Characterization of an [alpha]-amino--caprolactam racemase with broad substrate specificity from Citreicella sp. SE45

• Published in: Journal of industrial microbiology & biotechnology Vol. 44; no. 4-5; p. 677
• Main Authors: Payoungkiattikun, Wisarut, Okazaki, Seiji, Ina, Atsutoshi, H-kittikun, Aran, Asano, Yasuhisa
• Format: Journal Article
• Language: English
• Published: Fairfax Oxford University Press 01.05.2017
• Subjects: Amides; Amino acids; Biotechnology; Cloning; E coli; Enzymes; Plasmids; Proteins; Sodium
• ISSN: 1367-5435; 1476-5535
• DOI: 10.1007/s10295-016-1825-8

[alpha]-Amino-[straight epsilon]-caprolactam (ACL) racemizing activity was detected in a putative dialkylglycine decarboxylase (EC 4.1.1.64) from Citreicella sp. SE45. The encoding gene of the enzyme was cloned and transformed in Escherichia coli BL21 (DE3). The molecular mass of the enzyme was shown to be 47.4 kDa on SDS-polyacrylamide gel electrophoresis. The enzymatic properties including pH and thermal optimum and stabilities were determined. This enzyme acted on a broad range of amino acid amides, particularly unbranched amino acid amides including l-alanine amide and l-serine amide with a specific activity of 17.5 and 21.6 U/mg, respectively. The K m and V max values for d- and l-ACL were 5.3 and 2.17 mM, and 769 and 558 [mu]mol/min.mg protein, respectively. Moreover, the turn over number (K cat) and catalytic efficiency (K cat/K m) of purified ACL racemase from Citreicella sp. SE45 using l-ACL as a substrate were 465 S-1 and 214 S-1mM-1, respectively. The new ACL racemase from Citreicella sp. SE45 has a potential to be used as the biocatalytic application.