High-yield novel leech hyaluronidase to expedite the preparation of specific hyaluronan oligomers.

ABSTRACT: Hyaluronidases (HAases), particularly leech HAases, have attracted intense attention due to their broad applications in medical treatments and great potential for the enzymatic production of hyaluronan oligosaccharides. However, little is known about this third interesting family of HAases. Here, we applied the random amplification of cDNA ends polymerase chain reaction (RACE-PCR) approach to identify the first leech HAase-encoding gene. By combining protein engineering and high-density culture, we achieved high-level production (8.42 × 10(5) U ml(-1)) in the yeast Pichia pastoris secretory expression system. Compared with the commercial bovine testicular HAase, the recombinant leech HAase exhibited superior enzymatic properties. Furthermore, analysis of the hydrolytic process suggested th

SUBMITTER: Jin P 

PROVIDER: S-EPMC3966032 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature