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https://doi.org/10.31895/hcptbn.15.1-2.6

Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity

Sunčica Beluhan ; Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia
Ivana Karmelić ; School of Medicine, University of Zagreb, Zagreb, Croatia
Mirela Ivančić Šantek ; Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Puni tekst: engleski, pdf (960 KB) str. 27-37 preuzimanja: 46* citiraj
APA 6th Edition
Beluhan, S., Karmelić, I. i Ivančić Šantek, M. (2020). Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity. Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam, 15 (1-2), 27-37. https://doi.org/10.31895/hcptbn.15.1-2.6
MLA 8th Edition
Beluhan, Sunčica, et al. "Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity." Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam, vol. 15, br. 1-2, 2020, str. 27-37. https://doi.org/10.31895/hcptbn.15.1-2.6. Citirano 24.01.2021.
Chicago 17th Edition
Beluhan, Sunčica, Ivana Karmelić i Mirela Ivančić Šantek. "Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity." Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam 15, br. 1-2 (2020): 27-37. https://doi.org/10.31895/hcptbn.15.1-2.6
Harvard
Beluhan, S., Karmelić, I., i Ivančić Šantek, M. (2020). 'Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity', Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam, 15(1-2), str. 27-37. https://doi.org/10.31895/hcptbn.15.1-2.6
Vancouver
Beluhan S, Karmelić I, Ivančić Šantek M. Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity. Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam [Internet]. 2020 [pristupljeno 24.01.2021.];15(1-2):27-37. https://doi.org/10.31895/hcptbn.15.1-2.6
IEEE
S. Beluhan, I. Karmelić i M. Ivančić Šantek, "Adding value in barley malt rootlets as a source of 5’-phosphodiesterase:biochemical and thermodynamic evaluation of enzyme activity", Hrvatski časopis za prehrambenu tehnologiju, biotehnologiju i nutricionizam, vol.15, br. 1-2, str. 27-37, 2020. [Online]. https://doi.org/10.31895/hcptbn.15.1-2.6

Sažetak
thermostable 5’-phosphodiesterase (5’-PDE, EC 3.1.4.1) was extracted from barley (Hordeum distichum var. Rex) malt rootlets. The purification procedure comprised acetone precipitation, S-Sepharose cation-exchange and DEAE-Sepharose anion-exchange chromatography. The enzyme was purified 101-fold with a recovery of 22% and a specific activity of 81.9 U mg-1 protein, Optimum enzyme activity was obtained at 70 °C, and pH 8.9. The SDS-PAGE profiling of the purified protein exhibited molecular weight of 116 kDa and revealed three sub-unit fractions of 26, 43, and 56 kDa making up its active configuration. The kinetic constants Km and Vmax were determined as 0.25 mM and 0.816 mmol min-1, respectively. Thermodynamic studies showed that the thermal inactivation of purified barley malt rootlets 5’-PDE followed the first-order kinetics, indicating inactivation energy (Ed) of 134 kJ mol-1. The half-life (t1/2) at 70 °C was estimated as 169 min. Thermodynamic parameters ΔH*, ΔS* and ΔG* were determined as a function of temperature and were 131.15 kJ mol-1, 37.01 kJ mol-1 K-1 and 118.4 kJ mol-1, respectively. The purified enzyme has long half-life with 11 days at 0 °C, 37 hours at 4 °C and 11 hours at room temperature. These results provide useful information about the factors that affects the activity of barley malt rootlets 5’-PDE and suggests a good indication for application of this enzyme in pharmaceutical and food industry.

Ključne riječi
barley malt rootlets, 5’-phosphodiesterase, pH stability, thermal stability, thermodynamic parameters

Hrčak ID: 245698

URI
https://hrcak.srce.hr/245698

Posjeta: 94 *