Purification of antifreeze protein from wheat bran (Triticum aestivum L.) based on its hydrophilicity and ice-binding capacity.
Wheat-bran ( Triticum aestivum L.) antifreeze protein ( TaAFP) was purified 323-fold to electrophoretic homogeneity with an overall yield of 1.64% from wheat-bran protein by a specific three-step procedure. The three-step procedure was quicker, cheaper, and more effective than the five-step procedure we used earlier. First, TaAFP was concentrated by a phosphate buffer, on the basis of its strong hydrophilicity that was validated by thermal gravimetric analyses and a surface hydrophobicity analysis. Second, TaAFP was trapped in ice crystals for its specific ice-binding capacity, which was proved by ice-binding protocols. Remarkably, the ice-binding step was the most effective step, and the purification factor of this step was up to 270-fold. Finally, TaAFP was purified by HPLC purification, a complementary step for the specific ice-binding protocol, to electrophoretic homogeneity. Our protocols provide peers a novel and effective way for the search and purification of potential AFPs.
Zhang
Chao
C
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
Zhang
Hui
H
Wang
Li
L
Zhang
Junhui
J
Yao
Huiyuan
H
D021301:Antifreeze Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D004043:Dietary Fiber / Q000032:analysis*; D057927:Hydrophobic and Hydrophilic Interactions; D007053:Ice; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification
C114788:AFP protein, Daucus carota; D021301:Antifreeze Proteins; D004043:Dietary Fiber; D007053:Ice; D010940:Plant Proteins