Cloning, expression and characterization of an aryl-alcohol dehydrogenase from the white-rot fungus Phanerochaete chrysosporium strain BKM-F-1767

BMC Microbiology

Research Article

Dong-Dong Yang¹  Jean Marie François¹  Gustavo M de Billerbeck² 

[1] Université de Toulouse; INSA, UPS, INP; LISBP, 135 Avenue de Rangueil, F-31077, Toulouse, France;INRA, UMR792, Ingénierie des Systèmes Biologiques et des Procédés, F-31400, Toulouse, France;CNRS, UMR5504, F-31400, Toulouse, France;Université de Toulouse; INSA, UPS, INP; LISBP, 135 Avenue de Rangueil, F-31077, Toulouse, France;INRA, UMR792, Ingénierie des Systèmes Biologiques et des Procédés, F-31400, Toulouse, France;CNRS, UMR5504, F-31400, Toulouse, France;INP-ENSAT, Avenue de l’Agrobiopole, F-31326, Castanet-Tolosan Cedex, France;

关键词: AAD;    Aryl-alcohol dehydrogenase;    Lignocellulosic hydrolysates;    Lignin;    Flavours;    Fragrances;    Phanerochaete chrysosporium;   

DOI  :  10.1186/1471-2180-12-126

 received in 2012-02-01, accepted in 2012-04-16,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundThe white-rot fungus Phanerochaete chrysosporium is among the small group of fungi that can degrade lignin to carbon dioxide while leaving the crystalline cellulose untouched. The efficient lignin oxidation system of this fungus requires cyclic redox reactions involving the reduction of aryl-aldehydes to the corresponding alcohols by aryl-alcohol dehydrogenase. However, the biochemical properties of this enzyme have not been extensively studied. These are of most interest for the design of metabolic engineering/synthetic biology strategies in the field of biotechnological applications of this enzyme.ResultsWe report here the cloning of an aryl-alcohol dehydrogenase cDNA from the white-rot fungus Phanerochaete chrysosporium, its expression in Escherichia coli and the biochemical characterization of the encoded GST and His6 tagged protein. The purified recombinant enzyme showed optimal activity at 37°C and at pH 6.4 for the reduction of aryl- and linear aldehydes with NADPH as coenzyme. NADH could also be the electron donor, while having a higher Km (220 μM) compared to that of NADPH (39 μM). The purified recombinant enzyme was found to be active in the reduction of more than 20 different aryl- and linear aldehydes showing highest specificity for mono- and dimethoxylated Benzaldehyde at positions 3, 4, 3,4 and 3,5. The enzyme was also capable of oxidizing aryl-alcohols with NADP + at 30°C and an optimum pH of 10.3 but with 15 to 100-fold lower catalytic efficiency than for the reduction reaction.ConclusionsIn this work, we have characterized the biochemical properties of an aryl-alcohol dehydrogenase from the white-rot fungus Phanerochaete chrysosporium. We show that this enzyme functions in the reductive sense under physiological conditions and that it displays relatively large substrate specificity with highest activity towards the natural compound Veratraldehyde.

【 授权许可】

Unknown   
© Yang et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.