Cloning and expression of L-N-carbamoylase gene from Arthrobacter BT801 in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
; (12): 174-177, 2003.
Article
en Zh
| WPRIM
| ID: wpr-270118
Biblioteca responsable:
WPRO
ABSTRACT
Hydantoin-utility-enzyme is widely used in enzymic production of various amino acids. One of its component, carbamoylase, is responsible for the conversion of N-carbamylamino acids to corresponding amino acids, which is crucial for the stereoselectivity and rate limiting. To improve the production of the enzyme, an L-N-carbamoylase gene from Arthrobacter BT801, a hydantoinase producting strain being able to convert 5-benzylhydantoin to phenylalanine, was cloned into E. coli. The gene was highly expressed in E. coli M15 under control of T5 promoter. A protein band about 44kD was detected by SDS-PAGE in the recombinant cell lysate. The objective product, which is principally in soluble form, represented 40% of total cell protein. The N-carbamoylase specific activity of the recombinant M15/pQE60- hyuC is 53 times higher than that of Arthrobacter BT801. The total biotransformation activity increased 8.1 times when. M15/pQE60-hyuC was added into the Arthrobacter BT801 reaction system. The successful expression of the enzyme is significant for the application of the hydantoinase producing strain or the enzyme thereof.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Fenilalanina
/
Plásmidos
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Arthrobacter
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Reacción en Cadena de la Polimerasa
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Electroforesis en Gel de Poliacrilamida
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Escherichia coli
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Amidohidrolasas
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Vectores Genéticos
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Genética
/
Hidantoínas
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2003
Tipo del documento:
Article