Highly efficient novel recombinant L-asparaginase with no glutaminase activity from a new halo-thermotolerant Bacillus strain

BioImpacts : BI
Azam SafarySiavoush Dastmalchi

Abstract

Introduction: The bacterial enzyme has gained more attention in therapeutic application because of the higher substrate specificity and longer half-life. L-asparaginase is an important enzyme with known antineoplastic effect against acute lymphoblastic leukemia (ALL). Methods: Novel L-asparaginase genes were identified from a locally isolated halo-thermotolerant Bacillus strain and the recombinant enzymes were overexpressed in modified E. coli strains, OrigamiTM B and BL21. In addition, the biochemical properties of the purified enzymes were characterized, and the enzyme activity was evaluated at different temperatures, pH, and substrate concentrations. Results: The concentration of pure soluble enzyme obtained from Origami strain was ~30 mg/L of bacterial culture, which indicates the significant improvement compared to L-asparaginase produced by E. coli BL21 strain. The catalytic activity assay on the identified L-asparaginases (ansA1 and ansA3 genes) from Bacillus sp. SL-1 demonstrated that only ansA1 gene codes an active and stable homologue (ASPase A1) with high substrate affinity toward L-asparagine. The Kcat and Km values for the purified ASPase A1 enzyme were 23.96s-1 and 10.66 µM, respectively. In addition, the rec...Continue Reading

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Citations

Dec 7, 2019·BMC Research Notes·Raquel Caminha DantasKaio Cesar Simiano Tavares
Jan 29, 2020·BioImpacts : BI·Marziyeh FathiJaleh Barar
May 19, 2019·Cellular and Molecular Life Sciences : CMLS·Azam SafaryMohammad A Rafi
May 19, 2020·Biomedit︠s︡inskai︠a︡ khimii︠a︡·M V DuminaN N Sokolov
Sep 12, 2019·BioImpacts : BI·Mostafa Akbarzadeh KhiaviMohammad Hossein Somi
Jun 2, 2021·Applied Microbiology and Biotechnology·Daniel CastroValéria C Santos-Ebinuma
Sep 18, 2021·Environmental Science and Pollution Research International·Sulaiman A Al Yousef

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Datasets Mentioned

BETA
KX129701

Methods Mentioned

BETA
protein assay
PCR

Software Mentioned

Jalview
ProtParam
BLASTn
ExPASy
ClustalW
Prism
GraphPad

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