酶法改善大豆分离蛋白起泡性和乳化性的研究
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摘要
大豆蛋白是人类重要的营养来源和工业原料,价格低廉,不含胆固醇,但由于其功能性质例如起泡性与蛋清蛋白、乳化性与酪蛋白酸钠等天然优良性能的蛋白质相比还有很大的差距,导致其在现代食品加工中的应用受到一定限制。本文以大豆分离蛋白为原料,主要研究酶改性对其起泡性、乳化性以及结构性质的影响,研究对大豆分离蛋白的结构功能改造具有一定的学术意义,对于提高大豆蛋白的使用价值具有重要的现实意义。
论文首先研究了酶水解和酶交联并用的方法对于大豆分离蛋白起泡性以及乳化性的影响。论文创新性地设计通过控制底物的分子量和疏水性以提高大豆分离蛋白水解物交联产物的乳化性和起泡性的蛋白质改造方法。论文首先将大豆分离蛋白通过碱性蛋白酶alcalase水解,利用超滤进行分级获得不同分子量肽段(5kDa以下以及10kDa以上),利用谷氨酰胺转移酶(TGase)进行重组,测量其处理前后起泡性和乳化性的变化。实验结果显示大豆分离蛋白经alcalase水解并使用TGase交联后,产物起泡能力和泡沫稳定性有了显著的提高;但是乳化性略有不同,随着交联的不断进行,蛋白的乳化性逐渐增加,乳化稳定性略有下降。论文接着从疏水性角度研究了酶改性对大豆分离蛋白起泡性和乳化性的影响。同样将大豆分离蛋白进行水解,通过柱色谱将亲水性肽和疏水性肽分离,然后使用TGase进行交联,结果显示,将分子量1万以上的亲水性肽段和疏水性肽段以1:1混合后进行交联,产物的泡沫稳定性有显著提高。通过水解、按照分子量分级以及控制底物亲疏水性比例,酶交联后产物的起泡性和乳化性比原始大豆分离蛋白都有显著提高。
在此基础上,论文测定不同改造程度的大豆分离蛋白水解的分子量、分子量分布、表面疏水性、表面张力以及自由氨基含量等性质,初步探讨了酶改造对于大豆蛋白结构性质的影响,以及这种结构改变对于功能性质的影响。结果显示:酶处理显著提高了大豆分离蛋白的溶解性,改善了其表面疏水性和表面张力使得起泡和乳化过程更容易发生,自由氨基含量随着交联程度的提高而下降,分子量分布显示出了酶处理对大豆分离蛋白分子所做的改性。在结构性质与功能性质相关性方面,分子量分布对起泡能力的影响不显著,分子量在50kDa左右产生好的泡沫稳定性,且分布越集中其泡沫稳定性越好;乳化活性随着分子量的增加而变好,乳化稳定性与大豆分离蛋白相比变化不显著;高的氮溶指数是好的起泡性和乳化性的基础,在一定程度下,起泡能力随氮溶指数的升高而上升;起泡性和乳化性均随表面疏水性的增加而升高;起泡性与表面张力并未表现出显著的相关性,乳化性随着表面张力的升高而降低;泡沫稳定性和乳化稳定性表现出了随自由氨基升高而变好的趋势。
本论文的研究对大豆分离蛋白的结构改造以及应用的拓宽具有一定的指导意义和实用价值。
Soybean protein is not only an important protein source for human nutrition but also an industrial material with low price and low cholesterol level. However, its poor functionality limited its used in the food industry. For example, its foaming property is far worse than egg white protein. In this study, the influence of enzyme modification on the structure and functionality such as foaming property of soybean protein isolate was investigated。
SPI was hydrolyzed by alcalase, then the different molecular weight hydrolysates (MW<5kDa and MW>10kDa) were separated by ultrafiltration. Subsequently, the hydrolysates with proper molecular weights were cross-linked by TGase. In conclusion: the foaming capacity and foaming stability were improved significantly after the enzyme modification; the emulsifying activity was improved significantly but the emulsion stability was depressed.
Then the enzyme modification on the foam properties and the emulsifying properties of SPI was researched from the view of the hydrophilic-hydrophobic properties. The hydrophilic peptides and the hydrophobic were separated and cross-linked by TGase on different combination, then the foam properties and the emulsifying properties were measured. The results were the foam capacity and foam stability were improved but the emulsifying activity and the emulsion stability were depressed, this results were related to the change of the surface hydrophobicity and the content of amino group.
The structural properties (solubility, surface hydrophobicity, surface tension, the content of amino group,the distribution of molecular weight), the foam properties and the emulsifying properties were measured and be compared with the original SPI. the foam properties had obvious connection with solubility, surface hydrophobicity, surface tension and the distribution of molecular weight.
The relationship between the structural properties and foam properties and the emulsifying properties: the larger the molecular weight was, the better the foam stability and the emulsifying activity were; the higher the NSI was ,the better the foam capacity was; the lower the surface tension was, the better the emulsifying activity was; the more the free amino groups was, the foam stability and the emulsion stability were.
The research is meaningful to the structure modification and widely application of soybean protein isolate.
