芝麻渣中蛋白提取方法的比较研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-1130

摘要/Abstract

摘要：

芝麻渣是芝麻榨油后的主要副产物,含有丰富的蛋白质。但是目前除了少部分被当成饲料或肥料使用外,大部分芝麻渣被作为废弃物丢弃,造成环境污染和蛋白资源的严重浪费。将芝麻渣中的蛋白质提取进行高值化利用,具有广阔的应用前景和重要的现实意义。以芝麻渣为材料,分别利用低共熔溶剂（deep eutectic solvent,DES）法和碱溶酸沉法从芝麻渣中提取芝麻蛋白,建立了一种由醋酸钠（CH₃COONa·3H₂O,SAT）和尿素形成的DES提取芝麻蛋白的方法。考察了提取温度、时间、固液比和提取次数等条件对蛋白提取率的影响,通过单因素实验得到SAT-尿素体系提取芝麻蛋白的最佳工艺条件为：在温度为75℃,提取时间180 min、固液比为1∶50、利用乙醇沉淀蛋白的条件下,提取两次,总蛋白提取率可达71.41%,所制备的芝麻蛋白纯度为89.43%。相比于碱溶酸沉法,DES法不仅提取的芝麻蛋白纯度高,而且DES可回收使用、无废液污染。这些结果表明DES法是一种新型绿色的植物蛋白提取方法。

关键词: 低共熔溶剂, 芝麻蛋白, 绿色提取, 芝麻渣

Abstract:

Sesame residue is the main by-product of sesame oil processing,and contains rich proteins. Currently in addition to a few of them are used as feed or fertilizer,most of them is discarded as waste,which causes environmental pollution and serious waste of resources. Therefore,it has a broad application prospect and important practical significance to extract protein from sesame residue for high value utilization. In this study,sesame protein was extracted from sesame residues by the method of deep eutectic solvent（DES）and alkaline extraction followed by acid precipitation,respectively. A novel DES method of extracting sesame protein formed by sodium acetate（CH₃COONa·3H₂O,SAT）and urea was established. The effects of extraction temperature,time,solid-liquid ratio,and extraction times on the extraction rate of protein were investigated. The optimal extraction conditions of sesame protein by SAT-urea system were as follows：temperature 75℃,extraction time 180 min,solid-liquid ratio 1∶50,extracting twice under the conditions of using ethanol to precipitate proteins. Under the optimized conditions,the total protein extraction rate was 71.41%,and the purity of sesame protein was 89.43%. Compared with alkaline extraction followed by acid precipitation,the sesame protein extracted by DES presented high purity;moreover,DES can be recycled and used without waste pollution. These results show that DES extraction method is a new green method of extracting plant protein. This study provides a new insight for the green extraction of plant protein.

Key words: deep eutectic solvent, sesame protein, green extraction, sesame residues

焦晓波, 王高杨, 李锦红, 王晓艺, 孟国庆, 崔建东. 芝麻渣中蛋白提取方法的比较研究[J]. 生物技术通报, 2021, 37(4): 273-281.

JIAO Xiao-bo, WANG Gao-yang, LI Jin-hong, WANG Xiao-yi, MENG Guo-qing, CUI Jian-dong. A Comparative Study of Methods for Extracting Protein from Sesame Residue[J]. Biotechnology Bulletin, 2021, 37(4): 273-281.

图/表 13

表1 不同低共熔溶剂的制备条件

Table 1 Preparation conditions of different deep eutectic solvents

DES	氢键受体 Hydrogen bond receptor	氢键供体 Hydrogen bond donor	摩尔比 Molar ratio	制备温度 Temperature /℃
ChCl-乙酸 ChCl-acetic acid	氯化胆碱 ChCl	乙酸 Acetic acid	1﹕2	90
ChCl-尿素 ChCl- urea	氯化胆碱 ChCl	尿素 Urea	1﹕2	90
ChCl-葡萄糖 ChCl- glucose	氯化胆碱 ChCl	葡萄糖 Glucose	2﹕1	90
SAT-尿素 SAT-urea	醋酸钠 ChCl	尿素 Urea	1﹕2	90

图1 四种低共熔溶剂提取芝麻渣中的芝麻蛋白

Fig. 1 Extraction of sesame protein from sesame residue with 4 deep eutectic solvents

图2 不同固液比对SAT-尿素体系提取芝麻渣中蛋白的影响

Fig. 2 Effect of different solid-liquid ratios on the extraction of protein from sesame residue by SAT-urea system

图3 不同提取时间对SAT-尿素体系提取芝麻渣中蛋白的影响

Fig. 3 Effect of different extraction time on the extraction of protein from sesame residue by SAT-urea system

图4 不同温度对SAT-尿素体系提取芝麻渣中蛋白的影响

Fig. 4 Effect of different temperatures on the extraction of protein from sesame residue by SAT-urea system

表2 提取次数对提取率的影响

Table 2 Effect of extraction times on extraction rate

	第一次提取 First extraction	第二次提取 Second extraction	两次提取 Extraction twice
芝麻渣质量Quality of defatted sesame dreg/g	0.100	0.079	0.100
蛋白提取率Protein extraction rate/%	51.19	20.22	71.41

图5 SAT-尿素体系重复使用对提取芝麻渣中蛋白的影响

Fig. 5 Effect of the reusability of SAT-urea system on the extraction of protein from sesame residue

图6 氢氧化钠添加量对蛋白提取率的影响

Fig. 6 Effect of sodium hydroxide addition amount on protein extraction rate

图7 温度对蛋白提取率的影响

Fig. 7 Effect of temperature on protein extraction rate

图8 不同提取时间对蛋白提取率的影响

Fig. 8 Effect of different extraction time on protein extraction rate

图9 不同固液比对蛋白提取率的影响

Fig. 9 Effect of different solid-to-liquid ratios on protein extraction rate

图10 芝麻蛋白等电点测定

Fig. 10 Isoelectric point determination of sesame protein

表3 两种芝麻蛋白提取方法的比较

Table 3 Comparison of two extraction methods of sesame protein

提取方法 Extraction method	脱脂芝麻渣质量 Quality of defatted sesame dreg /g	蛋白提取率 Protein extraction rate /%	蛋白纯度 Protein purity /%	蛋白颜色 Color of protein
碱溶酸沉法 Alkali solution and acid precipitation method	0.100	73.86	83.375	深褐色 Dark brown
DES法 The DES method	0.100	71.41	89.433	浅黄色 Light yellow

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