【摘要】：小麥胚芽蛋白是以脫脂小麥胚芽為原料經(jīng)過堿溶酸沉等方式制備而成的蛋白質(zhì)產(chǎn)品,在食品、化工等領(lǐng)域具有重要的應(yīng)用前景。傳統(tǒng)的基于堿溶酸沉技術(shù)的提取工藝存在用水量高、提取率低和乳清廢水排放量大等一系列問題。此外,資源利用率和產(chǎn)品附加值較低。本文旨在以麥粕中蛋白質(zhì)為研究對(duì)象研究一種新型的堿溶酸沉提取小麥胚芽蛋白的方法,并在基于質(zhì)譜的蛋白質(zhì)種類識(shí)別的基礎(chǔ)上,研究了不同的提取條件對(duì)小麥胚芽中各種種類的蛋白質(zhì)的釋放行為造成的不同影響。通過小麥胚芽中試放大實(shí)驗(yàn)可將所得產(chǎn)品應(yīng)用到實(shí)際生產(chǎn)中,并通過改變提取溫度、溶劑的pH值及提取時(shí)間等因素可進(jìn)一步提高蛋白得率及蛋白含量,主要結(jié)論如下:1.基于HPLC-MS的蛋白質(zhì)識(shí)別方法研究,利用HPLC對(duì)小麥胚芽蛋白提取液進(jìn)行了分離,收集了分離出的主要蛋白質(zhì)組分,采用不同的蛋白酶進(jìn)行酶解的技術(shù)并結(jié)合了高效液相色譜串聯(lián)質(zhì)譜技術(shù)對(duì)高效液相分離出各種種類的蛋白質(zhì)進(jìn)行了一一識(shí)別。結(jié)果表明不同種類蛋白質(zhì)在色譜柱中保留時(shí)間的不同,總計(jì)識(shí)別出多種蛋白質(zhì),其中豐度較高的蛋白質(zhì)有17種。2.脫脂小麥胚芽蛋白質(zhì)釋放行為研究,在蛋白質(zhì)組分識(shí)別的基礎(chǔ)上,對(duì)麥粕中每一種蛋白質(zhì)的溶出過程進(jìn)行了檢測(cè),并對(duì)比了所有種類的蛋白質(zhì)在提取過程中的釋放速率以及釋放量的顯著變化。根據(jù)實(shí)驗(yàn)的結(jié)果可知,低分子量的蛋白釋放速率顯著快于高分子量蛋白質(zhì)的釋放速率,而且動(dòng)態(tài)提取的過程要比靜態(tài)提取的過程各組分蛋白的釋放速率要高很多,動(dòng)態(tài)提取過程中保留時(shí)間為18.78min的色譜峰中Chromosome3B蛋白釋放速率最快,提取2次的蛋白釋放率已達(dá)90%。3.基于小麥胚芽蛋白的酶解方法研究,考察了利用各種蛋白酶在不同的固液比、提取溫度、溶液pH值的條件下水解脫脂小麥胚芽蛋白制備得到小麥胚芽肽。比較木瓜蛋白酶、枯草桿菌蛋白酶,以及混合蛋白酶水解脫脂小麥胚芽蛋白的進(jìn)程曲線,結(jié)果顯示復(fù)合蛋白酶的酶解效果最好,其較佳作用條件為:底物濃度11.11%,pH 7.0,溫度45℃,酶與底物比1:200,時(shí)間為60 min。在此條件下,小麥胚芽蛋白的酶解效率最高為87%。此外,利用中試反應(yīng)釜,管式離心機(jī),噴霧干燥機(jī),冷凍干燥機(jī)等設(shè)備可將小麥胚芽蛋白的制備應(yīng)用到企業(yè)生產(chǎn),可制備出蛋白含量較高的小麥胚芽蛋白粉。本研究將利用堿溶酸沉技術(shù)應(yīng)用于小麥胚芽蛋白的提取,并考察了不同提取條件對(duì)蛋白收率及釋放速率的影響,同時(shí)對(duì)小麥胚芽蛋白的分離鑒定進(jìn)行優(yōu)化,對(duì)建立一種新型制備小麥胚芽蛋白的提取工藝具有重要的參考價(jià)值。
[Abstract]:Wheat germ protein is a protein product prepared from defatted wheat germ by alkaline acid precipitation. It has important application prospect in food, chemical industry and other fields. A series of problems such as high water consumption, low extraction rate and large discharge of whey wastewater exist in the traditional extraction process based on alkali soluble acid precipitation technology. In addition, resource utilization and product added value are low. The aim of this paper is to study a new method of extracting wheat germ protein from wheat meal by alkaline acid precipitation, and based on the recognition of protein species based on mass spectrometry. The effects of different extraction conditions on the release behavior of various kinds of proteins in wheat germ were studied. The obtained product can be applied to actual production through the pilot scale experiment of wheat germ, and the protein yield and protein content can be further improved by changing the extraction temperature, the pH value of solvent and the extraction time. The main conclusions are as follows: 1. Based on the study of protein recognition method based on HPLC-MS, wheat germ protein extract was separated by HPLC, and the main protein components were collected. Different protease hydrolysis techniques and high performance liquid chromatography-tandem mass spectrometry (HPLC / MS) were used to identify all kinds of proteins isolated by high performance liquid chromatography (HPLC). The results showed that the retention time of different kinds of proteins in the chromatographic column was different, and a total of many proteins were identified, of which 17 were found to have higher abundance. The protein release behavior of defatted wheat germ was studied. Based on the recognition of protein components, the dissolution process of each protein in wheat meal was determined. The release rate and release amount of all kinds of proteins were compared. According to the experimental results, the release rate of low molecular weight protein was significantly faster than that of high molecular weight protein, and the dynamic extraction process was much higher than that of static extraction process. The release rate of Chromosome3B protein was the fastest in the chromatographic peak with retention time of 18.78min during dynamic extraction, and the release rate of Chromosome3B protein reached 90.3in two times extraction. Based on the method of enzymatic hydrolysis of wheat germ protein, wheat germ peptide was prepared by hydrolysis of degreased wheat germ protein under different solid-liquid ratio, extraction temperature and pH value of solution. Compared with papain, Bacillus subtilis protease and mixed protease, the results showed that the enzymatic hydrolysis effect of complex protease was the best, and the optimum conditions were as follows: substrate concentration 11.11%. PH 7.0, temperature 45 鈩，

本文編號(hào)：2404098