Alternative Epimerization in C7N-Aminocyclitol Biosynthesis Is Catalyzed by ValD, A Large Protein of the Vicinal Oxygen Chelate Superfamily

作者：Xu, Hui; Zhang, Yirong; Yang, Jongtae; Mahmud, Taifo^*; Bai, Linquan; Deng, Zixin

来源：Chemistry & Biology, 2009, 16(5): 567-576.

DOI：10.1016/j.chembiol.2009.04.006

• GLUCOSIDASE INHIBITOR ACARBOSE
• HYGROSCOPICUS VAR. LIMONEUS
• GENE-CLUSTER
• VALIDAMYCIN-A
• EVOLUTIONARY RELATIONSHIPS
• RESISTANCE PROTEIN
• CRYSTAL-STRUCTURE
• ACTINOPLANES SP
• MOIETY
• 2-EPI-5-EPI-VALIOLONE

Gene valD, encodes a large vicinal oxygen chelate (VOC) superfamily protein, has been identified in the validamycin biosynthetic gene cluster. Inactivation of valD significantly reduced validamycin A production, which was fully restored with the full-length valD and partially restored with either N-terminal or C-terminal half by complementation. Heterologously expressed ValD catalyzed the epimerization of 2-epi-5-epi-valiolone to 5-epi-valiolone. This metalloenzyme is a homodimer with a metal ion-binding ratio of 0.73 mol/mole protein toward Fe2+, Mn2+, Ni2+, and Zn2+. Individual and combined site-directed mutations of eight putative active site residues revealed that the N-terminal H44/E107 and the C-terminal H315/E366 are more critical for the activity than the internal H130, E183, H229, and E291. Our data have established ValD as one of the largest proteins of the VOC superfamily, catalyzing an alternative epimerization for C7N-aminocyclitol biosynthesis.

• 出版日期2009-5-29
• 单位上海交通大学