大豆糖蜜中低聚糖的分离纯化
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摘要
本论文研究了以大豆糖蜜为原料,采用超滤技术、大孔吸附树脂法、离子交换树脂法提取大豆低聚糖的工艺。
首先对预处理前后大豆糖蜜的主要成分和性质进行了测定。经预处理后得到了透光率为88.1%,基本澄清的糖蜜上清液,其中总糖占了60.48%。通过单因素实验研究了超滤实验中pH值、温度对膜通量、总糖透过率和大分子类蛋白截留率的影响。所用的超滤膜截留相对分子质量(MWCO)为10000,采用间断全过滤的操作方式,确定的最佳超滤工艺为:pH7.0、T:20℃、压力0.6MPa(膜允许的标准操作压力范围)。在此条件下,总糖透过率为75.65%,大分子类蛋白完全截留,超滤液的透光率为93.5%。接着研究了3种不同的大孔树脂对超滤液的脱色效果,结果表明AB-8树脂对超滤液的脱色率最高,因此选择了AB-8大孔吸附树脂作为脱色用树脂。通过单因素实验进一步研究了pH值、流速、上样量对超滤液的脱色效果的影响,确定了AB-8树脂的最佳脱色条件为:室温、pH4.0,流速1.5BV/h,上样量4BV。此条件下脱色率可达90%以上,总糖损失率为10%左右。以70%的乙醇水溶液作为洗脱剂,解吸率可达89.69%。
最后,对大豆低聚糖粗糖液中存在的主要杂质进行了分析,并比较了纳滤和离子交换树脂法对粗糖液纯化的效果,最终选择了离子交换技术,选用了001×7型强酸性阳离子交换树脂与D301型弱碱性阴离子交换树脂,按先阳后阴的方式连接。具体的操作条件为:室温、流速1.2BV/h。此条件下001×7型树脂的交换能力约为11BV,D301型树脂的交换能力为7BV左右。最终得到的糖液的电导率与生活用水十分接近,几乎所有的含氮物质及绝大部分的酚类物质得到了去除。
经过以上的分离纯化,得到了浅黄色、澄清透亮的大豆低聚糖糖浆。该产品的各项指标为:固形物66.73%,蔗糖53.43%,棉子糖6.90%,水苏糖14.92%,灰分1.02%,无含氮物质检出。
This paper was studied on using ultrafiltration technology, macroporous resins and ion exchange resins to extract soybean oligosaccharides from soy molasses.
Firstly, the components and characteristics of soy molasses both before and after pretreating were determined. The transmittance of the supernatant was 88.1%, this supernatant was exactly clean and clear, the total sugar accounted for 60.48%. During the ultrafiltration process, the effect of pH value, temperature on membrane flux, the total sugar permeation rate and the cut-off rate of macromolecule proteins were studied through the single factor experiments. The optimum conditions were: 10000 molecular weight cut-off membrane, using diafiltration operating mode, pH7.0, 20℃. Under these conditions, almost all of the macromolecule proteins were removed, and about 75.65% of the oligosaccharide was retained, the transmittance of the UF permeate was 93.5%.
Secondly, the decoloration effect of three types of macroporous resins on UF permeate was studied. AB-8 resin was chosen as decolorizing resin by comparing the decoloration. The suitable operation conditions were: room temperature, pH4.0, work solution speed of 1.5BV/h, the handling capacity was 4BV. Under this condition, the decoloration rate was over 90%, the total sugar loss was about 10%. The 70% aqueous alcohol was used as desorption reagent, the desorption rate reached 89.69%.
At last, the main impurities existed in the crude soybean oligosaccharides were analysised and the purification effect both by nanofiltration and ion exchange resins on it were compared. the ion exchange technology was chosen ultimately. 001×7 was chosen as the cation resin, and D301 was chosen as anion resin. The operation condition was: room temperature, work solution speed of 1.2BV/h. The conductivity of the purified soybean oligosaccharides was very close to the tap water, nearly all of the nitrogenous material and most of the phenolic material was removed.
After separation and purification, a pale yellow, translucent and clear soybean oligosaccharides syrup was prepared. The basic components of the end-product were: 66.73% solid, among this sucrose, raffinose, stachyose accounted for 53.43%, 6.90%, 14.92% respectively, ash 1.02%, no nitrogenous material was detected.
