麻疯籽制备生物柴油及提取蛋白质的技术研究
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摘要
麻疯树是一种具有很高的经济价值的树种,其根、茎、叶、果实均可被利用。其种子含油量能高达60%,油酸和亚油酸含量在70%以上,不饱和脂肪酸含量高,碘值较低,属半干性油,流动性好,是制备生物柴油的理想原料,利用麻疯籽油制备的生物柴油能达到欧II排放标准。麻疯籽中蛋白质含量丰富,占种仁重量的27%~32%,氨基酸组成也比较均衡,但由于麻疯籽毒蛋白(Curcin)和佛波醇酯的存在,榨油后的麻疯籽饼不能直接饲用,主要用作肥料。采用适当的蛋白质提取工艺,探索去除麻疯籽蛋白中毒素的方法,将有利于麻疯籽饼粕的开发利用。
本文就以下几个方面对麻疯籽油和麻疯籽蛋白进行了研究:
1.本课题首先进行了冷榨—浸出法制取麻疯籽油的研究,通过试验分析,认为在压榨压力40MPa,压榨时间120min,温度65℃,原料含水量10%条件下进行冷榨试验,出油效果较好,出油率约为41%,冷榨饼残油量为28%。利用正己烷为溶剂在溶剂比1.2:1,温度55℃,时间30min,连续浸出冷榨饼5次的条件下,最终麻疯籽粕残油量低至0.8%。
2.经过水化脱胶、真空脱水后的麻疯籽油酸值为15.49mgKOH/g油,而均相碱催化制备生物柴油,要求原料油酸值≤1.0mgKOH/g油,因此需要进行降酸处理。本文分别就溶剂法萃取脱酸和浓硫酸催化预酯化降酸两种方法进行了研究。通过单因素试验和响应面分析,得出麻疯籽油溶剂法脱酸的最优工艺:醇油比(V/V油)2.0:1,麻疯籽油含水量0.08%,萃取温度45℃,连续萃取5次,每次萃取10min。而浓硫酸催化预酯化降酸通过添加助溶剂四氢呋喃,经过单因素试验和正交试验分析,得出最优工艺为:甲醇添加量(V/V油)0.85:1,反应时间45min,反应温度65℃,催化剂添加量(V/V油)0.012:1,助溶剂添加量(V/V油)0.5:1。
3.为选择合适的助溶剂来有效促进甲醇和麻疯籽油形成均相体系。本文通过对8种溶剂的性质对比,以及在不同的温度条件下,乙醚或四氢呋喃溶剂与麻疯籽油、甲醇形成的三元相图的比较,最终选择四氢呋喃作为本次研究的助溶剂。
4.在浓硫酸催化预酯化有效降酸的条件下,本文采用以四氢呋喃为助溶剂,NaOH为催化剂,进行了麻疯籽油制备生物柴油的甲酯化研究,通过单因素分析和正交试验设计,认为在甲醇添加量(V/V油)0.56:1,反应时间18min,反应温度65℃,催化剂添加量(w/w油)1%,助溶剂添加量(V/V油)0.5:1的条件下,甲酯转化率较高。
5.本文采用碱提酸沉法对麻疯籽粕中的麻疯籽蛋白进行了提取研究。通过单因素试验研究各因素对提取率的影响,应用正交试验优化工艺条件。得到的最优工艺条件为:碱提料液比1:11,pH 8.5,温度55℃,时间100min;酸沉pH 4.4,温度55℃,时间40min。在该工艺条件下,麻疯籽蛋白提取率达到76.63%,分离物蛋白纯度达到81.18%。
Jatropha curcas is a kind of tree that has very high economic value, its roots, stems, leaves and fruits all can be used.The oil content of its seed is as high as 60%, in which the ratio of oleic acid and linoleic acid is more than 70%,the content of unsaturated fatty acids are high whereas the iodine value is low. It is a semi-dry oil with good fluidity, which is the ideal raw material for biodiesel. The biodiesel prepared by Jatropha curcas oil can meet the II emission standards of European.The content of protein in Jatropha curcas seed is very rich, which accounts for 27% ~ 32% of seed weight, and the amino acid composition is balanced. However, the presence of Curcin and phorbol ester, the seed cake after oil extraction can not directly feed, it’s mainly used as fertilizer.Therefore, using appropriate protein extraction technology to explore the way of removing toxin in Jatropha curcas seeds’protein will help the development and utilization of Jatropha curcas seed cake.
In this paper, the following aspects of Jatropha oil and its seed protein were studied:
(1) This study began with research on Jatropha curcas seed oil extraction by cold-pressed–solvent extraction, after experiment and analysis, the better conditions were as follows: the pressure 40MPa, time 120min, temperature 65℃, 10% water content. Under the Conditions, the oil extraction was around 41%, while the residual oil in cold pressed cake was about 28%.Using hexane as solvent, under the conditions: the solvent ratio 1.2, temperature 55℃, time 30min, continuous extracting the cold pressed cake 5 times, the final residual oil in Jatropha curcas seeds was as low as 0.8%.
