Role of Conserved Histidine Residues in D-Aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6.
-
- WAKAYAMA Mamoru
- Department of Applied Chemistry, Faculty of Engineering
-
- YADA Harutaka
- Department of Applied Chemistry, Faculty of Engineering
-
- KANDA Shun-ichi
- Department of Applied Chemistry, Faculty of Engineering
-
- HAYASHI Shin-ichi
- Department of Applied Chemistry, Faculty of Engineering
-
- YATSUDA Yukinori
- Department of Applied Chemistry, Faculty of Engineering
-
- SAKAI Kenji
- Department of Applied Chemistry, Faculty of Engineering
-
- MORIGUCHI Mitsuaki
- Department of Applied Chemistry, Faculty of Engineering
Bibliographic Information
- Other Title
-
- Role of Conserved Histidine Residues in<scp>D</scp>-Aminoacylase from<i>Alcaligenes xylosoxydans</i>subsp.<i>xylosoxydans</i>A-6
Search this article
Abstract
D-Aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (Alcaligenes A-6) was strongly inactivated by diethylpyrocarbonate (DEPC). An H67N mutant was barely active, with a kcat/Km 6.3×104 times lower than that of the recombinant wild-type enzyme, while the H67I mutant lost detectable activity. The H67N mutant had almost constant Km, but greatly decreased kcat. These results suggested that His67 is essential to the catalytic event. Both H69N and H69I mutants were overproduced in the insoluble fraction. The kcat/Km of H250N mutant was reduced by a factor of 2.5×104-fold as compared with the wild-type enzyme. No significant difference between H251N mutant and wild-type enzymes in the Km and kcat was found. The Zn content of H250N mutant was nearly half of that of wild-type enzyme. These results suggest that the His250 residue might be essential to catalysis via Zn binding.<br>
Journal
-
- Bioscience, Biotechnology, and Biochemistry
-
Bioscience, Biotechnology, and Biochemistry 64 (1), 1-8, 2000
Japan Society for Bioscience, Biotechnology, and Agrochemistry
- Tweet
Details 詳細情報について
-
- CRID
- 1390282681450751616
-
- NII Article ID
- 110002679739
-
- NII Book ID
- AA10824164
-
- DOI
- 10.1271/bbb.64.1
-
- COI
- 1:CAS:528:DC%2BD3cXhtVyitL0%3D
-
- ISSN
- 13476947
- 09168451
-
- NDL BIB ID
- 5142906
-
- PubMed
- 10705441
-
- Text Lang
- en
-
- Data Source
-
- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
-
- Abstract License Flag
- Disallowed