New Nuclease from Extremely Psychrophilic Microorganism Psychromonas ingrahamii 37: Identification and Characterization

• Published in: Molecular biotechnology 2019-02, Vol.61 (2), p.122-133
• Main Authors: Maciejewska, Natalia, Walkusz, Rafał, Olszewski, Marcin, Szymańska, Aneta
• Format: Article
• Language: English
• Subjects: Biochemistry; Biological Techniques; Biotechnology; Cell Biology; Chemistry; Chemistry and Materials Science; Human Genetics; Original Paper; Protein Science
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-018-0142-z

Nucleases are an important group of hydrolases that degrade nucleic acids, with broad spectrum of applications in science and industry. In this paper, we report the identification and characterization of the nuclease from extremely psychrophilic bacterium Psychromonas ingrahamii that grows exponentially at 5 °C, but may also grow at even lower temperatures (down to − 12 °C). The putative endonuclease I gene, identified in the genome of P. ingrahamii , was cloned and expressed in Pichia pastoris . The recombinant protein was purified and its nucleolytic features were studied. The new enzyme, named by us as Pin Nuc, displays the features characteristic for the nonselective endonucleases, and has the ability to degrade different forms of nucleic acids. It is very active at room temperature in low ion-strength buffer and in the presence of low concentrations of magnesium ions. The enzyme, which possesses six cysteine residues, the most likely all engaged in disulphide bridges, is active only in oxidized form, and can be efficiently inactivated by the addition of low amounts of a reducing agent. According to our knowledge, it is the first nuclease, belonging to endonuclease I family, isolated from such extremely psychrophilic organism.