大蒜中三种生物活性物质提取与应用研究
摘要
大蒜是重要的药食兼用的植物资源之一,含有丰富的大蒜多糖、蒜氨酸酶、SOD等活性成分,已有的研究证实其具有增强免疫力、防病治病、延缓衰老等重要生理功能,且在食品、药品和化妆品等领域中有广阔的应用潜力,但目前未见以大蒜为原料工业化生产这三种活性成分及有关产品的报道。为此,本论文研究这三种活性成分的有关性质、提取和应用等问题,以为工业化生产提供依据。获得了如下主要研究结果:
①采用硫酸铵盐析法分步分离大蒜中蒜氨酸酶和SOD。蒜氨酸酶在40%,SOD在90%硫酸铵饱和度时最多。经过盐析、透析步骤后,获得的蒜氨酸酶和SOD比活力分别为:20.4U/mg、127.98 U/mg,活力回收分别为:73%、41%。
②确定了综合提取蒜氨酸酶和大蒜多糖的工业化生产技术。先将大蒜打浆,过滤除渣,然后调节澄清大蒜汁pH值至4,离心、沉淀用磷酸盐缓冲液溶解后得蒜氨酸酶,离心后的上清液过大孔树脂柱,得到大蒜多糖精制液,干燥得到大蒜多糖。该工艺大蒜多糖提取率为85%,蒜氨酸酶的比活为:17U/mg,活力回收62%。该技术适合蒜氨酸酶和大蒜多糖的工业化生产。
③利用MALDI-TOF MS技术测定酒精分级沉淀大蒜多糖的分子量分布特征。不同浓度酒精分级沉淀大蒜多糖时,多糖或寡聚糖的聚合度随着酒精浓度提高而下降。当酒精浓度为70%、80%、90%、95%时,沉淀的大蒜多糖以寡聚糖为主,基峰聚合度分别为7、7、4、6,最大聚合度分别为16、18、17、12。
④通过发酵实验证实了大蒜多糖可部分被乳酸菌发酵。大蒜多糖有增值益生菌的效果,益生菌可能主要发酵低聚合度的寡聚果糖。益尘菌发酵不同浓度酒精分级沉淀所获的大蒜多糖时,随酒精浓度越高,所获大蒜多糖的可发酵百分率越高。当酒精浓度分别为70%、80%、90%、95%时,分级沉淀大蒜多糖的可发酵率分别为:4.5%、9.1%、27.8%、55.6%。
⑤确定了以新鲜大蒜为原料生产大蒜多糖饮料的生产工艺和配方。以新鲜大蒜为原料,采用硅溶胶-膨润土法除蛋白后,大蒜多糖中的蛋白质含量为0.014 mg/ml;以饮用水加入2%的大蒜多糖、0.04%安赛蜜、0.004%糖精和0.12%乳酸时,可调配出一种天然、保健的多糖饮料,且大蒜多糖饮料的口感最佳。
Garlic is widely used both as food ingredient and herbal medicine in the world,which is rich in garlic polysaccharide,Alliinase and SOD.Physiological functions such as immunity regulatory function,delaying Senescence and preventing and curing diseases, have been proved by scientific research.Garlic polysaccharide,Alliinase and SOD have great potential applications in the field of food,medicine and cosmetic.However,there is no enterprise produces garlic functional products.We mainly made researches on the three active constituents' properties,extracting and intensive processing methods,which would provide a significant basis for mass industrial production.The research results include:
①While extracting and preliminarily purifying SOD and Alliinase from the garlic, 40%and 90%saturation was tested as the best point for two-step ammonium sulfate salting-out of the Alliinase and SOD.After the processing of salting-out and dialysis,the activity of Alliinase and SOD is 72U/mg,127.98 U/mg,and the recovery rate is 73%、41% respectively.
②The technology of extracting garlic polysaccharide and Alliinase was introduced in this paper:First,the garlic pulping,filtering,modifing pH value to 4,centrifugalizing;then the Alliinase was obtained after sediment was dissolve in phosphate buffer.The supernatant fluid after centrifugal procedure was purified by the Macroporous Adsorbed Resin,the fined garlic polysaccharide was gained and the extraction ratio is 85%.The Alliinase activity of is 72U/mg,and the recovery rate is 62%.This technology used a batch of garlic produce Alliinase and garlic polysaccharide two valuable active constituents,has raised the utilization rate of garlic resource greatly and reduced manufacturing cost.The results could be widely used in the commercial production of garlic polysaccharide and Alliinase.
③Distribution and molecular weight of garlic polysaccharide which was fractionated by the alcohol fractional precipitation technique have been measured by MALDI-TOF MS. With the increase of alcohol concentration,the degree of molecular weight of garlic polysaccharide revealed a trend of gradual decrease,when the alcohol concentration was 70%,80%,90%and 95%,the base peak degree of Polymerization were 7,7,4 and 6,the maximum degree of Polymerization were 16,18,17 and 12 respectively.
④Our research confirmed that the garlic polysaccharide can be partly fermented by the Lactobacillus.The garlic polysaccharide has the effect of promoting the growth of probiotics,which mainly ferment oligosaccharide.Experimental results indicated with the increase of alcohol concentration,the degree of the fermentation raised.When the alcohol concentration were 70%,80%,90%and 95%,the fermenting rate of garlic polysaccharide were 4.5%,9.1%,27.8%and 55.6%respectively.