(2) After hydration degumming and vacuum dehydration, the acid value of Jatropha curcas seed oil was 15.49 mg KOH/g oil, while the acid value of raw materials for biodiesel preparation using homogeneous base catalyst is required lower than 1.0 mg KOH/g oil, thus there is a need to reduce acid value. This paper studied on the solvent extraction deacidifieing as well as acid-dropping by pre-esterification catalyzed by sulfuric acid. Through single factor experiments and response surface analysis, the optimal solvent deacidifieing process was obtained: alcohol to oil ratio (V/V oil) 2.0:1, water content in Jatropha curcas seed oil 0.08%, extraction temperature 45℃, extracted 5 times for each of 10min.While, as for the method of sulfuric acid pre-esterification deacidifieing, by adding THF as a cosolvent, through single factor experiments and orthogonal analysis, the optimum conditions were obtained as follows: amount of methanol added (V/V oil) 0.85:1, reaction time 45min, reaction temperature 65℃, catalyst loadings (V/V oil) 0.012:1, addition of cosolvent (V/V oil) 0.5:1.
(3) In order to select the appropriate solvent that can effectively promote the formation of methanol and Jatropha curcas oil homogeneous, the comparisons of the nature contrast of 8 solvent and the ternary phase diagram that formed by the ether or tetrahydrofuran solvent and Jatropha curcas seed oil, methanol in different temperatures were studied, we finally chosed THF as a cosolvent.
(4) In the conditions when the acid value was effectively reduced by sulfuric acid pre- esterification, we used THF as a cosolvent and NaOH as catalyst, studied the biodiesel process of Jatropha curcas seed oil through methyl esterification. By single factor analysis and orthogonal experimental design, when the amount of methanol added (V/V oil)was 0.56:1, reaction time 18min, reaction temperature 65℃, catalyst loadings (w/w oil) 1% addition of cosolvent (V/V oil) 0.5:1 conditions, a higher conversion rate of methyl esterification was obtained.
(5) In this paper, alkaline extraction and acid precipitation were employed to extract protein from Jatropha curcas seed meal.After respectively studied the influence of different factors on extraction yield by single factor experiments, and then optimized conditions by orthogonal test, the optimized conditions were obtained as follows: alkaline extraction solid-liquid ratio 1:11, pH 8.5, temperature 55℃, time 100min;acid pH 4.4, temperature 55℃, time 40min.Under the process conditions, the separation yield of Jatropha curcas seed protein was 76.63%, purity of protein isolated reached 81.18%.
本文就以下几个方面对麻疯籽油和麻疯籽蛋白进行了研究:
1.本课题首先进行了冷榨—浸出法制取麻疯籽油的研究,通过试验分析,认为在压榨压力40MPa,压榨时间120min,温度65℃,原料含水量10%条件下进行冷榨试验,出油效果较好,出油率约为41%,冷榨饼残油量为28%。利用正己烷为溶剂在溶剂比1.2:1,温度55℃,时间30min,连续浸出冷榨饼5次的条件下,最终麻疯籽粕残油量低至0.8%。
2.经过水化脱胶、真空脱水后的麻疯籽油酸值为15.49mgKOH/g油,而均相碱催化制备生物柴油,要求原料油酸值≤1.0mgKOH/g油,因此需要进行降酸处理。本文分别就溶剂法萃取脱酸和浓硫酸催化预酯化降酸两种方法进行了研究。通过单因素试验和响应面分析,得出麻疯籽油溶剂法脱酸的最优工艺:醇油比(V/V油)2.0:1,麻疯籽油含水量0.08%,萃取温度45℃,连续萃取5次,每次萃取10min。而浓硫酸催化预酯化降酸通过添加助溶剂四氢呋喃,经过单因素试验和正交试验分析,得出最优工艺为:甲醇添加量(V/V油)0.85:1,反应时间45min,反应温度65℃,催化剂添加量(V/V油)0.012:1,助溶剂添加量(V/V油)0.5:1。
3.为选择合适的助溶剂来有效促进甲醇和麻疯籽油形成均相体系。本文通过对8种溶剂的性质对比,以及在不同的温度条件下,乙醚或四氢呋喃溶剂与麻疯籽油、甲醇形成的三元相图的比较,最终选择四氢呋喃作为本次研究的助溶剂。
4.在浓硫酸催化预酯化有效降酸的条件下,本文采用以四氢呋喃为助溶剂,NaOH为催化剂,进行了麻疯籽油制备生物柴油的甲酯化研究,通过单因素分析和正交试验设计,认为在甲醇添加量(V/V油)0.56:1,反应时间18min,反应温度65℃,催化剂添加量(w/w油)1%,助溶剂添加量(V/V油)0.5:1的条件下,甲酯转化率较高。
5.本文采用碱提酸沉法对麻疯籽粕中的麻疯籽蛋白进行了提取研究。通过单因素试验研究各因素对提取率的影响,应用正交试验优化工艺条件。得到的最优工艺条件为:碱提料液比1:11,pH 8.5,温度55℃,时间100min;酸沉pH 4.4,温度55℃,时间40min。在该工艺条件下,麻疯籽蛋白提取率达到76.63%,分离物蛋白纯度达到81.18%。
