Cold-adapted protease derived from <i>Pseudoalteromonas arctica </i>PAMC 21717 and uses thereof

摘要

A cold-adapted protease derived from Pseudoalteromonas arctica PAMC 21717 or a recombinant cold-adapted protease obtained by expressing a gene encoding the cold-adapted protease in E. coli is described, and more particularly, a crystal of a protease exhibiting activity at low temperatures, a method for crystallizing the protease, a method for preparing the protease, a recombinant microorganism that expresses the protease, a method for preparing the recombinant microorganism, a method for preparing the recombinant protease and the use of the cold-adapted protease. The cold-adapted protease exhibits high activity at low temperatures, and securely maintains its enzymatic activity even in the presence of high pH, various metal ions and surfactants. Thus, it is useful in various industrial applications.

主权项

1. A crystal of a cold-adapted protease prepared by expressing a gene in a recombinant E. coli, said gene obtained by PCR using a template of DNA derived from Pseudoalteromonas arctica PAMC 21717 and a set of primers of SEQ ID NOs: 7 and 8, wherein the crystal of the cold-adapted protease has the following crystal structure:
(i) a subtilisin-like fold; ii) four calcium ions and two disulfide bonds (Cys439-Cys442 and Cys207-Cys254); (iii) a three-dimensional structure including ten α-helices which surround the central 6 β-strands and two β-strands; and (iv) a structure crystallized with P212121 space group either unit cell parameters of a=47.9 Å, b=74.6 Å, c=83.0 Å, α=β=γ=90° or 2374 atomic coordinates (including 4 calcium ions) set forth in Table 1.