Crystal structures of K33 mutant hen lysozymes with enhanced activities

Takashi Goto, Takatoshi Ohkuri, Seijiro Shioi, Yoshito Abe, Taiji Imoto, Tadashi Ueda

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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 the effects of mutation of K33N on enzyme activity, we prepared K33N and K33A mutant lysozymes from yeast. It was found that the activities of both the mutant lysozymes were higher than those of the wild-type lysozyme based on the results of the activity measurements against M. luteus (lytic activity) and glycol chitin. Moreover, 3D structures of K33N and K33A mutant lysozyme were solved by X-ray crystallographic analyses. The side chain of K33 in the wild-type lysozyme hydrogen bonded with N37 involved in the substrate-binding region, and the orientation of the side chain of N37 in K33 mutant lysozymes were different in the wild-type lysozyme. These results suggest that the enhancement of activity in K33N mutant lysozyme was due to an alteration in the orientation of the side chain of N37. On the other hand, K33N lysozyme was less stable than the wild-type lysozyme. Lysozyme may sacrifice its enzyme activity to acquire the conformational stability at position 33.

Original languageEnglish
Pages (from-to)619-623
Number of pages5
JournalJournal of biochemistry
Volume144
Issue number5
DOIs
Publication statusPublished - Nov 1 2008

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Muramidase
Crystal structure
Micrococcus luteus
Enzyme activity
Mutagenesis
Enzymes
Yeast
Hydrogen
Yeasts
X-Rays
X rays
Coatings

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Crystal structures of K33 mutant hen lysozymes with enhanced activities. / Goto, Takashi; Ohkuri, Takatoshi; Shioi, Seijiro; Abe, Yoshito; Imoto, Taiji; Ueda, Tadashi.

In: Journal of biochemistry, Vol. 144, No. 5, 01.11.2008, p. 619-623.

Research output: Contribution to journalArticle

Goto, Takashi ; Ohkuri, Takatoshi ; Shioi, Seijiro ; Abe, Yoshito ; Imoto, Taiji ; Ueda, Tadashi. / Crystal structures of K33 mutant hen lysozymes with enhanced activities. In: Journal of biochemistry. 2008 ; Vol. 144, No. 5. pp. 619-623.
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