Simple method for isolation of glyceraldehyde 3-phosphate dehydrogenase and the improvement of myofibril gel properties.
Porcine glycolytic 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.
College of Agriculture, Ibaraki University, Ami, Ibaraki, Japan. miyaguti@mx.ibaraki.ac.jp
Yuji
Y
Miyaguchi
Taro
T
Sakamoto
Shun
S
Sasaki
Koji
K
Nakade
Manabu
M
Tanabe
Satoko
S
Ichinoseki
Masahiro
M
Numata
Kiichi
K
Kosai
D000645:Ammonium Sulfate; D000818:Animals; D002627:Chemistry, Physical; D004492:Edetic Acid; D004655:Emulsions; D005782:Gels; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification*; D008461:Meat Products; D008855:Microscopy, Electron, Scanning; D009210:Myofibrils / Q000648:ultrastructure; D011189:Potassium Chloride; D013552:Swine
D004655:Emulsions; D005782:Gels; D011189:Potassium Chloride; D004492:Edetic Acid; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000645:Ammonium Sulfate