Reengineering of subtilisin Carlsberg for oxidative resistance
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Ljubica Vojcic
Abstract
Mild bleaching conditions by in situ production of hydrogen peroxide or peroxycarboxylic acid is attractive for pulp, textile, and cosmetics industries. The enzymatic generation of chemical oxidants is often limited by enzyme stability. The subtilisin Carlsberg variant T58A/L216W/M221 is a promiscuous protease that was improved in producing peroxycarboxylic acids. In the current article, we identified two amino acid positions (Trp216 and Met221) that are important for the oxidative resistance of subtilisin Carlsberg T58A/L216W/M221. Site-saturation mutagenesis at positions Trp216 and Met221, which are located close to the active site, resulted in variants M4 (T58/W216M/M221) and M6 (T58A/W216L/M221C). Variants M4 (T58/W216M/M221) and M6 (T58A/W216L/M221C) have a 2.6-fold (M4) and 1.5-fold (M6) increased oxidative resistance and 1.4-fold increased kcat values for peroxycarboxylic acid formation, compared with wild-type subtilisin Carlsberg.
©2012 by Walter de Gruyter Berlin Boston
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Artikel in diesem Heft
- Masthead
- Masthead
- Reviews
- Exosomes: the ideal nanovectors for biodelivery
- Biological applications of the electrochemical sensing of nitric oxide: fundamentals and recent developments
- Minireview
- Cerebral cavernous malformation is a vascular disease associated with activated RhoA signaling
- Research Articles/Short Communications
- Protein Structure and Function
- Diversity, abundance, and sex-specific expression of chemosensory proteins in the reproductive organs of the locust Locusta migratoria manilensis
- Structure of the Ca2+-saturated C-terminal domain of scallop troponin C in complex with a troponin I fragment
- An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
- Reengineering of subtilisin Carlsberg for oxidative resistance
- New insight into the molecular switch mechanism of human Rho family proteins: shifting a paradigm
- Cell Biology and Signaling
- HbG200-mediated preinduction of heme oxygenase-1 improves bile flow and ameliorates pericentral downregulation of Bsep and Mrp2 following experimental liver ischemia and reperfusion
- Effect of silver nanoparticles on human primary keratinocytes
- Multifunctional silica nanoparticles for optical and magnetic resonance imaging
- Proteolysis
- Activation profiles of human kallikrein-related peptidases by matrix metalloproteinases
