor agent sandramycin (21). L-Pipecolate can be derived from P-6-C (18).…”

Enzymatic Synthesis ofL-Pipecolic Acid by Δ1-Piperideine-2-carboxylate Reductase fromPseudomonas putida

“…An enzymatic system for the synthesis of L-pipecolic acid from L-lysine by commercial L-lysine -oxidase from Trichoderma viride and an extract of recombinant Escherichia coli cells coexpressing Á 1 -piperideine-2-carboxylate reductase from Pseudomonas putida and glucose dehydrogenase from Bacillus subtilis is described. A laboratory-scale process provided 27 g/l of Lpipecolic acid in 99.7% e.e.Key words: Á 1 -piperideine-2-carboxylate reductase; Pseudomonas putida; L-pipecolic acid; Á 1 -piperideine-2-carboxylate L-Pipecolic acid, a nonproteinogenic -amino acid, is a key component of many bioactive molecules, such as the immunosuppressant FK506, 1) the anticancer agent VX710, 2) the antifungal antibiotic demethoxyrapamycin,3) the N-methyl-D-aspartate antagonist selfotel, 4) the antitumor antibiotic sandramycin, 5) the phytotoxic metabolite Cyl-2, 6) the anesthetic bupivacaine, 7) and the HIV protease inhibitor palinavir.8) There is an increasing demand for a convenient and efficient synthetic route to enantiomerically pure L-pipecolic acid because of its use in the synthesis of new medicaments.Current methods to obtain a pure enantiomer of L-pipecolic acid involve chemical resolution, 9) stereoselective transformation, 10) the derivatization of natural amino acids, 11) and enzymatic reactions, [12][13][14][15][16] but most of these methods fail to provide a satisfactory solution for the synthesis of chiral pipecolic acid on an industrial scale due to certain limitations, such as tedious procedures, low yields, and unavailability of starting materials. Hence, new and convenient methods for the preparation of optically active pipecolic acid are required.…”

“…Current methods to obtain a pure enantiomer of L-pipecolic acid involve chemical resolution, 9) stereoselective transformation, 10) the derivatization of natural amino acids, 11) and enzymatic reactions, [12][13][14][15][16] but most of these methods fail to provide a satisfactory solution for the synthesis of chiral pipecolic acid on an industrial scale due to certain limitations, such as tedious procedures, low yields, and unavailability of starting materials. Hence, new and convenient methods for the preparation of optically active pipecolic acid are required.…”

Increase in the Rate ofL-Pipecolic Acid Production Usinglat-ExpressingEscherichia colibylysPandyeiEAmplification

“…1) In a further study, we tried to construct microorganisms with a higher activity for L-PA production. To improve the L-PA production rate, weˆrst increased the expression level of lat or proC by amplifying the copy number of each gene.…”

“…A fermentation experiment was done using these strains in the manner described previously. 1) One hundred microliters of each culture was collected after 15, 39, 63, 87, 111, and 159 h of cultivation to measure the amount of L-PA by HPLC as described previously. 1) At each sampling time, 500 ml of 50z glycerol and 500 ml of 50z L-lysine HCl were fed in the culture, and the cultivation was continued.…”