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Structure-Functionality of Egg-White Lysozyme by Protein Engineering

Research Project

Project/Area Number 02660143
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 製造化学・食品
Research InstitutionYamaguchi University

Principal Investigator

KATO Akio  Yamaguchi Univ., Fac. of Agriculture, Professor, 農学部, 教授 (00035114)

Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsProtein Engineering / Lysozyme / Deamidation / Emulsifying Properties / Foaming Properties / 変異型リゾチ-ム
Research Abstract

To investigate the structural and functional properties of lysozyme, various mutant enzymes were constructed by protein engineering. Lysozyme mutants deamidated at positions 103 (NlO3D) and 106 (NlO6D), cleaved salt linkage between Iys 13 and leu 129 (K13D), and deleted SS bond between positions 96 and 74 (C96/74A) were prepared. The wild-type and mutant lysozymes were expressed in Saccharomyces cerevisiae and purified from the cultivation medium in two steps by cation exchange chromatography on CM-Toyopearl. The lytic activities of each mutant lysozymes were almost the same as that of wild lysozyme, although the optimal pH of activity was slightly shifted to lower pH by the deamidation. -The Gibbs free energy changes of unfolding(DELTAG)at 20゚C for N103D and N106D were almost the same as that of wild-type. On the other hand, the structural flexibility of lysozymes, estimated by protease digestion, was significantly increased by the deamidation. The surface functional properties of deamidated lysozymes were considerably enhanced, compared to those of wild-type lysozyme. These results suggest that structural flexibility is an important governing factor in surface functional properties of proteins, regardless of their structural stability. On the other hand, DELTA G of mutant K13D and C96/74A was much lower than that of wild-type lysozyme, suggesting their unstable structure. These mutants had better surface properties than deamidated lysozymes. Thus, protein stability seems to be more effective factor than its flexibility for the surface functional properlies of proteins.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] A.Kato,S.Tanimoto,Y.Muraki,K.Kobayashi,I.Kumagai: "Structural and functional properties of hen egg-white lysozyme deamidated by protein engineering." Biosci.Biotech.Biochem.56. (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S.Nakamura,A.Kato,K.Kobayashi: "New antimicrobial characteristics of lysozyme-dextran conjugate." J.Agric.Food Chem.39. 647-650 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hisham R.Ibrahim,A.Kato,K.Kobayashi: "Antimicrobial effects of lysozyme against Gram-negative bacteria due to covalent binding of palmitic acid." J.Agric.Food Chem.39. 2077-2082 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] A. Kato, S. Tanimoto, Y. Muraki, K. Kobayashi and I. Kumagai.: "Structural and Functional Properties of Hen Egg-White Lysozyme Deamidated by Protein Engineering" Biosci. Biotech. Biochem. 56. (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Nakamura, A. Kato and K. Kobayashi: "New Antimicrobial Characteristics of Lysozyme-Dextran Conjugate." J. Agric. Food Chem.39. 647-650 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H. R. Ibrahim, A. Kato and K. Kobayashi: "Antimicrobial Effects of Lysozyme against Gram-Negative Bacteria Due to Covalent Binding of Parmitic Acid." J. Agric. Food Chem.39. 2077-2082 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Akio Kato,Kunihiko Kobayashi,Yoshifumi Muraki,Shoto Tanimoto,Izumi Kumagai: "Structural and Functional Properties of Hen Eggーwhite Lysozyme Deamidated by Protein Engineering." Biosci.Biotech.Biochem.56. (1992)

    • Related Report
      1991 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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