Document Detail

Stabilization of invertase by molecular engineering.
MedLine Citation:
PMID:  19918887     Owner:  NLM     Status:  MEDLINE    
Extracellular invertase (EC of Saccharomyces cerevisiae was stabilized against thermal denaturation by intermolecular and intramolecular crosslinking of the surface nucleophilic functional groups with diisocyanate homobifunctional reagents (O==C==N(CH(2))(n)N==C==O) of various lengths (n = 4, 6, 8). Crosslinking with 1,4-diisocyanatobutane (n = 4) proved most effective in enhancing thermostability. Stability was improved dramatically by crosslinking 0.5 mg/mL of protein with 30 mumol/mL of the reagent. Molecular engineering by crosslinking reduced the first-order thermal denaturation constant at 60 degrees C from 1.567 min(-1) (for the native enzyme) to 0.437 min(-1) (for the stabilized enzyme). Similarly, the best crosslinking treatment increased the activation energy for denaturation from 391 kJ mol(-1) (for the native protein) to 466 kJ mol(-1) (for the stabilized enzyme). Crosslinking was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis.
Pattamawadee Tananchai; Yusuf Chisti
Related Documents :
2105527 - Characterization of murine liver-derived inhibitory protein.
2793827 - Enzymatic dehalogenation of pentachlorophenol by extracts from arthrobacter sp. strain ...
10746197 - Influence of incubation temperature on activity of ligninolytic enzymes in sterile soil...
21994937 - A new family of carbohydrate esterases is represented by a gdsl hydrolase/acetylxylan e...
21616177 - Dual diaminopimelate biosynthesis pathways in bacteroides fragilis and clostridium ther...
18023917 - Phenylalanine ammonia-lyase-mediated biosynthesis of 2-hydroxy-4-methoxybenzaldehyde in...
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology progress     Volume:  26     ISSN:  1520-6033     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2010 Jan-Feb
Date Detail:
Created Date:  2010-02-15     Completed Date:  2010-05-24     Revised Date:  2011-05-26    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  111-7     Citation Subset:  IM    
School of Engineering, Massey University, Palmerston North, New Zealand.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Chemical Engineering*
Cross-Linking Reagents / chemistry*,  pharmacology
Electrophoresis, Polyacrylamide Gel
Enzyme Stability / drug effects
Hot Temperature
Protein Denaturation / drug effects
Saccharomyces cerevisiae / enzymology
beta-Fructofuranosidase / chemistry*,  metabolism*
Reg. No./Substance:
0/Cross-Linking Reagents; EC

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  An MR-compatible bicycle ergometer for in-magnet whole-body human exercise testing.
Next Document:  Change in the proton T(1) of fat and water in mixture.