Biotechnology Reports (Mar 2022)

Isolation and characterization of a recombinant class C acid phosphatase from Sphingobium sp. RSMS strain

  • Shyam Sunder Rangu,
  • Rahul Singh,
  • Neeraj Kailash Gaur,
  • Devashish Rath,
  • Ravindra D. Makde,
  • Rita Mukhopadhyaya

Journal volume & issue
Vol. 33
p. e00709

Abstract

Read online

Tributyl phosphate (TBP) is extensively used in nuclear industry and is a major environmental pollutant. The mechanism for TBP degradation is not identified in any TBP-degrading bacteria. Here, we report identification of an acid phosphatase from Sphingobium sp. RSMS (Aps) that exhibits high specific activity towards monobutyl phosphate (MBP) and could be a terminal component of the TBP degradation process. A genomic DNA library of the bacteria was screened using a histochemical method which yielded 35 phosphatase clones. Among these, the clone that showed the highest MBP degradation was studied further. DNA sequence analysis showed that the genomic insert encodes a protein (Aps) which belongs to class C acid phosphatase. The recombinant Aps was found to be a dimer and hydrolysed MBP with a Kcat 68.1 ± 5.46 s−1 and Km 2.5 mM ± 0.50. The protein was found to be nonspecific for phosphatase activity and hydrolyzed disparate organophosphates.

Keywords