首先对预处理前后大豆糖蜜的主要成分和性质进行了测定。经预处理后得到了透光率为88.1%,基本澄清的糖蜜上清液,其中总糖占了60.48%。通过单因素实验研究了超滤实验中pH值、温度对膜通量、总糖透过率和大分子类蛋白截留率的影响。所用的超滤膜截留相对分子质量(MWCO)为10000,采用间断全过滤的操作方式,确定的最佳超滤工艺为:pH7.0、T:20℃、压力0.6MPa(膜允许的标准操作压力范围)。在此条件下,总糖透过率为75.65%,大分子类蛋白完全截留,超滤液的透光率为93.5%。接着研究了3种不同的大孔树脂对超滤液的脱色效果,结果表明AB-8树脂对超滤液的脱色率最高,因此选择了AB-8大孔吸附树脂作为脱色用树脂。通过单因素实验进一步研究了pH值、流速、上样量对超滤液的脱色效果的影响,确定了AB-8树脂的最佳脱色条件为:室温、pH4.0,流速1.5BV/h,上样量4BV。此条件下脱色率可达90%以上,总糖损失率为10%左右。以70%的乙醇水溶液作为洗脱剂,解吸率可达89.69%。
最后,对大豆低聚糖粗糖液中存在的主要杂质进行了分析,并比较了纳滤和离子交换树脂法对粗糖液纯化的效果,最终选择了离子交换技术,选用了001×7型强酸性阳离子交换树脂与D301型弱碱性阴离子交换树脂,按先阳后阴的方式连接。具体的操作条件为:室温、流速1.2BV/h。此条件下001×7型树脂的交换能力约为11BV,D301型树脂的交换能力为7BV左右。最终得到的糖液的电导率与生活用水十分接近,几乎所有的含氮物质及绝大部分的酚类物质得到了去除。
经过以上的分离纯化,得到了浅黄色、澄清透亮的大豆低聚糖糖浆。该产品的各项指标为:固形物66.73%,蔗糖53.43%,棉子糖6.90%,水苏糖14.92%,灰分1.02%,无含氮物质检出。
This paper was studied on using ultrafiltration technology, macroporous resins and ion exchange resins to extract soybean oligosaccharides from soy molasses.
Firstly, the components and characteristics of soy molasses both before and after pretreating were determined. The transmittance of the supernatant was 88.1%, this supernatant was exactly clean and clear, the total sugar accounted for 60.48%. During the ultrafiltration process, the effect of pH value, temperature on membrane flux, the total sugar permeation rate and the cut-off rate of macromolecule proteins were studied through the single factor experiments. The optimum conditions were: 10000 molecular weight cut-off membrane, using diafiltration operating mode, pH7.0, 20℃. Under these conditions, almost all of the macromolecule proteins were removed, and about 75.65% of the oligosaccharide was retained, the transmittance of the UF permeate was 93.5%.
Secondly, the decoloration effect of three types of macroporous resins on UF permeate was studied. AB-8 resin was chosen as decolorizing resin by comparing the decoloration. The suitable operation conditions were: room temperature, pH4.0, work solution speed of 1.5BV/h, the handling capacity was 4BV. Under this condition, the decoloration rate was over 90%, the total sugar loss was about 10%. The 70% aqueous alcohol was used as desorption reagent, the desorption rate reached 89.69%.
At last, the main impurities existed in the crude soybean oligosaccharides were analysised and the purification effect both by nanofiltration and ion exchange resins on it were compared. the ion exchange technology was chosen ultimately. 001×7 was chosen as the cation resin, and D301 was chosen as anion resin. The operation condition was: room temperature, work solution speed of 1.2BV/h. The conductivity of the purified soybean oligosaccharides was very close to the tap water, nearly all of the nitrogenous material and most of the phenolic material was removed.
After separation and purification, a pale yellow, translucent and clear soybean oligosaccharides syrup was prepared. The basic components of the end-product were: 66.73% solid, among this sucrose, raffinose, stachyose accounted for 53.43%, 6.90%, 14.92% respectively, ash 1.02%, no nitrogenous material was detected.
引文
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[3] N Qureshi, A Lolas, HP Blaschek. Soy molasses as fermentation substrate for production of butanol using Clostridium beijerinckii BA101[J]. Journal of Industrial Microbiology & Biotechnology, 2001, 26: 290-295.
[4] Daniel K. Y. Solaiman, Richard D. Ashby, Alberto Nunez, Thomas A. Foglia. Production of sophorolipids by candida bombicola grown on soy molasses as substrate[J]. Biotechnology Letters, 2004, 26: 1241-1245.
[5] Montelongo J. L., Chassy B. M., McCord J. D. Lactobacillus salivarius for conversion of soy molasses into Lactic Acid[J]. Journal of Food Science. 1993, 58 (4): 863-866.
[6] Gugger, Eric T., Dueppen, etal. Production of isoflavone enriched fractions from soy protein extracts[P]. US Patent: 5792503, 1998.
[7]方伟辉,华欲飞,张喆等.大豆粕酒精可溶物的成分分离与鉴定[J].中国油脂, 2004, 29(1): 57-61.
[8]金其荣,徐勤.大豆低聚糖生产、生理功能及其应用[J].食品科学, 1994, 11: 7– 12.
[9] You-Jin J, Se-Kwon K. Production of chitooligosaccharides using an ultrafiltration membrance reactor and their antibacterial activity[J]. Carbohydrate polymers, 2000, 41: 133-141.
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