Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

A Novel Type II L-Asparaginase from the Halotolerant Bacillus subtilis CH11

Annsy Arredondo-Nuñez
ORCID logo ,
Gisele Monteiro
ORCID logo ,
Carol Flores-Fernández
* ,
Lina Antenucci
,
Perttu Permi
,
Amparo Iris Zavaleta
*
Version 1 : Received: 29 September 2023 / Approved: 30 September 2023 / Online: 30 September 2023 (08:39:19 CEST)

A peer-reviewed article of this Preprint also exists.

Arredondo-Nuñez, A.; Monteiro, G.; Flores-Fernández, C.N.; Antenucci, L.; Permi, P.; Zavaleta, A.I. Characterization of a Type II L-Asparaginase from the Halotolerant Bacillus subtilis CH11. Life 2023, 13, 2145. Arredondo-Nuñez, A.; Monteiro, G.; Flores-Fernández, C.N.; Antenucci, L.; Permi, P.; Zavaleta, A.I. Characterization of a Type II L-Asparaginase from the Halotolerant Bacillus subtilis CH11. Life 2023, 13, 2145.

Abstract

L-asparaginase from bacterial sources have been used in antineoplastic treatments and the food industry. A novel type II L-a sparaginase encoded by the N-truncated gene ansZP21 of halotolerant Bacillus subtilis CH11 isolated from Chilca salterns in Peru was expressed using a heterologous system in Escherichia coli BL21 (DE3)pLysS. The recombinant protein was purified using one-step nickel affinity chromatography and exhibited activity of 234.38 U mg-1 and a maximum catalytic activity at pH 9.0 and 60 °C. The enzyme shows a homotetrameric form with an estimated molecular weight of 155 kDa by gel filtration chromatography. The enzyme half-life at 60 °C was 3 h 48 min, and L-asparaginase retained 50% of initial activity for 24 h at 37 °C. The activity was considerably enhanced by KCl, CaCl2, MgCl2, mercaptoethanol, and DL-dithiothreitol (p-value < 0.01). Moreover, the Vmax and Km were 145.2 µmol mL-1 min-1 and 4.752 mM, respectively. These findings evidence a promising novel type II L-asparaginase for future industrial applications.

Keywords

L-asparaginase II; Bacillus subtilis; saline environment; biochemical characterization

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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