⑤The research and development of garlic polysaccharide drinks mainly discussed the best processing technology and dispensing of the garlic polysaccharide functional drinks. The content of the remaining protein is 0.014 mg/ml in the garlic polysaccharide drink by the removing protein technology of the acid silica sol-Bentonite.And it proved that the optimum parameters were:2%garlic polysaccharide,0.04%Acesulfame-k,0.004%sodium saccharin,0.12%Lactic acid,which provides a experimental model for producing garlic polysaccharide functional drink.
①采用硫酸铵盐析法分步分离大蒜中蒜氨酸酶和SOD。蒜氨酸酶在40%,SOD在90%硫酸铵饱和度时最多。经过盐析、透析步骤后,获得的蒜氨酸酶和SOD比活力分别为:20.4U/mg、127.98 U/mg,活力回收分别为:73%、41%。
②确定了综合提取蒜氨酸酶和大蒜多糖的工业化生产技术。先将大蒜打浆,过滤除渣,然后调节澄清大蒜汁pH值至4,离心、沉淀用磷酸盐缓冲液溶解后得蒜氨酸酶,离心后的上清液过大孔树脂柱,得到大蒜多糖精制液,干燥得到大蒜多糖。该工艺大蒜多糖提取率为85%,蒜氨酸酶的比活为:17U/mg,活力回收62%。该技术适合蒜氨酸酶和大蒜多糖的工业化生产。
③利用MALDI-TOF MS技术测定酒精分级沉淀大蒜多糖的分子量分布特征。不同浓度酒精分级沉淀大蒜多糖时,多糖或寡聚糖的聚合度随着酒精浓度提高而下降。当酒精浓度为70%、80%、90%、95%时,沉淀的大蒜多糖以寡聚糖为主,基峰聚合度分别为7、7、4、6,最大聚合度分别为16、18、17、12。
④通过发酵实验证实了大蒜多糖可部分被乳酸菌发酵。大蒜多糖有增值益生菌的效果,益生菌可能主要发酵低聚合度的寡聚果糖。益尘菌发酵不同浓度酒精分级沉淀所获的大蒜多糖时,随酒精浓度越高,所获大蒜多糖的可发酵百分率越高。当酒精浓度分别为70%、80%、90%、95%时,分级沉淀大蒜多糖的可发酵率分别为:4.5%、9.1%、27.8%、55.6%。
⑤确定了以新鲜大蒜为原料生产大蒜多糖饮料的生产工艺和配方。以新鲜大蒜为原料,采用硅溶胶-膨润土法除蛋白后,大蒜多糖中的蛋白质含量为0.014 mg/ml;以饮用水加入2%的大蒜多糖、0.04%安赛蜜、0.004%糖精和0.12%乳酸时,可调配出一种天然、保健的多糖饮料,且大蒜多糖饮料的口感最佳。
Garlic is widely used both as food ingredient and herbal medicine in the world,which is rich in garlic polysaccharide,Alliinase and SOD.Physiological functions such as immunity regulatory function,delaying Senescence and preventing and curing diseases, have been proved by scientific research.Garlic polysaccharide,Alliinase and SOD have great potential applications in the field of food,medicine and cosmetic.However,there is no enterprise produces garlic functional products.We mainly made researches on the three active constituents' properties,extracting and intensive processing methods,which would provide a significant basis for mass industrial production.The research results include:
①While extracting and preliminarily purifying SOD and Alliinase from the garlic, 40%and 90%saturation was tested as the best point for two-step ammonium sulfate salting-out of the Alliinase and SOD.After the processing of salting-out and dialysis,the activity of Alliinase and SOD is 72U/mg,127.98 U/mg,and the recovery rate is 73%、41% respectively.
②The technology of extracting garlic polysaccharide and Alliinase was introduced in this paper:First,the garlic pulping,filtering,modifing pH value to 4,centrifugalizing;then the Alliinase was obtained after sediment was dissolve in phosphate buffer.The supernatant fluid after centrifugal procedure was purified by the Macroporous Adsorbed Resin,the fined garlic polysaccharide was gained and the extraction ratio is 85%.The Alliinase activity of is 72U/mg,and the recovery rate is 62%.This technology used a batch of garlic produce Alliinase and garlic polysaccharide two valuable active constituents,has raised the utilization rate of garlic resource greatly and reduced manufacturing cost.The results could be widely used in the commercial production of garlic polysaccharide and Alliinase.
③Distribution and molecular weight of garlic polysaccharide which was fractionated by the alcohol fractional precipitation technique have been measured by MALDI-TOF MS. With the increase of alcohol concentration,the degree of molecular weight of garlic polysaccharide revealed a trend of gradual decrease,when the alcohol concentration was 70%,80%,90%and 95%,the base peak degree of Polymerization were 7,7,4 and 6,the maximum degree of Polymerization were 16,18,17 and 12 respectively.
④Our research confirmed that the garlic polysaccharide can be partly fermented by the Lactobacillus.The garlic polysaccharide has the effect of promoting the growth of probiotics,which mainly ferment oligosaccharide.Experimental results indicated with the increase of alcohol concentration,the degree of the fermentation raised.When the alcohol concentration were 70%,80%,90%and 95%,the fermenting rate of garlic polysaccharide were 4.5%,9.1%,27.8%and 55.6%respectively.
⑤The research and development of garlic polysaccharide drinks mainly discussed the best processing technology and dispensing of the garlic polysaccharide functional drinks. The content of the remaining protein is 0.014 mg/ml in the garlic polysaccharide drink by the removing protein technology of the acid silica sol-Bentonite.And it proved that the optimum parameters were:2%garlic polysaccharide,0.04%Acesulfame-k,0.004%sodium saccharin,0.12%Lactic acid,which provides a experimental model for producing garlic polysaccharide functional drink.
引文
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