Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.10.3.2 (laccase)
 4,656 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Abstract The fungal pathogen Botrytis cinerea is capable of developing on a wide variety of host plants that differ greatly in their pH values and biochemical defences. To evaluate whether the pH of the host tissue can regulate the production of pathogenicity factors by this fungus, we examined the ability of two isolates of B. cinerea that originated from different plant species to secrete putative virulence elements on synthetic media buffered at pH 2.0 to pH 7.0. Even though differing in the intensity of their responses, both isolates reacted similarly to their ambient pH. The production of extracellular polysaccharides and oxalic acid was detectable above pH 4.0 and pH 5.0 respectively. Conversely, the production of aspartic acid proteases could only be seen between pH 3.0 and 4.0. Finally, the secretion of polygalacturonase and laccase activity was found to exhibit two maxima, one around pH 3.1 and one around pH 6.0. Thus, pathogenicity factor production was found to be minimal between pH 4.5 and 5.5 and a different set of factors was produced at pH 3.1 and 6.0, two values that were found to correspond respectively to the average host fruit and leaf pH. These results demonstrate that ambient pH differentially regulates the synthesis of pathogenicity factors by Botrytis and may act as a novel regulatory element to assist this fungus in tuning its virulence machinery to the composition of its host tissue.
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PMID:Differential regulation by ambient pH of putative virulence factor secretion by the phytopathogenic fungus Botrytis cinerea. 1971 67

Structure-function relationships underlying laccases properties are very limited that makes these enzymes interesting for protein engineering approaches. Therefore in the current study, a thermostable laccase that was isolated from Bacillus sp. HR03 with the ability of bilirubin oxidation besides its laccase and tyrosinase activity is used. The extensive application of this enzyme is limited by its low expression level in Escherichia coli. Based on sequence alignments and structural studies, three single amino acid substitutions, D500G, D500E, D500S and a glycine insertion, are introduced using site-directed mutagenesis to evaluate the role of Asp(500) located in the C-terminal segment close to the T1 copper center. Substitution of aspartic acid with less sterically hindered, conserved residue such as glycine increase kcat (2.3 fold) and total activity (7.3 fold) which is accompanied by a significant increase in the expression level up to 3 fold. Biochemical characterization and structural studies using far-UV CD and fluorescence spectroscopy reveal the importance of C-terminal copper-binding loop in the laccase functional expression and catalytic efficiency. Kinetic characterization of the purified mutants toward 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), syringaldazine (SGZ) and bilirubin, shows that substrate specificity is left unchanged.
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PMID:Enhancement of catalysis and functional expression of a bacterial laccase by single amino acid replacement. 2370 61