论文首先研究了酶水解和酶交联并用的方法对于大豆分离蛋白起泡性以及乳化性的影响。论文创新性地设计通过控制底物的分子量和疏水性以提高大豆分离蛋白水解物交联产物的乳化性和起泡性的蛋白质改造方法。论文首先将大豆分离蛋白通过碱性蛋白酶alcalase水解,利用超滤进行分级获得不同分子量肽段(5kDa以下以及10kDa以上),利用谷氨酰胺转移酶(TGase)进行重组,测量其处理前后起泡性和乳化性的变化。实验结果显示大豆分离蛋白经alcalase水解并使用TGase交联后,产物起泡能力和泡沫稳定性有了显著的提高;但是乳化性略有不同,随着交联的不断进行,蛋白的乳化性逐渐增加,乳化稳定性略有下降。论文接着从疏水性角度研究了酶改性对大豆分离蛋白起泡性和乳化性的影响。同样将大豆分离蛋白进行水解,通过柱色谱将亲水性肽和疏水性肽分离,然后使用TGase进行交联,结果显示,将分子量1万以上的亲水性肽段和疏水性肽段以1:1混合后进行交联,产物的泡沫稳定性有显著提高。通过水解、按照分子量分级以及控制底物亲疏水性比例,酶交联后产物的起泡性和乳化性比原始大豆分离蛋白都有显著提高。
在此基础上,论文测定不同改造程度的大豆分离蛋白水解的分子量、分子量分布、表面疏水性、表面张力以及自由氨基含量等性质,初步探讨了酶改造对于大豆蛋白结构性质的影响,以及这种结构改变对于功能性质的影响。结果显示:酶处理显著提高了大豆分离蛋白的溶解性,改善了其表面疏水性和表面张力使得起泡和乳化过程更容易发生,自由氨基含量随着交联程度的提高而下降,分子量分布显示出了酶处理对大豆分离蛋白分子所做的改性。在结构性质与功能性质相关性方面,分子量分布对起泡能力的影响不显著,分子量在50kDa左右产生好的泡沫稳定性,且分布越集中其泡沫稳定性越好;乳化活性随着分子量的增加而变好,乳化稳定性与大豆分离蛋白相比变化不显著;高的氮溶指数是好的起泡性和乳化性的基础,在一定程度下,起泡能力随氮溶指数的升高而上升;起泡性和乳化性均随表面疏水性的增加而升高;起泡性与表面张力并未表现出显著的相关性,乳化性随着表面张力的升高而降低;泡沫稳定性和乳化稳定性表现出了随自由氨基升高而变好的趋势。
本论文的研究对大豆分离蛋白的结构改造以及应用的拓宽具有一定的指导意义和实用价值。
Soybean protein is not only an important protein source for human nutrition but also an industrial material with low price and low cholesterol level. However, its poor functionality limited its used in the food industry. For example, its foaming property is far worse than egg white protein. In this study, the influence of enzyme modification on the structure and functionality such as foaming property of soybean protein isolate was investigated。
SPI was hydrolyzed by alcalase, then the different molecular weight hydrolysates (MW<5kDa and MW>10kDa) were separated by ultrafiltration. Subsequently, the hydrolysates with proper molecular weights were cross-linked by TGase. In conclusion: the foaming capacity and foaming stability were improved significantly after the enzyme modification; the emulsifying activity was improved significantly but the emulsion stability was depressed.
Then the enzyme modification on the foam properties and the emulsifying properties of SPI was researched from the view of the hydrophilic-hydrophobic properties. The hydrophilic peptides and the hydrophobic were separated and cross-linked by TGase on different combination, then the foam properties and the emulsifying properties were measured. The results were the foam capacity and foam stability were improved but the emulsifying activity and the emulsion stability were depressed, this results were related to the change of the surface hydrophobicity and the content of amino group.
The structural properties (solubility, surface hydrophobicity, surface tension, the content of amino group,the distribution of molecular weight), the foam properties and the emulsifying properties were measured and be compared with the original SPI. the foam properties had obvious connection with solubility, surface hydrophobicity, surface tension and the distribution of molecular weight.
The relationship between the structural properties and foam properties and the emulsifying properties: the larger the molecular weight was, the better the foam stability and the emulsifying activity were; the higher the NSI was ,the better the foam capacity was; the lower the surface tension was, the better the emulsifying activity was; the more the free amino groups was, the foam stability and the emulsion stability were.
The research is meaningful to the structure modification and widely application of soybean protein isolate.
引文
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2.冯昌友,陈建霞.大豆蛋白及其在食品工业中的应用[J].食品与机械,2000,2(76):21-22
3.杨凯,童正明,戴杏云.大豆分离蛋白的性质分析与改性研究[J].食品研究与开发,2006, 2(7):87-89
4. Naotoshi Matsudoml,T.S. Akio kato,Kunihiko Kobayashi. Conformation changes and functional properties of acid-modified soy protein[J].Journal of Agriculture and Food Chemistry,1985,49:1251-1256
5. K.L.Franzen.Functional properties of succinylated and acetylated soy protein[J]. Journal of Agriculture and Food Chemistry,1976,24 :788-795
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