Premium
Simple method for isolation of glyceraldehyde 3‐phosphate dehydrogenase and the improvement of myofibril gel properties
Author(s) -
MIYAGUCHI Yuji,
SAKAMOTO Taro,
SASAKI Shun,
NAKADE Koji,
TANABE Manabu,
ICHINOSEKI Satoko,
NUMATA Masahiro,
KOSAI Kiichi
Publication year - 2011
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1111/j.1740-0929.2010.00807.x
Subject(s) - myofibril , chemistry , chromatography , dehydrogenase , glyceraldehyde 3 phosphate dehydrogenase , affinity chromatography , salting out , sepharose , phosphate , nuclear chemistry , biochemistry , enzyme , aqueous solution , organic chemistry
Porcine glycoliytic enzyme, glyceraldehyde 3‐phosphate dehydrogenase (G3PD) was prepared effectively by a combination of ethylene diamine tetra‐acetate (EDTA) pretreatment and affinity purification. After salting out of porcine sarcoplasmic proteins (SP) with ammonium sulfate at 75% saturation, the obtained supernatant (SP‐f3) was treated with EDTA, leaving G3PD in the supernatant (G3PD‐E) and most other SPs in the precipitate. At that time, the separation of G3PD‐E required more than 20 mmol/L EDTA. G3PD‐E was then subjected to affinity purification by batchwise method using blue‐sepharose CL‐6B, and purified G3PD (G3PD‐AP) was obtained using 2 mol/L potassium chloride (KCl) as an eluent. Texture analysis showed that the hardness, adhesiveness and gumminess of the myofibril gel at 0.2‐mol/L NaCl increased with the addition of G3PD‐AP. Scanning electron microscopy revealed that the G3PD‐AP reinforced the gel network of the myofibril. However, scanning electron micrograph analysis showed that the network‐structure of the gel by the addition of G3PD‐AP developed in a different manner from that by adding 0.6 mol/L NaCl. These results showed that glycolytic enzyme, G3PD, contributes to the improvement of the rheological properties of meat products.