Patent ID: 6184015
Filing Date: 2001-02-06
Classification: C11D,C12N,C12Y

Abstract:
A method of constructing a variant of a parent Coprinus-like laccase having laccase activity and having one or more of the following alterations:a. altered oxidation potentialb. altered pH optimumc. altered mediator efficiencyd. altered O.sub.2 /OH.sup.- -pathwayas compared to said parent Coprinus-like laccase,said method comprising:I) modeling the parent Coprinus-like laccase on the three-dimensional amino acid structure of Coprinus cinereus laccase, as defined by the atomic coordinates in Table 1, to produce a three-dimensional modeled structure of the parent Coprinus-like laccase,ii) identifying, in the three-dimensional modeled structure obtained in step (I), a structural part of the parent Coprinus-like laccase which is predicted to be responsible for differences in oxidation potential, pH optimum, mediator efficiency, or O.sub.2 /OH.sup.- -pathway between the parent Coprinus-like laccase and the Coprinus cinereus laccase, andiii) modifying the part of the parent Coprinus-like laccase identified in step (ii) to produce a variant of the parent Coprinus-like laccase, said variant having laccase activity and altered oxidation potential and/or altered pH optimum and/or altered mediator efficiency and/or an altered O.sub.2 /OH.sup.- -pathway as compared to said parent Coprinus-like laccase;wherein said parent Coprinus-like laccase has laccase enzymatic activity and has an amino acid sequence at least 50% homologous to SEQ ID NO: 1 when homology is measured by GAP Version 8.1, using a gap weight of 3.0 and a length weight of 0.1.