The improvement of stability, activity, and substrate promiscuity of glycerol dehydrogenase substituted by divalent metal ions

Wang, Shizhen; Wang, Jing; Zhou, Xiaofen; Guo, Yingxia; Fang, Baishan

doi:10.1007/s12257-013-0125-7

Cited by 16 publications

(10 citation statements)

References 20 publications

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“…The results indicated that the addition of Mn 2+ to the reaction solution decreased the activity of GDH. The activity of the apoenzyme after EDTA treatment was nearly undetectable, which confirmed the activity enhancement by metal ion substitution [21] .…”

Section: Methodssupporting

confidence: 65%

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Mechanistic Study of Manganese-Substituted Glycerol Dehydrogenase Using a Kinetic and Thermodynamic Analysis

et al. 2014

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Mechanistic insights regarding the activity enhancement of dehydrogenase by metal ion substitution were investigated by a simple method using a kinetic and thermodynamic analysis. By profiling the binding energy of both the substrate and product, the metal ion's role in catalysis enhancement was revealed. Glycerol dehydrogenase (GDH) from Klebsiella pneumoniae sp., which demonstrated an improvement in activity by the substitution of a zinc ion with a manganese ion, was used as a model for the mechanistic study of metal ion substitution. A kinetic model based on an ordered Bi-Bi mechanism was proposed considering the noncompetitive product inhibition of dihydroxyacetone (DHA) and the competitive product inhibition of NADH. By obtaining preliminary kinetic parameters of substrate and product inhibition, the number of estimated parameters was reduced from 10 to 4 for a nonlinear regression-based kinetic parameter estimation. The simulated values of time-concentration curves fit the experimental values well, with an average relative error of 11.5% and 12.7% for Mn-GDH and GDH, respectively. A comparison of the binding energy of enzyme ternary complex for Mn-GDH and GDH derived from kinetic parameters indicated that metal ion substitution accelerated the release of dioxyacetone. The metal ion's role in catalysis enhancement was explicated.

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Section: Methodssupporting

confidence: 65%

“…is a zinc-dependent metalloenzyme [20] . It has been previously shown that Mn 2+ substituted GDH exhibits improved activity and thermostability [21] . In this paper, mechanistic insights of activity improvement are studied based on kinetic and thermodynamic analysis.…”

Section: Introductionmentioning

confidence: 99%

Mechanistic Study of Manganese-Substituted Glycerol Dehydrogenase Using a Kinetic and Thermodynamic Analysis

et al. 2014

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“…Compared to the original activity of GDH (3.17 U/mL), the activities of GDH modified by Mg 2+ , Ba 2+ , and Mn 2+ were improved by 14.9-, 11.3-, and 12.4-folds after optimization, respectively [ 160 ]. The GDH catalytic zinc ion substitution by other divalent metal ions, Mn 2+ and Mg 2+ , had increased the activities of GDH; however, their thermostability and catalytic promiscuity have not yet been studied [ 161 ] (Table 7 ). The mechanism of the metal ion’s role in the catalytic enhancement was explained [ 12 ].…”

Section: Glycerol Dehydrogenasementioning

confidence: 99%

Key enzymes catalyzing glycerol to 1,3-propanediol

Jiang

Wang

et al. 2016

Biotechnol Biofuels

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Biodiesel can replace petroleum diesel as it is produced from animal fats and vegetable oils, and it produces about 10 % (w/w) glycerol, which is a promising new industrial microbial carbon, as a major by-product. One of the most potential applications of glycerol is its biotransformation to high value chemicals such as 1,3-propanediol (1,3-PD), dihydroxyacetone (DHA), succinic acid, etc., through microbial fermentation. Glycerol dehydratase, 1,3-propanediol dehydrogenase (1,3-propanediol-oxydoreductase), and glycerol dehydrogenase, which were encoded, respectively, by dhaB, dhaT, and dhaD and with DHA kinase are encompassed by the dha regulon, are the three key enzymes in glycerol bioconversion into 1,3-PD and DHA, and these are discussed in this review article. The summary of the main research direction of these three key enzyme and methods of glycerol bioconversion into 1,3-PD and DHA indicates their potential application in future enzymatic research and industrial production, especially in biodiesel industry.

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“…Structure study of GDH from Bacillus stearothermophilus demonstrated that divalent metal ion, Zn 2+ was required for the multimer formation and enzymatic catalysis of GDH 21 . The natural GDH-bound Zn 2+ were substituted with several divalent metal ions and the enzyme activity was significantly altered 22 23 . Therefore we selected six divalent metal ions (Cu 2+ , Ni 2+ , Zn 2+ , Mn 2+ , Mg 2+ and Ca 2+ ) to investigate the effects of metal ions on GDH activity.…”

Section: Resultsmentioning

confidence: 99%

Bioinspired Immobilization of Glycerol Dehydrogenase by Metal Ion-Chelated Polyethyleneimines as Artificial Polypeptides

Zhang

Ren

Wang

et al. 2016

Sci Rep

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In this study, a novel, simple and generally applicable strategy for multimeric oxidoreductase immobilization with multi-levels interactions was developed and involved activity and stability enhancements. Linear polyethyleneimines (PEIs) are flexible cationic polymers with molecular weights that span a wide range and are suitable biomimic polypeptides for biocompatible frameworks for enzyme immobilization. Metal ion-chelated linear PEIs were applied as a heterofunctional framework for glycerol dehydrogenase (GDH) immobilization by hydrogen bonds, electrostatic forces and coordination bonds interactions. Nanoparticles with diameters from 250–650 nm were prepared that exhibited a 1.4-fold enhancement catalytic efficiency. Importantly, the half-life of the immobilized GDH was enhanced by 5.6-folds in aqueous phase at 85 °C. A mechanistic illustration of the formation of multi-level interactions in the PEI-metal-GDH complex was proposed based on morphological and functional studies of the immobilized enzyme. This generally applicable strategy offers a potential technique for multimeric enzyme immobilization with the advantages of low cost, easy operation, high activity reservation and high stability.

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The improvement of stability, activity, and substrate promiscuity of glycerol dehydrogenase substituted by divalent metal ions

Cited by 16 publications

References 20 publications

Mechanistic Study of Manganese-Substituted Glycerol Dehydrogenase Using a Kinetic and Thermodynamic Analysis

Mechanistic Study of Manganese-Substituted Glycerol Dehydrogenase Using a Kinetic and Thermodynamic Analysis

Key enzymes catalyzing glycerol to 1,3-propanediol

Bioinspired Immobilization of Glycerol Dehydrogenase by Metal Ion-Chelated Polyethyleneimines as Artificial Polypeptides

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