作者:Feng Xu、Elizabeth J. Golightly、Claus C. Fuglsang、Palle Schneider、Kyle R. Duke、Loi Lam、Søren Christensen、Kimberly M. Brown、Christel T. Jørgensen、Stephen H. Brown
DOI:10.1046/j.1432-1327.2001.01982.x
日期:2001.2
A Microdochium nivale carbohydrate:acceptor oxidoreductase was purified, cloned, heterologously expressed, and characterized. The gene encoding the protein showed one intron, and the ORF showed a sequence with low homology (≤ 25% identity or 65% similarity) to other known flavin‐containing carbohydrate oxidases. The maturation of the protein required the cleavage of a tetrameric propeptide in addition to an 18 amino‐acid signal peptide. The enzyme was found to have a relative molecular mass of 55 000 Da, an isoelectric point of 9, and one FAD per protein. It could oxidize mono‐, oligo‐, or polymeric saccharides, and transfer their electrons to O2 or other acceptors. When d‐glucose served as electron‐donating substrate, an activity of 2 s−1 was observed at pH 5.5 and 23 °C. Among various oligosaccharides, the enzyme preferred tetrameric dextrins, indicating a favorable interaction of four linked glucose units with the substrate pocket. The unique structure and ability of oxidizing oligo/polymeric saccharides suggest a promising prospect of this enzyme for various industrial/medicinal applications.