A One-Step Protocol for Chromatographic Purification

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for routine bacterial growth at 37 °C. Fractional salt precipitation was carried out using ammonium sulfate. The salt was added to the bacterial culture to reach 40 ...
BioNanoSci. DOI 10.1007/s12668-016-0226-9

A One-Step Protocol for Chromatographic Purification of Non-recombinant Exogenous Bacterial Enzyme: Nuclease of Serratia marcescens Gulnaz Vafina 1 & Emil Bulatov 1

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Elmira Zaynutdinova 1 & Maria Filimonova 1

# Springer Science+Business Media New York 2016

Abstract Serratia marcescens are well-known Gram-negative bacteria capable of excreting extracellular hydrolases, such as the nuclease—an enzyme that catalyzes hydrolysis of both DNA and RNA chains with a high efficiency. The nuclease has a number of attractive properties for use in biotechnology industry—outstanding enzymatic activity and low production cost. The existing protocols for purification of nuclease yield only limited amounts of protein and require complicated multistage procedures. Here, we report a chromatographic protocol for elegant one-step purification procedure resulting in a pure homogenous enzyme, as confirmed by gel electrophoresis and mass spectrometry. Keywords Serratia marcescens nuclease . Chromatographic purification . Homogeneous enzyme . Gel electrophoresis . Mass spectrometry

1 Introduction Serratia marcescens are Gram-negative bacteria of the Enterobacteriaceae family that can lead to a number of infective diseases in humans. However, despite the pathogenic nature of S. marcescens, the nuclease expressed by this organism has a number of uses in biotechnology industry and is currently marketed under trademark Benzonase®. The nuclease non-specifically cleaves all types of nucleic acids into small oligonucleotides (