Unknown

Dataset Information

0

A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application.


ABSTRACT: L-asparaginase (L-ASNase) is a vital enzyme with a broad range of applications in medicine and food industry. Drawbacks of current commercial L-ASNases stimulate the search for better-producing sources of the enzyme, and extremophiles are especially attractive in this view. In this study, a novel L-asparaginase originating from the hyperthermophilic archaeon Thermococcus sibiricus (TsA) was expressed in Escherichia coli, purified and characterized. The enzyme is optimally active at 90 °C and pH 9.0 with a specific activity of 2164 U/mg towards L-asparagine. Kinetic parameters KM and Vmax for the enzyme are 2.8 mM and 1200 µM/min, respectively. TsA is stable in urea solutions 0-6 M and displays no significant changes of the activity in the presence of metal ions Ni2+, Cu2+, Mg2+, Zn2+ and Ca2+ and EDTA added in concentrations 1 and 10 mmol/L except for Fe3+. The enzyme retains 86% of its initial activity after 20 min incubation at 90 °C, which should be enough to reduce acrylamide formation in foods processed at elevated temperatures. TsA displays strong cytotoxic activity toward cancer cell lines K562, A549 and Sk-Br-3, while normal human fibroblasts WI-38 are almost unsensitive to it. The enzyme seems to be a promising candidate for further investigation and biotechnology application.

SUBMITTER: Dumina M 

PROVIDER: S-EPMC8470970 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC92756 | biostudies-literature
| S-EPMC8146568 | biostudies-literature
| S-EPMC7657641 | biostudies-literature
| S-EPMC4959179 | biostudies-literature
| S-EPMC2432063 | biostudies-literature
| S-EPMC2685551 | biostudies-literature
| S-EPMC148058 | biostudies-literature
| S-EPMC4247193 | biostudies-literature
| S-EPMC3993132 | biostudies-literature
| S-EPMC4551206 | biostudies-literature