鹰嘴豆分离蛋白的酶解工艺研究%Study on the Enzyme Hydrolysis of Chickpea Protein
河南省科学院生物研究所有限责任公司,郑州450008; 河南省微生物工程重点试验室,郑州450008 河南省科学院生物研究所有限责任公司,郑州,450008
2013-11-22
鹰嘴豆分离蛋白 短肽 碱性蛋白酶 水解度
研究鹰嘴豆分离蛋白的酶解工艺,提高鹰嘴豆蛋白水解度,为鹰嘴豆酶解生产短肽的产业化提供依据。用pH-state法计算蛋白水解度,首先通过单因素试验和正交试验,得出碱性蛋白酶水解鹰嘴豆蛋白的最佳反应条件,然后结合木瓜蛋白酶和菠萝蛋白酶,共同水解鹰嘴豆蛋白,以提高蛋白水解度。碱性蛋白酶水解鹰嘴豆分离蛋白的最佳反应条件是:pH 8.5、反应温度55℃、底物浓度[S]2%、酶与底物浓度之比([E]/[S])2%,在此条件下,蛋白水解度可以达到27.86%,碱性蛋白酶、木瓜蛋白酶、菠萝蛋白酶在各自最佳反应条件下(碱性蛋白酶pH 8.5、反应温度55℃、[S]2%、([E]/[S])2%,木瓜蛋白酶和菠萝蛋白酶均为pH 7.2、反应温度55℃、[S]2%、([E]/[S])2%)依次水解鹰嘴豆分离蛋白,反应结束时,蛋白水解度可达到34.64%;3种酶在各自最佳反应条件下依次水解鹰嘴豆分离蛋白,反应结束时,蛋白水解度显著高于三者同时作用,小分子短肽得率明显提高。%To increase the DH of chickpea protein, and provide the basis for industrialization of the enzymatic production of chickpea producing oligopeptides, the optimal condition for alcalase hydrolysis of chickpea protein was studied. The degree of hydrolysis (DH) was determined according to the pH-state method. Through a single-factor test and an orthogonal designed experiment, the alcalase hydrolysis of chickpea protein was systematically studied. Then, the three proteases of alcalase,papain and bromelain commonly hydrolyzing chickpea protein was carried out to increase the DH of chickpea protein. The results showed that the obtained optimum hydrolysis with alcalase condition was pH 8.5, temperature (T) 55℃, concentration of substrate [S] 2%, and ratio of enzyme and substate ([E]/[S]) 2%. Under this condition, the degree of hydrolysis (DH) was 27.86%. Under each optimum conditions of the three proteases respectively (The obtained optimum hydrolysis condition of alcalase was pH 8.5, temperature (T) 55℃, concentration of substrate [S] 2%, and ratio of enzyme and substate ([E]/[S]) 2%. The obtained optimum hydrolysis condition of papain and bromelain was pH 7.2, temperature (T) 55℃, concentration of substrate [S] 2%, and ratio of enzyme and substate ([E]/[S]) 2%), the chickpea protein hydrolyzing by the alcalase, papain and bromelain in turn was carried out, and the DH could reach 34.64%. The DH of chickpea protein hydrolyzing by the three proteases above-mentioned under each optimum condition respectively in turn was much higher than that of the DH of chickpea protein hydrolyzing by the three proteases simultaneously. Furthermore, the yield of oligopeptides was significantly improved.
