Efficient Nepsilon-lauroyl-L-lysine production by recombinant epsilon-lysine acylase from Streptomyces mobaraensis.
J Biotechnol
; 141(3-4): 160-5, 2009 May 20.
Article
en En
| MEDLINE
| ID: mdl-19433221
epsilon-Lysine acylase from Streptomyces mobaraensis (Sm-ELA), which specifically catalyzes hydrolysis of the epsilon-amide bond in various Nepsilon-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 55,816Da. 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 2500-2800U/mg, which were similar to that obtained for the wild-type Sm-ELA. Using the cell-free extract of the recombinant S. lividans cells, Nepsilon-lauroyl-L-lysine was synthesized from 500mM L-lysine hydrochloride and 50, 100, or 250mM lauric acid in an aqueous buffer solution at 37 degrees C. The yields were close to 100% after 6 and 9h of reaction for 50 and 100mM lauric acid, respectively, and 90% after 24h for 250mM lauric acid.
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1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Streptomyces
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Amidohidrolasas
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Lisina
Idioma:
En
Revista:
J Biotechnol
Asunto de la revista:
BIOTECNOLOGIA
Año:
2009
Tipo del documento:
Article
País de afiliación:
Japón
Pais de publicación:
Países Bajos