益生性降胆固醇植物乳杆菌的筛选、发酵特性及体内功效研究
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摘要
益生性降胆固醇植物乳杆菌的筛选、发酵特性及体内功效研究内容隶属于农业部现代农业产业技术体系建设专项资金资助项目（项目编号：cyrs-37）。
血清胆固醇水平偏高是诱发动脉硬化、冠心病、脑中风等心脑血管疾病的主要危险因素，严重威胁着人类的健康。因此，保持人体内胆固醇含量在正常范围内对于预防心脑血管疾病是至关重要的。本文从传统发酵食品中筛选出11株具有降胆固醇功能的植物乳杆菌，对其益生特性、体外降胆固醇机理进行研究。从中筛选1株植物乳杆菌为优选菌株，对其进行发酵特性的研究，并研制功能性酸奶。在此基础上，利用动物实验对所选菌株发酵乳的降血脂功效及抗氧化作用进行了测定。初步得出如下结论：
⑴以分离自东北家庭自制酸菜及内蒙古奶豆腐中的11株植物乳杆菌为研究对象，采用邻苯二甲醛法筛选具有降胆固醇功能的植物乳杆菌，并通过耐酸、耐胆盐、抗生素敏感性、抑菌能力、粘附能力及β-乳糖酶活性实验，分析实验菌株的益生特性。从中筛选获得了4株（S2-5，S2-6，S4-1、C88）具有较强的耐酸、耐胆盐等益生特性和降胆固醇功效的植物乳杆菌，并最终确定Lactobacillus plantarum C88菌株为筛选目标菌株。
⑵进一步对植物乳杆菌S2-5，S2-6，S4-1、C88的体外降胆固醇作用研究发现，实验菌株主要是通过同化吸收与共沉淀两种方式共同降低培养基中的胆固醇。
⑶对Lactobacillus plantarum C88发酵特性研究表明：Lactobacillus plantarum C88的凝乳活性较弱，蛋白水解活性较高；4℃贮28d，该菌株表现出良好的存活能力，活菌数高于L. rhamnosus GG。添加了菊粉、葡萄糖和低聚果糖等，可以促进Lactobacillus plantarumC88的活菌数目提高。
⑷由于Lactobacillus plantarum C88具有良好的功能特性，为了更好地发挥其益生作用，将其应用于产品中具有更重要的现实意义。因此，通过对其产酸能力实验、拮抗实验以及与商业酸奶发酵剂YO-MIX复配制备酸奶实验，结果表明：Lactobacillus plantarum C88对商业发酵剂菌株没有抑制作用，在乳中的存活能力较强，对酸奶的pH、滴定酸度、凝胶强度以及持水力均无不良影响，可提高酸奶的粘度，改善酸奶的感官特性。
⑸为了了解筛选具有高降胆固醇及益生特性菌株发酵乳在动物体内的降胆固醇功效，参照保健食品功能性评价方法和程序，将Lactobacillus plantarum C88、Lactobacillusplantarum S4-1发酵乳灌喂高血脂模型小鼠。结果表明：Lactobacillus plantarum C88、L.plantarum S4-1高、低剂量发酵乳显著降低了小鼠血清总胆固醇（TC）、低密度胆固醇（LDL-C）、甘油三酯（TG）的水平；L. plantarum C88高剂量发酵乳能显著提高血清中HDL-C的含量；L. plantarum C88发酵乳体内降胆固醇的效果强于L. plantarum S4-1发酵乳。
⑹采用Lactobacillus plantarum C88发酵乳饲喂由D-半乳糖连续皮下注射诱导的亚急性衰老模型小鼠，结果表明：Lactobacillus plantarum C88发酵乳能显著地提高衰老模型小鼠血清和肝组织中SOD活性、肝组织中GSH-Px活性及T-AOC，同时降低小鼠血清和肝组织中MDA含量。Lactobacillus plantarum C88发酵乳对衰老模型小鼠有一定的抗氧化作用。
Screening、Fermentation properties and in vivo Efficacy of cholesterol-lowering ProbioticLactobacillus plantarum Strains is Supported by Department of agriculture modern agricultureindustry technology system construction of special funds of funded projects (No. cyrs-37).
Overmuch cholesterol in the blood and diet has been recognized as a major risk factor forcoronary heart diseases and colon cancer, and many health conscious people are adoptingmeasures to decrease risk by reducing blood serum cholesteorl. A total of11Lactobacillusplantarum isolated from traditionally fermented food with cholesterol-reducing and probioticproperties were tested. One strain was selected as the optimal strain for all its tested abilities. Thefermentation characteristics and functional yoghurt of the strain was studied. The reduce serumcholesterol and antioxidant effects of selected strains of fermented milk were measured. Thesegives useful information for further exploitation and utilization of lactic acid bacteria intraditional fermented milk products．
⑴Eleven L. plantarum strains isolated from the Northeast homemade sauerkraut and milktofo in Mongolia were screened for cholesterol-lowering functional by using σ-phthalaldehyde.By acid and bile tolerance and antimicrobial susceptibility testing, these strains were studied fortheir potentic probiotic.4strains (S2-5、S2-6、S4-1、C88)were screened for their excellentcholesterol-reducing ability and probiotic properties. Strain C88was selected as the optimalstrain for all its tested abilities.