Jatropha curcas is a kind of tree that has very high economic value, its roots, stems, leaves and fruits all can be used.The oil content of its seed is as high as 60%, in which the ratio of oleic acid and linoleic acid is more than 70%,the content of unsaturated fatty acids are high whereas the iodine value is low. It is a semi-dry oil with good fluidity, which is the ideal raw material for biodiesel. The biodiesel prepared by Jatropha curcas oil can meet the II emission standards of European.The content of protein in Jatropha curcas seed is very rich, which accounts for 27% ~ 32% of seed weight, and the amino acid composition is balanced. However, the presence of Curcin and phorbol ester, the seed cake after oil extraction can not directly feed, it’s mainly used as fertilizer.Therefore, using appropriate protein extraction technology to explore the way of removing toxin in Jatropha curcas seeds’protein will help the development and utilization of Jatropha curcas seed cake.
In this paper, the following aspects of Jatropha oil and its seed protein were studied:
(1) This study began with research on Jatropha curcas seed oil extraction by cold-pressed–solvent extraction, after experiment and analysis, the better conditions were as follows: the pressure 40MPa, time 120min, temperature 65℃, 10% water content. Under the Conditions, the oil extraction was around 41%, while the residual oil in cold pressed cake was about 28%.Using hexane as solvent, under the conditions: the solvent ratio 1.2, temperature 55℃, time 30min, continuous extracting the cold pressed cake 5 times, the final residual oil in Jatropha curcas seeds was as low as 0.8%.
(2) After hydration degumming and vacuum dehydration, the acid value of Jatropha curcas seed oil was 15.49 mg KOH/g oil, while the acid value of raw materials for biodiesel preparation using homogeneous base catalyst is required lower than 1.0 mg KOH/g oil, thus there is a need to reduce acid value. This paper studied on the solvent extraction deacidifieing as well as acid-dropping by pre-esterification catalyzed by sulfuric acid. Through single factor experiments and response surface analysis, the optimal solvent deacidifieing process was obtained: alcohol to oil ratio (V/V oil) 2.0:1, water content in Jatropha curcas seed oil 0.08%, extraction temperature 45℃, extracted 5 times for each of 10min.While, as for the method of sulfuric acid pre-esterification deacidifieing, by adding THF as a cosolvent, through single factor experiments and orthogonal analysis, the optimum conditions were obtained as follows: amount of methanol added (V/V oil) 0.85:1, reaction time 45min, reaction temperature 65℃, catalyst loadings (V/V oil) 0.012:1, addition of cosolvent (V/V oil) 0.5:1.
(3) In order to select the appropriate solvent that can effectively promote the formation of methanol and Jatropha curcas oil homogeneous, the comparisons of the nature contrast of 8 solvent and the ternary phase diagram that formed by the ether or tetrahydrofuran solvent and Jatropha curcas seed oil, methanol in different temperatures were studied, we finally chosed THF as a cosolvent.
(4) In the conditions when the acid value was effectively reduced by sulfuric acid pre- esterification, we used THF as a cosolvent and NaOH as catalyst, studied the biodiesel process of Jatropha curcas seed oil through methyl esterification. By single factor analysis and orthogonal experimental design, when the amount of methanol added (V/V oil)was 0.56:1, reaction time 18min, reaction temperature 65℃, catalyst loadings (w/w oil) 1% addition of cosolvent (V/V oil) 0.5:1 conditions, a higher conversion rate of methyl esterification was obtained.
(5) In this paper, alkaline extraction and acid precipitation were employed to extract protein from Jatropha curcas seed meal.After respectively studied the influence of different factors on extraction yield by single factor experiments, and then optimized conditions by orthogonal test, the optimized conditions were obtained as follows: alkaline extraction solid-liquid ratio 1:11, pH 8.5, temperature 55℃, time 100min;acid pH 4.4, temperature 55℃, time 40min.Under the process conditions, the separation yield of Jatropha curcas seed protein was 76.63%, purity of protein isolated reached 81.18%.
引文
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