A precise swaying map for how promiscuous cellobiose-2-epimerase operate bi-reaction

Yinghui Feng, Xiaomei Lyu, Yalong Cong, Tingwei Miao, Bohuan Fang, Chuanxi Zhang, Qiang Shen, Melissa Matthews, Andrew J. Fisher, John Z.H. Zhang, Lujia Zhang, Ruijin Yang

Research output: Contribution to journalArticlepeer-review


Promiscuous enzymes play a crucial role in organism survival and new reaction mining. However, comprehensive mapping of the catalytic and regulatory mechanisms hasn't been well studied due to the characteristic complexity. The cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus (CsCE) with complex epimerization and isomerization was chosen to comprehensively investigate the promiscuous mechanisms. Here, the catalytic frame of ring-opening, cis-enediol mediated catalysis and ring-closing was firstly determined. To map the full view of promiscuous CE, the structure of CsCE complex with the isomerized product glucopyranosyl-β1,4-fructose was determined. Combined with computational calculation, the promiscuity was proved a precise cooperation of the double subsites, loop rearrangement, and intermediate swaying. The flexible loop was like a gear, whose structural reshaping regulates the sway of the intermediates between the two subsites of H377-H188 and H377-H247, and thus regulates the catalytic directions. The different protonated states of cis-enediol intermediate catalyzed by H188 were the key point for the catalysis. The promiscuous enzyme tends to utilize all elements at hand to carry out the promiscuous functions.

Original languageEnglish (US)
Article number127093
JournalInternational Journal of Biological Macromolecules
StatePublished - Dec 31 2023


  • Cellobiose 2-epimerase
  • Cis-enediol intermediate
  • Promiscuous enzyme

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology


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