Role of arginine residues of D-aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6

Kazuaki Yoshimune, Masanori Kanda, Mamoru Wakayama, Shun Ichi Kanda, Akiko Sato, Kenji Sakai, Mitsuaki Moriguchi

研究成果: Contribution to journalArticle査読

抄録

To investigate the role of arginine in the folding of D-aminoacylase, seven arginine residues, R26, R152, R296, R302, R354, R377, and R391, among twelve arginine residues highly conserved in D-aminoacylase, N-acyl-D-aspartatc amidohydrolase (D-AAase), and N-acyl-D-glutamate amidohydrolase (D-AGase) from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (Alcaligenes A-6) were substituted with lysine by site-directed mutagenesis. The mutants, R26K, R152K, R296K, and R302K were identified as mutations that increase partitioning of the enzyme into inclusion bodies. No mutants with substitutions within the carboxyterminal segment were found to increase partitioning into inclusion bodies (R354K, R377K, and R392K). These results suggest that arginine residues that position between the N-terminus and central region can play an important role in facilitating folding or stabilizing the structure of D-aminoacylase. By anaerobic cultivation, the production level of R302K in the soluble fraction was improved. Coexpression of the DnaK-DnaJ-GrpE chaperone assisted the folding of R302K, and reduced the effect of the aeration conditions on the solubility of R302K. We hypothesized that R302K requires a larger amount of chaperones for efficient folding than the wild type enzyme.

本文言語英語
ページ(範囲)289-294
ページ数6
ジャーナルProtein and Peptide Letters
12
3
DOI
出版ステータス出版済み - 4 1 2005
外部発表はい

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biochemistry

フィンガープリント 「Role of arginine residues of D-aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

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