Relationship between Escherichia coli AppA phytase's thermostability and salt bridges

Fei, Baojin; Xu, Hui; Zhang, Feiwei; Li, Xinran; Ma, Shuhan; Cao, Yu; Xie, Jinlu; Qiao, Dan; Cao, Yi

doi:10.1016/j.jbiosc.2012.12.010

Cited by 21 publications

(16 citation statements)

References 33 publications

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“…The residual activities of the wild-type AppA phytase and C-lose mutant were 31.42% and 70.49%, respectively, after being heated at 80°C for 10 min [22]. Fei et al also constructed a salt bridge addition mutant Q307D by site-directed mutagenesis, which showed 9.15% thermostability enhancement than the wild-type after being heated at 80°C for 10 min [23]. In this study, compared with the wild-type, the thermal stability of the modified enzyme has been greatly improved.…”

Section: Analysis Of Amino Acids In Different Mutation Librariesmentioning

confidence: 99%

“…Based on the crystal structure and rational cascade methods, Fei et al constructed a series of engineered phytases, and the mutants showed a great increase of heat resistance compared to the wild-type [19]. Furthermore, under the guidance of 3D structural information, analysis and calculation of some types of molecular bonds, such as hydrogen bonds [22], salt bridges [23] and disulfide bonds [24], have also been utilized to improve the heat and pH resistance of phytase. In other examples, glycosylation [25] and modification of α/β structure domains [26] were also successfully applied to improve the heat resistance of the protein.…”

Section: Introductionmentioning

confidence: 99%

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Evolution of E. coli Phytase for Increased Thermostability Guided by Rational Parameters

Gai

et al. 2019

Journal of Microbiology and Biotechnology

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Phytases are enzymes that can hydrolyze phytate and its salts into inositol and phosphoric acid, and have been utilized to increase the availability of nutrients in animal feed and mitigate environmental pollution. However, the enzymes' low thermostability has limited their application during the feed palletization process. In this study, a combination of B-value calculation and protein surface engineering was applied to rationally evolve the heat stability of Escherichia coli phytase. After systematic alignment and mining for homologs of the original phytase from the histidine acid phosphatase family, the two models 1DKL and 1DKQ were chosen and used to identify the B-values and spatial distribution of key amino acid residues. Consequently, thirteen potential amino acid mutation sites were obtained and categorized into six domains to construct mutant libraries. After five rounds of iterative mutation screening, the thermophilic phytase mutant P56214 was finally yielded. Compared with the wild-type, the residual enzyme activity of the mutant increased from 20% to 75% after incubation at 90°C for 5 min. Compared with traditional methods, the rational engineering approach used in this study reduces the screening workload and provides a reference for future applications of phytases as green catalysts.

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Section: Analysis Of Amino Acids In Different Mutation Librariesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Evolution of E. coli Phytase for Increased Thermostability Guided by Rational Parameters

Gai

et al. 2019

Journal of Microbiology and Biotechnology

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“…Figure 5 indicates the trends of salt bridge formation and breakage in case of all considered mutants as well as the original protein. In general, surface salt bridges have been shown to contribute to protein stability at high temperatures due to decreased desolvation penalties of charged groups (Fei et al, 2013).…”

Section: Low Solvent Accessible Surface Area (Sasa) Of Hydrophobic Mumentioning

confidence: 99%

<i>In-silico</i> Rational Protein Engineering and Design Approach to Improve Thermostability of a Haloalkane Dehalogenase Enzyme

Satpathy¹,

Konkimalla²,

Ratha³

2015

American Journal of Bioinformatics

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Thermostability of enzymes is a major prerequisite for use in industrial enzymology. There are as such no simple general principles for achieving thermostability in case of enzymes as many factors are required to fulfil for different enzymes. The present study describes computational methods to design thermostable haloalkane dehalogenase enzyme using the crystal structure available Protein Data Bank (PDB ID: 1EDE). In in silico design strategy rule-based approaches such as disulfide bond geometry, new hydrophobic pocket design, new salt bridge construction and multiple mutations (combination of the above approaches) were introduced to the original enzyme. After each design strategy the functional effect was confirmed in terms of enzyme substrate binding by molecular docking using Autodock vina tool. Best design strategy was evaluated by comparative molecular dynamics simulation applying simulated annealing method at 8 ns using GROMACS tool. The surface hydrophobicity which is the key factor for thermostability in haloalkane dehalogenase was obtained from the simulation result. Upon optimizing the parameters, thermostability of mutant enzyme under consideration was also confirmed by the 5 ns molecular dynamics simulation at 400, 500 and 600 K.

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“…In monogastric animals, phosphorus is largely unavailable in phytic acid due to the absence of phytase in their digestive system. This leads to the elimination of phytic acid via stool, which consequently results in soil pollution and eutrophication of water by phosphates (Fei et al, 2013;Ma et al, 2011;Vats et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Optimized production of phytase by solid-state fermentation using triticale as substrate and source of inducer

Neira-Vielma¹,

Cristobal²,

Anna³

et al. 2018

Afr. J. Biotechnol.

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This study was carried out to evaluate the process of phytase production by Aspergillus niger in solidstate fermentation (SSF) using triticale waste. A waste that currently has no use was reported for this biotechnological process, and is of high impact due to the null use. The process was carried out using an additive free medium, supplemented with only one nitrogen source. Under these conditions, phytase activity of 7.45 U/g dry substrate (DS) was obtained. The process was optimized using different additives such as dextrose, lactose, Tween 80 and potassium chloride. For fermentation maximization, two experimental designs were used: 1) Plackett-Burman design (PBD) and 2) the Box-Behnken design (BBD). PBD was used to evaluate the effect of related variables on the production of phytase, as well as their level of significance in the process, while BBD was used for optimal conditions determination. The process was conducted with Petri dishes and a maximum enzyme activity of 25.8 IU/g DS was obtained. Subsequently, SSF was carried out in a tray to increase the amount of fermented substrate and phytase activity of 23.63 IU/g DS was obtained. The results of this study suggest a minimal decrease (8.4%) in enzyme production with scaling.

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Relationship between Escherichia coli AppA phytase's thermostability and salt bridges

Cited by 21 publications

References 33 publications

Evolution of E. coli Phytase for Increased Thermostability Guided by Rational Parameters

Evolution of E. coli Phytase for Increased Thermostability Guided by Rational Parameters

<i>In-silico</i> Rational Protein Engineering and Design Approach to Improve Thermostability of a Haloalkane Dehalogenase Enzyme

Optimized production of phytase by solid-state fermentation using triticale as substrate and source of inducer

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