⑵More studies on the mechanism of cholesterol removal in vitro was carried out. Theresults showed that strains S2-5、S2-6、S4-1、C88were able to precipitate and assimilate cholesterols.
⑶The fermentation properties of L. plantarum C88was assayed. The results showed that: L.plantarum C88displayed weak milk-coagulating activity, but strong proteolytic activity; L.plantarum C88maintained high viability in fermented milk during storage compared with L.rhamnosus GG. Added probiotics (inulin, glucose and fructooligosaccharides etc) can increasethe number of viable cells of L. plantarum C88.
⑷Due to the functionality of L. plantarum C88, it will have a important meaning that isapplied to milk industry. Through the milk acidification, antagonistic activity and mix withcommercial yogurt culture YO-MIX was tested by yogurt experiment. The results showed that L.plantarum C88displayed low milk acidification and high survival ability, while no inhibitionwas observed towards the commercial yogurt starters used. Yogurt produced with L. plantarumC88exhibited no effect to the yogurt pH, titratable acidity, gel strength, water holding capacity.The strain not only increases the viscosity of yogurt but also promotes the sensory index ofyogurt.
⑸T he effects ofL. plantarum C88、L. plantarum S4-1on serum lipids were estimated inICR mice fed cholesterol-enriched diets. The results showed that L. plantarum C88、L.plantarum S4-1feeding significantly lowered the serum total cholesterol, low-density lipoproteincholesterol and triglycerides level, meantime L. plantarum C88improved the content ofhigh-density lipoprotein cholesterol.
⑹ICR subacute aged model mice, which were given subcutaneous injection of D-galactosecontinuously, were fed with fermented milk containing L. plantarum C88. The results showedthat L. plantarum C88fermented milk could significantly enhance the aged mice serum and livertissue SOD activity, liver tissue GSH-Px activity and liver tissue total antioxidant capacity(T-AOC), lowered the serum and liver tissue MDA content. It proved that the L.plantarum C88fermented milk has certain antioxidant effects on aged mice.
血清胆固醇水平偏高是诱发动脉硬化、冠心病、脑中风等心脑血管疾病的主要危险因素，严重威胁着人类的健康。因此，保持人体内胆固醇含量在正常范围内对于预防心脑血管疾病是至关重要的。本文从传统发酵食品中筛选出11株具有降胆固醇功能的植物乳杆菌，对其益生特性、体外降胆固醇机理进行研究。从中筛选1株植物乳杆菌为优选菌株，对其进行发酵特性的研究，并研制功能性酸奶。在此基础上，利用动物实验对所选菌株发酵乳的降血脂功效及抗氧化作用进行了测定。初步得出如下结论：
⑴以分离自东北家庭自制酸菜及内蒙古奶豆腐中的11株植物乳杆菌为研究对象，采用邻苯二甲醛法筛选具有降胆固醇功能的植物乳杆菌，并通过耐酸、耐胆盐、抗生素敏感性、抑菌能力、粘附能力及β-乳糖酶活性实验，分析实验菌株的益生特性。从中筛选获得了4株（S2-5，S2-6，S4-1、C88）具有较强的耐酸、耐胆盐等益生特性和降胆固醇功效的植物乳杆菌，并最终确定Lactobacillus plantarum C88菌株为筛选目标菌株。
⑵进一步对植物乳杆菌S2-5，S2-6，S4-1、C88的体外降胆固醇作用研究发现，实验菌株主要是通过同化吸收与共沉淀两种方式共同降低培养基中的胆固醇。
⑶对Lactobacillus plantarum C88发酵特性研究表明：Lactobacillus plantarum C88的凝乳活性较弱，蛋白水解活性较高；4℃贮28d，该菌株表现出良好的存活能力，活菌数高于L. rhamnosus GG。添加了菊粉、葡萄糖和低聚果糖等，可以促进Lactobacillus plantarumC88的活菌数目提高。
⑷由于Lactobacillus plantarum C88具有良好的功能特性，为了更好地发挥其益生作用，将其应用于产品中具有更重要的现实意义。因此，通过对其产酸能力实验、拮抗实验以及与商业酸奶发酵剂YO-MIX复配制备酸奶实验，结果表明：Lactobacillus plantarum C88对商业发酵剂菌株没有抑制作用，在乳中的存活能力较强，对酸奶的pH、滴定酸度、凝胶强度以及持水力均无不良影响，可提高酸奶的粘度，改善酸奶的感官特性。
⑸为了了解筛选具有高降胆固醇及益生特性菌株发酵乳在动物体内的降胆固醇功效，参照保健食品功能性评价方法和程序，将Lactobacillus plantarum C88、Lactobacillusplantarum S4-1发酵乳灌喂高血脂模型小鼠。结果表明：Lactobacillus plantarum C88、L.plantarum S4-1高、低剂量发酵乳显著降低了小鼠血清总胆固醇（TC）、低密度胆固醇（LDL-C）、甘油三酯（TG）的水平；L. plantarum C88高剂量发酵乳能显著提高血清中HDL-C的含量；L. plantarum C88发酵乳体内降胆固醇的效果强于L. plantarum S4-1发酵乳。
⑹采用Lactobacillus plantarum C88发酵乳饲喂由D-半乳糖连续皮下注射诱导的亚急性衰老模型小鼠，结果表明：Lactobacillus plantarum C88发酵乳能显著地提高衰老模型小鼠血清和肝组织中SOD活性、肝组织中GSH-Px活性及T-AOC，同时降低小鼠血清和肝组织中MDA含量。Lactobacillus plantarum C88发酵乳对衰老模型小鼠有一定的抗氧化作用。
