Heterologous Expression and Enzymatic Characterization of Adenylyl-Sulfate Kinase from Mycobacterium tuberculosis

Abstract:

PAPS is the only active sulfonic acid group donor for sulfation in living organisms. Adenylyl-sulfate kinase catalyzes the formation of PAPS and ADP from APS and ATP. In order to achieve the enzymatic synthesis of PAPS, the adenylyl-sulfate kinase encoding gene from Mycobacterium tuberculosis AUSMDU00018547 was codon-optimized and heterologously co-expressed with thioredoxin TrxB to achieve the active and high-level heterologous expression of adenosine phosphorylsulfate kinase (APK) in Escherichia coli BL21(DE3). His-tagged recombinant adenylyl-sulfate kinase was successfully purified. Further enzymatic property analysis results showed that the optimal temperature, pH and the specific enzyme activity of recombinant adenylyl-sulfate kinase were 35 ℃, pH 7.5 and (1.26±0.08) U/mg, respectively. Heterologous expression of adenylyl-sulfate kinase and enzymatic property analysis lay a foundation for the enzymatic synthesis of PAPS in the future.