مقاله Engineering and production of laccase from Trametes versicolor in the yeast Yarrowia lipolytica : تی پی بین

خلاصه مقاله

The lcc1 gene coding for the laccase from Trametes versicolor DSM11269 was cloned into the genome of Yarrowia lipolytica using either single or multiple integration sites. The levels of the recombinant laccase activity secreted in the culture media were 0.25 and 1 U ml⁻¹ for single and multiple integrations, respectively. The strain with a single integration was successfully used to express variant libraries which were screened on ABTS substrate. The strain encoding the double mutant L185P/Q214K (rM4A) showed a sixfold enhancement in secreted enzyme activity. The catalytic efficiency of the purified rM-4A laccase was respectively increased 2.4- and 2.8-fold towards ABTS and 2,6-dimethoxyphenol, compared to the rWT. Culture supernatants containing either rWT or rM-4A catalyzed the almost complete decolorization of an Amaranth solution (70 nM s⁻¹). Taken together, our results open new perspectives for the use of Y. lipolytica as a molecular evolution platform to engineer laccases with improved properties.

نویسندگان

Monnat Theerachat [ Biological Science Program, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand ]

Stéphane Emond [ Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK ]

Emmanuelle Cambon [ Université de Toulouse, INSA, UPS, INP, LISBP, 135 Avenue de Rangueil, F-31077 Toulouse, France ]

Florence Bordes [ Université de Toulouse, INSA, UPS, INP, LISBP, 135 Avenue de Rangueil, F-31077 Toulouse, France ]

Alain Marty [ Université de Toulouse, INSA, UPS, INP, LISBP, 135 Avenue de Rangueil, F-31077 Toulouse, France ]

کلمات کلیدی

Laccase Trametes versicolor Decolorization Yarrowia lipolytica

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Bioresource Technology

The journal's aim is to advance and disseminate knowledge in all the related areas of biomass, biological waste treatment, bioenergy, biotransformations and bioresource systems analysis, and technologies associated with conversion or production.

حوزه های تحت پوشش : Bioresource Technology