3vm5

Publication Abstract from PubMed

The medaka fish alpha-amylase was expressed and purified. The expression systems were constructed using methylotrophic yeast Pichia pastoris, and the recombinant proteins were secreted into the culture medium. Purified recombinant alpha-amylase exhibited starch hydrolysis activity. The optimal pH, denaturation temperature, and K(M) and V(max) values were determined; chloride ions were essential for enzyme activity. The purified protein was also crystallized and examined by X-ray crystallography. The structure has the (alpha/beta)(8) barrel fold, as do other known alpha-amylases, and the overall structure is very similar to the structure of vertebrate (human and pig) alpha-amylases. A novel expression plasmid was developed. Using this plasmid, high-throughput construction of an expression system by homologous recombination in P. pastoris cells, previously reported for membrane proteins, was successfully applied to the secretory protein.

Structural and functional characterization of recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris.,Mizutani K, Toyoda M, Otake Y, Yoshioka S, Takahashi N, Mikami B Biochim Biophys Acta. 2012 May 18. PMID:22613096^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.