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Purification and Trypsin Inhibitor Activity of a Sporamin B from Sweet Potato (Ipomoea batatas Lam. 55-2)

文献类型： 外文期刊

作者： Sun Yan-li 1 ; Sun Jun-mao 2 ; Li Qing-peng 1 ;

作者机构： 1.Chinese Acad Agr Sci, Key Lab Agr Prod Proc & Qual Control, Minist Agr, Inst Agrofood Sci & Technol, Beijing 100193, Peoples R China

2.Chinese Acad Agr Sci, Agr Informat Inst, Beijing 100081, Peoples R China

关键词： sweet potato;sporamin B;identification;trypsin inhibitor activity

期刊名称：AGRICULTURAL SCIENCES IN CHINA （ 影响因子：0.82； 五年影响因子：0.997 ）

ISSN： 1671-2927

年卷期： 2009 年 8 卷 7 期

页码：

收录情况： SCI

摘要： Sporamin is a soluble protein in sweet potato, and falls into two distinct homology groups, subfamilies A and B. In this research, a sporamin B was purified and its amino acid sequences, trypsin inhibitor activity (Ti activity) were analyzed. This sporamin B was isolated from sweet potato tubers [Ipomoea batatas (L.) Lam cv. 55-2] through extraction of the water-soluble fraction, dialysis, ultrafiltration and ion-exchange chromatography. Homology determined by polyacrylamide gel electrophoresis showed that mainly one bond appeared in gel after being reduced by SDS (sodium dodecyl sulfate), or by SDS and 2-mercaptoethanol, or in native situation. By comparing the data of the polypeptide mass Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry with those of the mass of the theoretical amino acid sequences from NCBI protein database, it was revealed that it was Q40091|Q40091_IPOBA, sweet potato sporamin B Ipomoea Batatas (sweet potato) (Batate). The sequence coverage was 70.6%. N-terminal sequence was SETPV (Ser-Glu-Thr-Pro-Val). There is a linear relationship between trypsin inhibitor activity (Ti activity) and amounts of this sporamin B (3-18 mu g mL(-1)). The equation of linear regression was y = 2.5809x + 17.049 (r(2) = 0.9966). There was a curvilinear relationship between Ti activity and amounts of this sporamin B (21-150 mu g mL(-1)). The equation of curvilinear regression is y = 14.4171n(x)+ 23.26 (r(2) = 0.9924). The concentration of sporamin 13 with Ti activity after heating at 40 degrees C may induce part denature of this sporamin B, and there was no statistic difference after heating at 40, 50, 60 degrees C for 20 min. Heat treatment at more than 90 degrees C leads to a dramatic decrease of trypsin inhibitor efficiency. The results suggested that Q40091|Q40091_IPOBA was the major sporamin B in sweet potato tubers [Ipomoea Batatas (L.) Lam cv. 55-2], which had strong Ti activity, and was stable to both thermal and DTT (DL-dithiothreitol) relatively.

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