3VM5
Recombinant medaka fish alpha-amylase expressed in yeast Pichia pastoris
Summary for 3VM5
| Entry DOI | 10.2210/pdb3vm5/pdb |
| Descriptor | alpha-amylase, CALCIUM ION, CHLORIDE ION, ... (4 entities in total) |
| Functional Keywords | (alpha/beta)8 barrel fold, starch hydrolysis, hydrolase |
| Biological source | Oryzias latipes |
| Total number of polymer chains | 1 |
| Total formula weight | 56786.38 |
| Authors | Mizutani, K.,Toyoda, M.,Mikami, B. (deposition date: 2011-12-08, release date: 2012-06-06, Last modification date: 2024-11-20) |
| Primary citation | 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 Cited by 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: 22613096DOI: 10.1016/j.bbapap.2012.05.005 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.85 Å) |
Structure validation
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