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. 2012 Mar 17;3(2):123–131. doi: 10.1007/s13238-012-2026-5

Crystal structures of d-psicose 3-epimerase from Clostridium cellulolyticum H10 and its complex with ketohexose sugars

Hsiu-Chien Chan 1, Yueming Zhu 1, Yumei Hu 1, Tzu-Ping Ko 2, Chun-Hsiang Huang 1, Feifei Ren 1, Chun-Chi Chen 3, Yanhe Ma 1, Rey-Ting Guo 1,✉, Yuanxia Sun 1,✉
PMCID: PMC4875416  PMID: 22426981

Abstract

d-Psicose 3-epimerase (DPEase) is demonstrated to be useful in the bioproduction of d-psicose, a rare hexose sugar, from d-fructose, found plenty in nature. Clostridium cellulolyticum H10 has recently been identified as a DPEase that can epimerize d-fructose to yield d-psicose with a much higher conversion rate when compared with the conventionally used DTEase. In this study, the crystal structure of the C. cellulolyticum DPEase was determined. The enzyme assembles into a tetramer and each subunit shows a (β/α)8 TIM barrel fold with a Mn2+ metal ion in the active site. Additional crystal structures of the enzyme in complex with substrates/products (d-psicose, d-fructose, d-tagatose and d-sorbose) were also determined. From the complex structures of C. cellulolyticum DPEase with d-psicose and d-fructose, the enzyme has much more interactions with d-psicose than d-fructose by forming more hydrogen bonds between the substrate and the active site residues. Accordingly, based on these ketohexose-bound complex structures, a C3-O3 proton-exchange mechanism for the conversion between d-psicose and d-fructose is proposed here. These results provide a clear idea for the deprotonation/protonation roles of E150 and E244 in catalysis.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s13238-012-2026-5 and is accessible for authorized users.

Keywords: d-psicose 3-epimerase, ketohexose, complex structure

Electronic supplementary material

13238_2012_2026_MOESM1_ESM.pdf (1.2MB, pdf)

Supplementary material, approximately 1.17 MB.

Footnotes

These authors contributed equally to the work.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s13238-012-2026-5 and is accessible for authorized users.

Contributor Information

Rey-Ting Guo, Email: guo_rt@tib.cas.cn.

Yuanxia Sun, Email: sun_yx@tib.cas.cn.

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13238_2012_2026_MOESM1_ESM.pdf (1.2MB, pdf)

Supplementary material, approximately 1.17 MB.

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