Journal
JOURNAL OF BIOTECHNOLOGY
Volume 129, Issue 4, Pages 645-650Publisher
ELSEVIER
DOI: 10.1016/j.jbiotec.2007.02.001
Keywords
nitrilase; mandelonitrile; rational genome mining; Bradyrhizobium japonicum
Categories
Ask authors/readers for more resources
A mandelonitrile hydrolase bll6402 from Bradyrhizobium japonicum USDA 110 was predicted by rational genome mining, i.e. combining traditional genome mining with functional analysis of the genetic organization of the putative nitrilase gene within the chromosome of microorganisms. This putative gene was cloned and over-expressed in Escherichia coli, and the encoded protein was purified to give a nitrilase with a molecular mass of about 37 kDa. The molecular weight of the holoenzyme was about 455 kDa, suggesting that nitrilase bll6402 self-aggregated to the active form with native structure being 12 subunits of identical size. This nitrilase was most active toward mandelonitrile with V-max and K-m for mandelonitrile being 44.7 U/mg and 0.26 mM, respectively. The k(cat) and overall catalytic efficiency k(cat)/K-m were 27.0 s(-1) and 1.04 x 10(5) M-1 s(-1), indicating that nitrilase bll6402 is very active for the hydrolysis of mandelonitrile to mandelic acid. Nitrilase b116402 also effectively hydrolyzed several mandelonitrile derivatives. (c) 2007 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available