Skip Navigation



Journal of Biochemistry Advance Access published online on July 23, 2007
Journal of Biochemistry, doi:10.1093/jb/mvm134
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
142/3/317    most recent
mvm134v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Inouye, K.
Right arrow Articles by Yasukawa, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Inouye, K.
Right arrow Articles by Yasukawa, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2007 The Japanese Biochemical Society

Effects of Neutral Salts and Alcohols on the Activity of Streptomyces caespitosus Neutral Protease (ScNP)

Kuniyo Inouye*, Takashi Shimada and Kiyoshi Yasukawa

Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502

*To whom correspondence should be addressed: Tel: +81-75-753-6266, Fax: +81-75-753-6265, E-mail: inouye{at}kais.kyoto-u.ac.jp

Received May 9, 2007; Accepted June 12, 2007


  Abstract

Streptomyces caespitosus neutral protease (ScNP) is one of the smallest metalloproteinase with a molecular mass of 14 kDa. Effects of solvent composition on ScNP activity were examined using a peptide substrate. The kcat/Km values of ScNP exhibited bell-shaped pH-dependence with the optimal pH of 6.4-7.0 and the pKa values of 5.0 ± 0.1 and 8.3 ± 0.1. ScNP activity increased in an exponential fashion with increasing [NaCl]. The relative kcat/Km value at 3.6 M NaCl to that at 0 M NaCl was 3.7, and the degree of the activation at x M NaCl was expressed as 1.2 x (x < 2.0) and 1.4x (x > 2.0). On the other hand, ScNP activity decreased with increasing concentrations of LiCl, KCl, NaBr, LiBr, KBr, and NaClO4. Alcohols inhibited ScNP activity with the IC50 values, the concentration required for decreasing the activity at 50% of the maximum, of 0.77-6.54 M. The order of the inhibitory potency was 1-butanol, 2-methyl-1-propanol, 2-methyl-2-butanol > 2-methyl-2-propanol, 2-butanol, 1-propanol > 2-propanol ethanol methanol. The activities recovered completely by the dilution of alcohols, suggesting that the ScNP inhibition by alcohols is reversible. These characteristics of ScNP are compared with those of human matrix metalloproteinase 7 and thermolysin.

Key Words: alcohol, metalloproteinase, salt effect, pH-activity profile, Streptomyces caespitosus neutral protease


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J BiochemHome page
M. Kusano, K. Yasukawa, and K. Inouye
Insights into the Catalytic Roles of the Polypeptide Regions in the Active Site of Thermolysin and Generation of the Thermolysin Variants with High Activity and Stability
J. Biochem., January 1, 2009; 145(1): 103 - 113.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.