Document Detail


Stabilization of invertase by molecular engineering.
MedLine Citation:
PMID:  19918887     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Extracellular invertase (EC 3.2.1.26) 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.
Authors:
Pattamawadee Tananchai; Yusuf Chisti
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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    
Affiliation:
School of Engineering, Massey University, Palmerston North, New Zealand.
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MeSH Terms
Descriptor/Qualifier:
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*
Chemical
Reg. No./Substance:
0/Cross-Linking Reagents; EC 3.2.1.26/beta-Fructofuranosidase

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


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