AI Article Synopsis
- Glutamate dehydrogenases (GDH) are crucial enzymes that connect protein and carbohydrate metabolism, and researchers have engineered novel GDHs through motif reassembly and mutations to enhance their stability and activity in nonaqueous environments.
- Molecular dynamics simulations were used to analyze the structural compatibility and dynamic interactions of these new enzymes, while multipoint mutations were found to significantly boost their catalytic activity for unconventional substrates.
- The study demonstrated that an increased presence of hydrogen-bonded salt bridges supports enzyme stability, and experimental results showed a 1.78-fold increase in the activity of a specific engineered enzyme, Aa05, when used with an ionic liquid, highlighting an effective strategy for designing robust dehydrogenases in challenging conditions.
Article Abstract
Glutamate dehydrogenases (GDH) serve as the key regulated enzyme that links protein and carbohydrate metabolism. Combined with motif reassembly and mutation, novel GDHs were designed. Motif reassembly of thermophilic GDH and malate dehydrogenase aims to overcome stability and activity tradeoff in nonaqueous systems. Structural compatibility and dynamic cooperation of the designed AaDHs were studied by molecular dynamics simulation. Furthermore, multipoint mutations improved its catalytic activity for unnatural substrates. Amino acid interaction network analysis indicated that the high density of hydrogen-bonded salt bridges is beneficial to the stability. Finally, the experimental verification determines the kinetics of AaDHs in a nonaqueous system. The activity of Aa05 was increased by 1.78-fold with ionic liquid [EMIM]BF. This study presents the strategy of a combination of rigid motif assembly and mutations of active sites for robust dehydrogenases with high activity in the nonaqueous system, which overcomes the activity-stability tradeoff effect.
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| http://dx.doi.org/10.1021/acs.jafc.4c02995 | DOI Listing |
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