Uses
The enzyme has an optimal temperature of approximately 70 °C, but for a long incubation, a temperature below 60 °C is more suitable. It is stable in the pH range of 5 - 8.
General Description
Nuclease P1 is one of the most commonly known single-strand specific nucleases in molecular biology; Nuclease P1 is a single stranded specific endoduclease (of ssDNA or ssRNA). Nuclease P1 can also cleave single-stranded regions in double-stranded nucleic acids.
Nuclease P1 from
Penicillium citrinum is a zinc-dependent endonuclease that exhibits increased activity in the presence of low concentrations of urea. Nuclease P1 selective activity has found useful applications in studies on nucleic acid structure.
Biochem/physiol Actions
Catalyzes the nonspecific endonucleolytic cleavage of single stranded DNA and RNA to yield nucleoside 5′-phosphates and 5′-phosphooligonucleotides. It does not appreciably degrade double-stranded nucleic acids, especially in the presence of more than 400 mM sodium chloride at pH 6.0.
Synthesis
There are three main production methods for nuclease P1: solid-state fermentation, deep liquid fermentation and immobilized liquid fermentation. Solid-state fermentation (solid-state fermentation, SSF) operation is simple, low cost, the use of bran and other crops as raw materials, but also be able to get a higher enzyme activity of the crude enzyme liquid. However, the composition of these agricultural products is very complex, thus, it is more difficult and cumbersome to obtain pure nuclease P1.
The deep liquid fermentation method utilizes modern fermentation technology, and through various sensors installed in the fermenter, the fermentation process can be better controlled, and the ratio of its raw materials is also clearer. Usually the main conditions of the liquid fermentation process: synthetic liquid medium, ventilation 1:0.4, fermentation temperature 30 , incubation time of 50 ~ 70 h. Different manufacturers of the process conditions based on their own use of strains and equipment have to be adjusted. Overall, liquid fermentation of mycelium and fermentation broth is easier to separate, is currently the main method of production of nuclease P1 crude enzyme liquid at home and abroad, widely used in nucleotide fermentation.
