2024-03-29T06:26:20Zhttps://repositori.uji.es/oai/requestoai:repositori.uji.es:10234/1520052023-06-28T08:03:06Zcom_10234_7013com_10234_9col_10234_8638
00925njm 22002777a 4500
dc
Sánchez Moreno, Israel
author
Bordes, Isabel
author
Castillo, Raquel
author
Ruiz-Pernía, José Javier
author
Moliner, Vicent
author
García Junceda, Eduardo
author
2015-11-24
Dihydroxyacetone (DHA) kinase from Citrobacter freundii provides an easy entry for the preparation of DHA phosphate; a very important C3 building block in nature. To modify the phosphoryl donor specificity of this enzyme from ATP to inorganic polyphosphate (poly-P); a directed evolution program has been initiated. In the first cycle of evolution, the native enzyme was subjected to one round of error-prone PCR (EP-PCR) followed directly (without selection) by a round of DNA shuffling. Although the wild-type DHAK did not show activity with poly-P, after screening, sixteen mutant clones showed an activity with poly-phosphate as phosphoryl donor statistically significant. The most active mutant presented a single mutation (Glu526Lys) located in a flexible loop near of the active center. Interestingly, our theoretical studies, based on molecular dynamics simulations and hybrid Quantum Mechanics/Molecular Mechanics (QM/MM) optimizations, suggest that this mutation has an effect on the binding of the poly-P favoring a more adequate position in the active center for the reaction to take place.
SÁNCHEZ-MORENO, Israel, et al. Tuning the Phosphoryl Donor Specificity of Dihydroxyacetone Kinase from ATP to Inorganic Polyphosphate. An Insight from Computational Studies. International journal of molecular sciences, 2015, vol. 16, no 11, p. 27835-27849.
1661-6596
1422-0067
http://hdl.handle.net/10234/152005
http://dx.doi.org/10.3390/ijms161126073
Biocatalysis
Computational chemistry
DHAP-dependent aldolases
Dihydroxyacetone kinase
Enzyme directed evolution
Quantum mechanics/molecular mechanics
Tuning the Phosphoryl Donor Specificity of Dihydroxyacetone Kinase from ATP to Inorganic Polyphosphate. An Insight from Computational Studies