Immobilization of N-Carbamyl-D-Amino Acid Amidohydrolase

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概要

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• N-Carbamyl-D-amino acid amidohydrolase (DCase), produced with recombinant Escherichia coli cells using a cloned gene from Agrobacterium sp. strain KNK712,has been immobilized for use in the production of D-amino acids. The porous polymers, Duolite A-568 and Chitopearl 3003,were much better than other resins for the activity and stability of the adsorbed enzyme. The activity of DCase expressed on Duolite A-568 and Chitopearl 3003 amounted to 96 units/g-wet-resin and 91 units/g-wet-resin, respectively. DCase immobilized on Duolite A-568 was found to be most stable at about pH 7,and it was further stabilized by reductants such as dithiothreitol, L-cysteine, cysteamine, and sodium hydrosulfite. The stability during the repeated batch reactions was greatly improved when dithiothreitol was in the reaction mixture, and the higher crosslinking degree with glutaraldehyde also stabilized the immobilized enzyme. After 14 times repeated reactions, the remaining activity of the immobilized enzyme cross-linked with 0.1% and 0.2% of glutaraldehyde, and 0.2% of glutaraldehyde with dithiothreitol in the reaction mixture was 12%, 18%, and 63%, respectively. DCase produced with Pseudomonas sp. strain KNK003A and Pseudomonas sp. strain KNK505,which are thermotolerant soil bacteria, and that with Agrobacterium sp. strain KNK712 were also immobilized on Duolite A-568. The stability of the enzymes of thermotolerant bacteria during reactions was superior to that of Agrobacterium sp. strain KNK712,though the activity was lower than that of strain KNK712.

• 社団法人日本農芸化学会の論文
• 1998-10-23

著者

• Ikenaka Y

  Fine Chemicals Research Laboratories Kaneka Corporation

• Yamada Y

  Doshisha Women's Coll. Liberal Arts Kyoto Jpn

• Yamada Y

  The Department Of Mechanical Engineering And Intelligent Systems University Of Electro-communication

• NANBA Hirokazu

  Fine Chemicals Research Laboratories, Kaneka Corporation

• Nanba H

  Kaneka Corp. Hyogo Jpn

• Nanba Hirokazu

  Fine Chemicals Research Laboratories Kaneka Corporation

• Yamashita Kenji

  Biochemical Research Laboratories Kanegafuchi Chemical Industry Co. Ltd.

• Takahashi S

  Akita Res. Inst. Food And Brewing Akita

• Ikenaka Yasuhiro

  Biochemical Research Laboratories Kanegafuchi Chemical Industry Co. Ltd.

• Yamada Yukio

  Fine Chemicals Research Laboratories Kaneka Corporation

• IKENAKA YASUHIRO

  Fine Chemicals Research Laboratories, Kaneka Corporation

• YAJIMA KAZUYOSHI

  Fine Chemicals Research Laboratories, Kaneka Corporation

• TAKANO MASAYUKI

  Fine Chemicals Research Laboratories, Kaneka Corporation

• TAKAHASHI SATOMI

  Fine Chemicals Research Laboratories, Kaneka Corporation

• OHKUBO Kazuma

  Fine Chemicals Research Laboratories, Kaneka Corporation

• HIRAISHI Yoshirou

  Fine Chemicals Research Laboratories, Kaneka Corporation

• YAMADA Kazuhiko

  Fine Chemicals Research Laboratories, Kaneka Corporation

• Ohkubo Kazuma

  Fine Chemicals Research Laboratories Kaneka Corporation

• Takano Masayuki

  Fine Chemicals Research Laboratories Kaneka Corporation

• Yamada Yasuyuki

  Graduate School Of Biological Science Nara Institute Of Science And Technoiogy

• Yajima Kazuyoshi

  Fine Chemicals Research Laboratories Kaneka Corporation

• Ikenaka Yasuhiro

  Fine Chemicals Research Laboratories Kaneka Corporation

• Hiraishi Yoshirou

  Fine Chemicals Research Laboratories Kaneka Corporation

• Yamada Yukio

  Mechanical Engineering Laboratory Material Science And Bioengineering Department Aist

• Yamada K

  Faculty Of Engineering Okayama University:(present Address)department Of Biotechnology And Life Scie

• Yamada Y

  Fac. Of Education Utsunomiya Univ.

• Yamada Y

  Faculty Of Education Utsunomiya University

• Katayama Y

  Graduate School Of Bio-applications And Systems Engineering Tokyo University Of Agriculture And Tech

• Suzuki Takahito

  Biological Laboratory Faculty Of Science Nara Women's University

• Yamada Yuko

  Kyoto Univ. Kyoto Jpn

• Yamaguchi Yuko

  Laboratory For Molecular Functions Of Food Division Of Food Science And Biotechnology Graduate Schoo

• Nanba Hirokazu

  Fine Chemicals Research Laboratories

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