In silico analysis, molecular cloning, expression and characterization of l-asparaginase gene from Lactobacillus reuteri DSM 20016.

ABSTRACT: l-Asparaginase is employed in leukaemic treatment and in processing starchy foods. The in silico analysis of Lactobacillus reuteri DSM 20016 reveals the presence of an l-asparaginase gene with theoretical pI value of 4.99. 3D structure prediction was carried out and one model was selected based on the validation scores of 86.293 for ERRAT, 92.10% for VERIFY 3D and Ramachandran plot. Multiple sequence alignment of the protein sequences of l-asparaginases I and II of Escherichia coli, Erwinia chrysanthemum and Homo sapiens shows their sequence similarity. The ORF LREU_RS09880 from L. reuteri DSM 20016 genome was cloned and expressed in E. coli. The recombinant protein was purified to homogeneity using Ni-NTA chromatography and showed higher substrate

SUBMITTER: Susan Aishwarya S 

PROVIDER: S-EPMC5612876 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature