New nuclease from extremely psychrophilic microorganism Psychromonas ingrahamii 37: identification and characterization

Natalia Maciejewska , Rafał Walkusz , Marcin Olszewski , Aneta Szymańska

Abstract

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 PinNuc, 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.
Author Natalia Maciejewska (FCh / DBCh / LMCh)
Natalia Maciejewska,,
- Laboratory of Medical Chemistry
, Rafał Walkusz
Rafał Walkusz,,
-
, Marcin Olszewski
Marcin Olszewski,,
-
, Aneta Szymańska (FCh / DBCh / LMCh)
Aneta Szymańska,,
- Laboratory of Medical Chemistry
Journal seriesMolecular Biotechnology, ISSN 1073-6085, (A 25 pkt)
Issue year2019
Vol61
No2
Pages122-133
Publication size in sheets0.55
Keywords in EnglishNuclease, psychrophilic, substrate specificity, Psychromonas ingrahamii 37, enzymatic activity, cold-active enzyme
ASJC Classification1312 Molecular Biology; 2402 Applied Microbiology and Biotechnology; 1303 Biochemistry; 1502 Bioengineering; 1305 Biotechnology
DOIDOI:10.1007/s12033-018-0142-z
URL https://doi.org/10.1007/s12033-018-0142-z
Languageen angielski
LicenseOther; published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)25
ScoreMinisterial score = 20.0, 16-07-2019, ArticleFromJournal
Ministerial score (2013-2016) = 25.0, 16-07-2019, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2017 = 0.625; WoS Impact Factor: 2017 = 1.815 (2) - 2017=1.763 (5)
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