Purification of L-Glutaminase from Bacillus sp. B12 and study its properties

Authors

  • Alyaa Abdelhameed1, Rana Alqaysi1, Hiba Ali Hilal2 , and Nada Alharbi 3

DOI:

https://doi.org/10.37506/ijfmt.v14i2.3063

Keywords:

Bacillus sp.B12, L- glutaminase, purification, Ion exchange , pH, and Thermostability

Abstract

L-glutaminase is an enzyme that catalyse the conversion L-glutamine to glutamic acid and ammonia. The important application of the L-glutaminase is a chemotherapy agent. In this study a novel strain, Bacillus B12. that isolated from Diyala soil was explored for production of extracellular L-glutaminase. The enzyme has been purified 23.9-fold from cell-free extract with 45.4 recovery ( specific activity 76.6 U/mg protein). Enzyme has molecular weight of 199 kDa, and pH 8 and stable in pH range 6-9.5. Temperature optimum is 40 ?C and completely stable between 25-45 ?C. The kinetics studies revealed that the km and Vmax of purified L-glutaminase were estimated in 0.4mmol/L and 0.133mmol/min, respectively. The result showed that, the enzyme was active when incubated with 10 mM of Mn2+, Mg2+,Ca+2 and Na+. Whereas, K +, Co2+, and Ni2+ show no effect. However, Hg2+, Cu2+,Fe2+ and Zn2+ decreased L-glutaminase activity . The 10mM of DEAE, PMSF, and sodium azide did not show a clear effect against the enzyme activity. Higher decrease in enzyme activity was observed by using Cysteine and 2-Mercaptoethanol ( 30 and 25%, respectively).

Author Biography

Alyaa Abdelhameed1, Rana Alqaysi1, Hiba Ali Hilal2 , and Nada Alharbi 3

1Biotechnology department -College of Science-University of Diyala–Iraq, 2Microbiology department- College of Veterinary medicine-– University of Diyala–Iraq, 3Biology department, College of Science- University of Princess Nourah bint Abdulrahman -Saudi Arabia

Published

2020-04-29

How to Cite

Alyaa Abdelhameed1, Rana Alqaysi1, Hiba Ali Hilal2 , and Nada Alharbi 3. (2020). Purification of L-Glutaminase from Bacillus sp. B12 and study its properties. Indian Journal of Forensic Medicine & Toxicology, 14(2), 1160-1166. https://doi.org/10.37506/ijfmt.v14i2.3063