Keywords

Polyacrylic Acid, Alkaline Protein, Sds-Page, Ief, Bitter Melon

Abstract

Using balsam pear seeds as material, the method and influencing factors of polyacrylic acid separation and purification of basic protein from balsam pear kernels were studied. The isoelectric point precipitation test showed that when citric acid and hydrochloric acid adjusted the pH of the crude extract of bitter melon seeds to 6.0 and 4.0, respectively, 14.62% and 32.49% of the bitter melon seed kernel proteins were precipitated. The isoelectric point precipitation of acetic acid shows stage characteristics. At pH 6.0 and 4.0, 26.17% and 38.72% of the bitter melon kernel protein are precipitated respectively. For 1 ml crude extract of bitter melon seeds (ph 4.0) treated with acetic acid, hydrochloric acid and citric acid, the optimal dosage of 1% paa for selective precipitation of basic protein (isoelectric point pi is 8.65-9.30) is 100 ¦Ìl, 120 ¦Ìl and 100¦Ìl. Acetic acid adjusts the pH of the crude extract of bitter melon seeds to 5.0, 4.0 and 3.0 respectively. The supernatant after isoelectric point precipitation is used to precipitate alkaline protein with paa. When the dosage of paa (1%) is 160¦Ìl/ml of the extract, the pH is 5. 33.77% and 43.56% of the protein were precipitated in the 0 and 3.0 sample solutions respectively; when the paa dosage was 120 ¦Ìl/ml extraction solution, 30.83% of the protein in the ph4.0 sample solution was precipitated. The paa-protein complex is dissolved in alkaline solution (ph>9.0). When the NaCl concentration of the solution is 3.0%, the protein concentration of the solution is the highest. The basic protein from balsam pear kernels selectively precipitated by paa was separated by sephadexg-75 column chromatography, and the main peaks ¢¡ and ¢¢ appeared at 175 min and 300 min respectively. SDS-PAGE and IEF analysis show that the molecular weight of main peak ¢¡ is about 30kd, and the pi value is about 9.5. The molecular weight of main peak ¢¢ is about 10kd, and the pi value is about 9.3.

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Original research done by Zhang Haihua, Wang Qiaomei, Hu Jiasu, Tong Fudan

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