Screening、Fermentation properties and in vivo Efficacy of cholesterol-lowering ProbioticLactobacillus plantarum Strains is Supported by Department of agriculture modern agricultureindustry technology system construction of special funds of funded projects (No. cyrs-37).
Overmuch cholesterol in the blood and diet has been recognized as a major risk factor forcoronary heart diseases and colon cancer, and many health conscious people are adoptingmeasures to decrease risk by reducing blood serum cholesteorl. A total of11Lactobacillusplantarum isolated from traditionally fermented food with cholesterol-reducing and probioticproperties were tested. One strain was selected as the optimal strain for all its tested abilities. Thefermentation characteristics and functional yoghurt of the strain was studied. The reduce serumcholesterol and antioxidant effects of selected strains of fermented milk were measured. Thesegives useful information for further exploitation and utilization of lactic acid bacteria intraditional fermented milk products．
⑴Eleven L. plantarum strains isolated from the Northeast homemade sauerkraut and milktofo in Mongolia were screened for cholesterol-lowering functional by using σ-phthalaldehyde.By acid and bile tolerance and antimicrobial susceptibility testing, these strains were studied fortheir potentic probiotic.4strains (S2-5、S2-6、S4-1、C88)were screened for their excellentcholesterol-reducing ability and probiotic properties. Strain C88was selected as the optimalstrain for all its tested abilities.
⑵More studies on the mechanism of cholesterol removal in vitro was carried out. Theresults showed that strains S2-5、S2-6、S4-1、C88were able to precipitate and assimilate cholesterols.
⑶The fermentation properties of L. plantarum C88was assayed. The results showed that: L.plantarum C88displayed weak milk-coagulating activity, but strong proteolytic activity; L.plantarum C88maintained high viability in fermented milk during storage compared with L.rhamnosus GG. Added probiotics (inulin, glucose and fructooligosaccharides etc) can increasethe number of viable cells of L. plantarum C88.
⑷Due to the functionality of L. plantarum C88, it will have a important meaning that isapplied to milk industry. Through the milk acidification, antagonistic activity and mix withcommercial yogurt culture YO-MIX was tested by yogurt experiment. The results showed that L.plantarum C88displayed low milk acidification and high survival ability, while no inhibitionwas observed towards the commercial yogurt starters used. Yogurt produced with L. plantarumC88exhibited no effect to the yogurt pH, titratable acidity, gel strength, water holding capacity.The strain not only increases the viscosity of yogurt but also promotes the sensory index ofyogurt.
⑸T he effects ofL. plantarum C88、L. plantarum S4-1on serum lipids were estimated inICR mice fed cholesterol-enriched diets. The results showed that L. plantarum C88、L.plantarum S4-1feeding significantly lowered the serum total cholesterol, low-density lipoproteincholesterol and triglycerides level, meantime L. plantarum C88improved the content ofhigh-density lipoprotein cholesterol.
⑹ICR subacute aged model mice, which were given subcutaneous injection of D-galactosecontinuously, were fed with fermented milk containing L. plantarum C88. The results showedthat L. plantarum C88fermented milk could significantly enhance the aged mice serum and livertissue SOD activity, liver tissue GSH-Px activity and liver tissue total antioxidant capacity(T-AOC), lowered the serum and liver tissue MDA content. It proved that the L.plantarum C88fermented milk has certain antioxidant effects on aged mice.
引文
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