螺旋藻肽的制备及其抗氧化性研究

发布时间：2019-06-11 04:38

【摘要】：螺旋藻作为一种食品原料因其功能特性而得到广泛应用,FAO将其评为“人类21世纪最佳保健品”和“未来超级营养食品”(张文等,2013)。螺旋藻中藻胆蛋白含量高达50-70%。此外,还具有非常理想的氨基酸构成比例,人体所必需的8种氨基酸的含量接近或超过FAO规定的标准。螺旋藻对高胆固醇血症、高甘油血症、心血管疾病、炎症性疾病、癌症以及病毒性感染等一系列疾病有疗效。螺旋藻在食品制造业中还可以作为一种功能成分存在(Hemamalini,2013)。但是,螺旋藻在食品制造业中的应用受到其干粉溶解性差、腥味浓重的缺点的限制。藻胆蛋白经蛋白酶处理后,改善了物化特性并扩大了应用范围。与藻胆蛋白相比,其水解产物螺旋藻肽不仅具有较低的分子量与良好的物化特性,更容易被人体消化吸收。而且还具有提高人体免疫功能,消除并防止机体受到自由基破坏,全面调节人体代谢机能等生理作用,这其中以其抗氧化能力最受关注。本文主要研究了螺旋藻的破壁方法、优化了单酶法和双酶法水解螺旋藻制备抗氧化肽的工艺,研究了水解度对螺旋藻肽起泡性、乳化性及抗氧化能力的影响,还研究了酶解过程中色素的产生规律与防控措施以及螺旋藻肽功能性饮料的研制。搅拌法、超声波破碎法、反复冻融法及后二者联用都可以破坏螺旋藻藻体细胞,使内容物流出。研究结果表明,超声波破碎法,在超声模式相同、超声功率为5%、超声7s、间隔3s时,细胞中可溶性蛋白的含量是最多的;反复冻融法4℃下解冻的样品比37℃下解冻的样品溶液中可溶性蛋白的含量要高,冻融次数与可溶性蛋白含量之间存在正相关。两种方法联用时的破壁效果要好于单独使用一种方法。采用搅拌法破壁时,溶液中可溶性蛋白含量随着搅拌的进行含量增加,但是增速逐渐减缓,并且在搅拌时间为3h时,得到蛋白的量为25.8mg/m L。采用Box-Behnken法对碱性蛋白酶制备螺旋藻抗氧化肽的工艺条件进行优化,优化过程以DPPH自由基清除能力与收率为响应值。结果表明,相较于木瓜蛋白酶、胰蛋白酶与风味蛋白酶,碱性蛋白酶对螺旋藻的水解能力最强,其最适水解条件为温度55.46℃、pH 6.71、固液比为1:10.83(v/w)。在此条件下,酶解240min,螺旋藻肽的得率为58.50%,与优化前相比,提高了15.61%。经此法可获得具有明显抗氧化活性的螺旋藻肽,浓度为0.86g/L时,其DPPH自由基清除能力为77.60%,与优化前相比,提高了7.62%。采用中心组合实验对双酶水解螺旋藻制备螺旋藻抗氧化肽的生产工艺进行优化,响应值为DPPH自由基清除能力及收率。研究结果表明,双酶法的较佳工艺为温度54.89℃,碱性蛋白酶与木瓜蛋白酶的比为2.23:1,酶解时间6.53h,此时,理论上DPPH自由基清除能力与收率的最大值分别为78.00%和57.94%。水解度对螺旋藻肽的起泡性、乳化性及抗氧化能力有影响。研究结果表明,螺旋藻肽的水解度随着水解的进行逐渐增大。水解过程中,螺旋藻肽的起泡性、DPPH自由基清除能力、以及还原力均随着水解度的增加先增大后减小,羟基自由基清除能力随着水解的进行先减小后增大,达到最大后再减小。而泡沫稳定性、乳化性、乳化稳定性则随着水解度的增大逐渐减小。酶解过程中的色素随着水解的进行逐渐积累,适当添加抑制剂或者吸附剂可以有效的抑制色素的产生。水解开始时加入1.5%的H2O2,水解结束时色素抑制率可达31.07%;NaClO加入量为1.5%时,色素抑制率为21.47%;当加入3%的活性炭时,色素抑制率可达27.29%。以制得的螺旋藻肽为原料配制螺旋藻肽功能饮料。实验结果表明,在单因素实验的基础上,采用正交试验对饮料配制方法进行优化。对饮料品质影响的顺序依次为螺旋藻肽量、柠檬酸量、白砂糖量。获得的最佳配制方法为:每100mL螺旋藻肽饮料中添加螺旋藻肽0.5g,柠檬酸0.05g,白砂糖3g。
[Abstract]:As a food raw material, spirulina is widely used as a food raw material, and it is named as "human 21-century optimal health-care product" and "future super-nutritional food" (Zhang Wen et al.,2013). The content of the phycobiliprotein in the spirulina is as high as 50-70%. In addition, there is also a very desirable amino acid composition ratio, and the content of the eight amino acids necessary for the human body is close to or exceeds the standard specified by FAO. The spirulina has the effects of treating a series of diseases such as hypercholesterolemia, hyperglyceridemia, cardiovascular disease, inflammatory disease, cancer and viral infection. Spirulina can also be present as a functional ingredient in the food manufacturing industry (Hemamalini,2013). However, the application of the spirulina in the food manufacturing industry is limited by the disadvantages of poor dry powder solubility and heavy fishy smell. After the phycobiliprotein is treated by the protease, the physicochemical property is improved and the application range is expanded. Compared with the phycobiliprotein, the hydrolysate spirulina peptide has lower molecular weight and good physicochemical property, and is more easily digested and absorbed by the human body. But also has the physiological functions of improving the immune function of the human body, eliminating and preventing the body from being damaged by free radicals, comprehensively regulating the metabolism function of the human body, and the like, and the anti-oxidation capability of the human body is the most important. In this paper, the method of wall-breaking of spirulina is mainly studied, the process of preparing antioxidant peptide by single-enzyme method and double-enzymatic hydrolysis of spirulina is optimized, and the effect of the degree of hydrolysis on the foaming, emulsifying property and anti-oxidation ability of the spirulina peptide is studied. The production and control measures of the pigment in the enzymatic hydrolysis and the development of the functional beverage of the spirulina peptide were also studied. The mixing method, the ultrasonic crushing method, the repeated freeze-thaw method and the combination of the two can destroy the spirulina algae cells and lead the content to flow out. The results showed that the content of soluble protein in the cells was the most when the ultrasonic mode was the same, the ultrasonic power was 5%, the ultrasonic power was 7 s, and the interval was 3 s. The content of soluble protein in the thawed sample at 4 鈩，

本文编号：2497020