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United States Department of Agriculture

Agricultural Research Service

Research Project: INDUSTRIALLY ROBUST ENZYMES AND MICROORGANISMS FOR PRODUCTION OF SUGARS AND ETHANOL FROM AGRICULTURAL BIOMASS Title: Active-Site Models of Riboflavin Synthase

Authors
item Jordan, Douglas
item Calabrese, Joseph - DUPONT
item Liao, Der-Ing - DUPONT
item Wawrzak, Zdzislaw - DND-CAT SYNCHROTRON RES
item Zheng, Ya-Jun - DUPONT

Submitted to: Meeting Proceedings
Publication Type: Proceedings
Publication Acceptance Date: October 1, 2005
Publication Date: December 1, 2005
Citation: Jordan, D.B., Calabrese, J.C., Liao, D., Wawrzak, Z., Zheng, Y. 2005. Active-site models of riboflavin synthase. In: Nishino, T., Miura, R., Tanokura, M., Fukui, K., editors. Flavins and Flavoproteins 2005. Tokyo: ARchiTect, Inc. p. 737-742.

Technical Abstract: Riboflavin synthase (RS) catalyses the dismutation of two molecules of 6,7-dimethyl-8-(1´-D-ribityl)-lumazine (DMRL) to yield riboflavin and 4-ribitylamino-5-amino-2,6-dihydroxypyrimidine (RAADP). One four-carbon unit is transferred from the donor DMRL, which becomes RAADP, to the acceptor DMRL, which becomes riboflavin. RS is a homotrimer of 23 kDa subunits with each subunit consisting of two similar beta barrels and a C-terminal alpha helix. Active sites are formed at the interface between an N barrel (beta barrel belonging to the N terminus) of one subunit and a C barrel (beta barrel belonging to the C terminus) of another subunit. Only one active site of the three in the trimer can be formed at a time. We report the X-ray structure of the Escherichia coli RS in complex with riboflavin. Five riboflavins reside in the homotrimer, with one bound to each beta barrel except the C barrel that is part of the formed active site. The N barrel of the formed active site is occupied by riboflavin. This result substantiates models of the Michaelis complex and the pentacyclic reaction intermediate, which are based on the acceptor DMRL molecule belonging to the N barrel and the donor DMRL molecule belonging to the C barrel.

Last Modified: 7/31/2014
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