Affiliation:
1. Nanjing Tech University
Abstract
Abstract
L-asparaginase (ASNase), as a pivotal amidohydrolase enzyme, has been used in removing acrylamide in food processing and treating acute lymphoblastic leukaemia in clinic. In this study, a novel ASNase from Bifidobacterium thermophilum (BtASNase) was successfully cloned and heterologously expressed in E. coli host. BtASNase was identified to share maximum 40% structural similarity with other ASNases in PDB database. The purified BtASNase with monomeric size about 35 kDa had the highest specific activity (554.82 IU/mg) at 55℃ and pH 8.0. Further investigation indicated that BtASNase showed great stability at wide pH range (6.0–11.0), and retained more than 85% of its activity for 50 min at 37℃. To be noted, BtASNase exhibited high L-asparaginase specificity and zero glutaminase activity. To our knowledge, this is the first time to explore ASNase from Bifidobacterium thermophilum, and explored BtASNase could be a potential candidate with desirable advantages for unraveling glutaminase activity, narrow pH range stability, and low thermostability restrict in industry applications of ASNase.
Publisher
Research Square Platform LLC
Reference35 articles.
1. In vitro and in vivo killing of acute lymphoblastic leukemia cells by L-asparaginase;Asselin BL;Cancer Res,1989
2. L-asparaginase as a critical component to combat Acute Lymphoblastic Leukaemia (ALL): A novel approach to target ALL;Ali U;Eur J Pharmacol,2016
3. )Bacillus sonorensis L.Asparaginase: Cloning,expression in E.coli and characterization;Aly NH;The Protein Journal,2020
4. Alrumman SA, Mostafa YS, Al-izran KA, Alfaifi MY, Taha T, Elbehairi SEI (2019) Production and anticancer activity of an L-Asparaginase from Bacillus licheniformis isolated from the Red Sea, Saudi Arabia. Sci Rep 9:1–14. Doi:10.1038/s41598-019-40512-x.
5. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding;Bradford MM;Anal Biochem,1976