Abstract

The ERY4 laccase gene from Pleurotus eryngii was expressed in Saccharomyces cerevisiae and the recombinant laccase resulted to be not biologically active. This gene was thus modified to obtain chimerical enzymes derived from the substitution of N-, C- and both N-and C-terminal regions with the corresponding regions of Ery3 laccase, another laccase isoform of P. eryngii. The chimerical isoform named 4NC3, derived from the substitution of both N-and C-terminal regions, showed the best performances in terms of enzymatic activities, affinities for different substrates and stability at a broad range of temperatures and pHs. The chimerical 4NC3 laccase isoform was displayed on the cell surface of S. cerevisiae using the N-terminal fusion with either the Pir2 or the Flo1 S. cerevisiae proteins as anchor attachment sequence. Immunofluorescence microscopy and Western blot analyses confirmed the localization of 4NC3 on the yeast cell surface. The enzyme activity on specific laccase substrates revealed that 4NC3 laccase was immobilized in active form on the cell surface. To our knowledge, this is the first example of expression of a chimerical fungal laccase by yeast cell display.

Autore Pugliese

• Rampino Patrizia , Perrotta Carla

Tutti gli autori

• Bleve G. , Lezzi C. , Spagnolo S. , Rampino P. , Perrotta C. , Mita G. , Grieco F.

Titolo volume/Rivista

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

Anno di pubblicazione

2014

ISSN

0273-2289

ISBN

Non Disponibile

Numero di citazioni Wos

14

Ultimo Aggiornamento Citazioni

28/04/2018

Numero di citazioni Scopus

16

Ultimo Aggiornamento Citazioni

28/04/2018

Settori ERC

Non Disponibile

Codici ASJC

Non Disponibile