Efficient Nε-lauroyl-l-lysine production by recombinant ε-lysine acylase from Streptomyces mobaraensis

Mayuko Koreishi, Ryoko Kawasaki, Hiroyuki Imanaka, Koreyoshi Imamura, Yasuaki Takakura, Kazuhiro Nakanishi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

ε-Lysine acylase from Streptomyces mobaraensis (Sm-ELA), which specifically catalyzes hydrolysis of the ε-amide bond in various Nε-acyl-l-lysines, was cloned and sequenced. The Sm-ELA gene consists of a 1617-bp open reading frame that encodes a 538-amino acid protein with a molecular mass of 55,816 Da. An NCBI protein-protein BLAST search revealed that the enzyme belongs to the YtcJ-like metal-dependent amidohydrolase family, which is further characterized as the metallo-dependent hydrolase superfamily. The Sm-ELA gene was ligated into a pUC702 vector for expression in Streptomyces lividans TK24. Expression of recombinant Sm-ELA in S. lividans was approximately 300-fold higher than that in wild-type S. mobaraensis. The recombinant Sm-ELAs from the cell-free extract and culture supernatant were purified to homogeneity. The specific activities of the purified Sm-ELAs were 2500-2800 U/mg, which were similar to that obtained for the wild-type Sm-ELA. Using the cell-free extract of the recombinant S. lividans cells, Nε-lauroyl-l-lysine was synthesized from 500 mM l-lysine hydrochloride and 50, 100, or 250 mM lauric acid in an aqueous buffer solution at 37 °C. The yields were close to 100% after 6 and 9 h of reaction for 50 and 100 mM lauric acid, respectively, and 90% after 24 h for 250 mM lauric acid.

Original languageEnglish
Pages (from-to)160-165
Number of pages6
JournalJournal of Biotechnology
Volume141
Issue number3-4
DOIs
Publication statusPublished - May 20 2009

Fingerprint

amidase
lauric acid
Streptomyces
Streptomyces lividans
Lysine
Proteins
Acids
Genes
Cell Extracts
Hydrolases
Amidohydrolases
Molecular mass
Amides
Amino acids
Hydrolysis
Enzymes
Open Reading Frames
Buffers
Cell Culture Techniques
Metals

Keywords

  • ε-Lysine acylase
  • Actinomycete
  • Nε-Lauroyl-l-lysine
  • Streptomyces mobaraensis

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Efficient Nε-lauroyl-l-lysine production by recombinant ε-lysine acylase from Streptomyces mobaraensis. / Koreishi, Mayuko; Kawasaki, Ryoko; Imanaka, Hiroyuki; Imamura, Koreyoshi; Takakura, Yasuaki; Nakanishi, Kazuhiro.

In: Journal of Biotechnology, Vol. 141, No. 3-4, 20.05.2009, p. 160-165.

Research output: Contribution to journalArticle

Koreishi, Mayuko ; Kawasaki, Ryoko ; Imanaka, Hiroyuki ; Imamura, Koreyoshi ; Takakura, Yasuaki ; Nakanishi, Kazuhiro. / Efficient Nε-lauroyl-l-lysine production by recombinant ε-lysine acylase from Streptomyces mobaraensis. In: Journal of Biotechnology. 2009 ; Vol. 141, No. 3-4. pp. 160-165.
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