3VM5

Recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris

3VM5 の概要

• エントリーDOI: 10.2210/pdb3vm5/pdb
• 分子名称: alpha-amylase, CALCIUM ION, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: (alpha/beta)8 barrel fold, starch hydrolysis, hydrolase
• 由来する生物種: Oryzias latipes
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 56786.38

構造登録者

Mizutani, K.,Toyoda, M.,Mikami, B. (登録日: 2011-12-08, 公開日: 2012-06-06, 最終更新日: 2024-11-20)

主引用文献

Mizutani, K.,Toyoda, M.,Otake, Y.,Yoshioka, S.,Takahashi, N.,Mikami, B.
Structural and functional characterization of recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris.
Biochim.Biophys.Acta, 1824:954-962, 2012

PubMed Abstract: The medaka fish α-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 α-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 (α/β)(8) barrel fold, as do other known α-amylases, and the overall structure is very similar to the structure of vertebrate (human and pig) α-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.

PubMed: 22613096
DOI: 10.1016/j.bbapap.2012.05.005

実験手法

X-RAY DIFFRACTION (2.85 Å)