Thermally stable and acidic pH tolerant mutant phytases with high catalytic efficiency from Yersinia intermedia for potential application in feed industries

Heat- and pH-stable phytase efficiently hydrolyzes phytic acid. In this research, heat- and pH-stable mutant phytases, T 83 R, L 287 R, and T 83 R/L 287 R were generated by site-directed mutagenesis from Yersinia intermedia . After the induction and expression of recombinant wild-type and mutant phy...

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Bibliographic Details
Published in:	Environmental science and pollution research international Vol. 29; no. 22; pp. 33713 - 33724
Main Authors:	Abbasi Kheirabadi, Marjan, Saffar, Behnaz, Hemmati, Roohullah, Mortazavi, Mojtaba
Format:	Journal Article
Language:	English
Published:	Berlin/Heidelberg Springer Berlin Heidelberg 01-05-2022 Springer Nature B.V
Subjects:	6-Phytase - chemistry Affinity chromatography Aquatic Pollution Atmospheric Protection/Air Quality Control/Air Pollution Bioavailability E coli Earth and Environmental Science Ecotoxicology Environment Environmental Chemistry Environmental Health Environmental science Enzyme Stability Enzymes Escherichia coli - metabolism Feed industry Hydrogen-Ion Concentration Minerals Mutants Nickel pH effects Phytase Phytic acid Research Article Site-directed mutagenesis Soil pollution Soil water Spectroscopy Thermal stability Waste Water Technology Water Management Water pollution Water Pollution Control Yersinia - chemistry Yersinia intermedia Feed industries Mutagenesis Environmental pollution Protein purification Phytic acid Thermal stability
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Summary:	Heat- and pH-stable phytase efficiently hydrolyzes phytic acid. In this research, heat- and pH-stable mutant phytases, T 83 R, L 287 R, and T 83 R/L 287 R were generated by site-directed mutagenesis from Yersinia intermedia . After the induction and expression of recombinant wild-type and mutant phytases in E. coli BL21, the enzymes were purified using nickel sepharose affinity chromatography, and characterized kinetically and thermodynamically using spectroscopy methods. The mutants showed optimum activity at pH 5.15 and 55–61 ° C. The catalytic efficiencies of T 83 R, L 287 R, T 83 R/L 287 R, and wild-type phytases were calculated to be 2941, 29346, 4906, and 6917 mmol/L −1 s −1 , respectively. Moreover, after the incubation of T 83 R, L 287 R, wild-type, and T 83 R/ L 287 R phytases at 100 ° C for 1 h, the enzymes retained 22, 5, 4, and 2% of their initial activities, respectively. In addition, T 83 R, T 83 R/L 287 R, L 287 R, and wild-type phytases retained 82, 44, 16 as well as 11% of their initial activities after 1 h at pH 5.15, respectively. Among these mutants, T 83 R mutant showed 18% increase in thermal stability, 71% increase in pH stability, and +0.103 KJ/mole increase in ΔΔG, while the catalytic efficiency and ΔΔG value of L 287 R mutant increased by 4 times and +0.0903 KJ/mole, respectively. Thus, the mutants have the potential to be used in feed industries to increase the bioavailability of minerals while decreasing soil and water pollution.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0944-1344 1614-7499
DOI:	10.1007/s11356-022-18578-4