PMID: 45489May 1, 1977

Isolation and characterization of a polynucleotide phosphorylase from Bacillus amyloliquefaciens

Journal of Bacteriology
R J Erickson, J C Grosch

Abstract

Bacillus amyloliquefaciens BaM-2 produces large amounts of extracellular enzymes, and the synthesis of these proteins appears to be dependent upon abnormal ribonucleic acid metabolism. A polynucleotide phosphorylase (nucleoside diphosphate:polynucleotide nucleotidyl transferase) was identified, purified, and characterized from this strain. The purification scheme involved cell disruption, phase partitioning, differential (NH4)2SO4 solubilities, agarose gel filtration, and diethylaminoethyl-Sephadex chromatography. The purified enzyme demonstrated the reactions characteristic of polynucleotide phosphorylase: polymerization, phosphorolysis, and inorganic phosphate exchange with the beta-phosphate of a nucleotide diphosphate. The enzyme was apparently primer independent and required a divalent cation. The reactions for the synthesis of the homopolyribonucleotides, (A)n and (G)n, were optimized with respect to pH and divalent cation concentration. The enzyme is sensitive to inhibition by phosphate ion and heparin and is partially inhibited by rifamycin SV and synthetic polynucleotides.

Related Concepts

PNPT1
Transcriptase
Rifamicina Colirio
Rifamycins
Poly(A) Tail
Ion-Exchange Chromatography Procedure
Molecular Sieve Chromatography
Polyribonucleotide Nucleotidyltransferase
Hydrogen-Ion Concentration
Magnesium ADP

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