Crystal structures of K33 mutant hen lysozymes with enhanced activities.
作者： T. Goto , T. Ohkuri , S. Shioi , Y. Abe , T. Imoto
DOI: 10.1093/JB/MVN108
关键词: Lysozyme 、 Biology 、 Mutagenesis (molecular biology technique) 、 Mutant 、 Biochemistry 、 Mutation testing 、 Chitin 、 Micrococcus luteus 、 Side chain 、 Enzyme assay
摘要: Using random mutagenesis, we previously obtained K33N mutant lysozyme that showed a large lytic halo on the plate coating Micrococcus luteus. In order to examine effects of mutation enzyme activity, prepared and K33A lysozymes from yeast. It was found activities both were higher than those wild-type based results activity measurements against M. luteus (lytic activity) glycol chitin. Moreover, 3D structures solved by X-ray crystallographic analyses. The side chain K33 in hydrogen bonded with N37 involved substrate-binding region, orientation different lysozyme. These suggest enhancement due an alteration N37. On other hand, less stable Lysozyme may sacrifice its acquire conformational stability at position 33.
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DianeM Retallack, TraceyC Thomas, Ying Shao, KeithL Haney, SolM Resnick, VincentD Lee, CharlesH Squires, Identification of anthranilate and benzoate metabolic operons of Pseudomonas fluorescens and functional characterization of their promoter regions Microbial Cell Factories. ,vol. 5, pp. 1- 13 ,(2006) , 10.1186/1475-2859-5-1
PratulK Agarwal, Enzymes: An integrated view of structure, dynamics and function Microbial Cell Factories. ,vol. 5, pp. 2- 2 ,(2006) , 10.1186/1475-2859-5-2
J.F. Krebs, C.A. Fierke, Determinants of catalytic activity and stability of carbonic anhydrase II as revealed by random mutagenesis. Journal of Biological Chemistry. ,vol. 268, pp. 948- 954 ,(1993) , 10.1016/S0021-9258(18)54025-4
S Kanaya, M Itaya, Expression, purification, and characterization of a recombinant ribonuclease H from Thermus thermophilus HB8. Journal of Biological Chemistry. ,vol. 267, pp. 10184- 10192 ,(1992) , 10.1016/S0021-9258(19)50217-4
I Kumagai, F Sunada, S Takeda, K Miura, Redesign of the Substrate-binding Site of Hen Egg White Lysozyme Based on the Molecular Evolution of C-type Lysozymes* Journal of Biological Chemistry. ,vol. 267, pp. 4608- 4612 ,(1992) , 10.1016/S0021-9258(18)42876-1
K Yutani, K Ogasahara, T Tsujita, K Kanemoto, M Matsumoto, S Tanaka, T Miyashita, A Matsushiro, Y Sugino, E W Miles, Tryptophan synthase alpha subunit glutamic acid 49 is essential for activity. Studies with 19 mutants at position 49. Journal of Biological Chemistry. ,vol. 262, pp. 13429- 13433 ,(1987) , 10.1016/S0021-9258(19)76444-8
R Kuroki, H Yamada, T Moriyama, T Imoto, Chemical mutations of the catalytic carboxyl groups in lysozyme to the corresponding amides. Journal of Biological Chemistry. ,vol. 261, pp. 13571- 13574 ,(1986) , 10.1016/S0021-9258(18)67057-7
Yoshio Hashimoto, Takeyoshi Miki, Masami Mukae, Tadashi Ueda, Taiji Imoto, Construction of a yeast expression system with positive selection for gene insertion in the absence of a specific phenotype Gene. ,vol. 207, pp. 167- 170 ,(1998) , 10.1016/S0378-1119(97)00621-5
I. Barry Vipond, Stephen E. Halford, Random Mutagenesis Targeted to the Active Site of the EcoRV Restriction Endonuclease Biochemistry. ,vol. 35, pp. 1701- 1711 ,(1996) , 10.1021/BI952391D
Axel T Brünger, Paul D Adams, G Marius Clore, Warren L DeLano, Piet Gros, Ralf W Grosse-Kunstleve, J-S Jiang, John Kuszewski, Michael Nilges, Navraj S Pannu, Randy J Read, Luke M Rice, Thomas Simonson, Gregory L Warren, Crystallography & NMR System: A New Software Suite for Macromolecular Structure Determination Acta Crystallographica Section D-biological Crystallography. ,vol. 54, pp. 905- 921 ,(1998) , 10.1107/S0907444998003254