高效降胆固醇降甘油三酯乳酸菌的分离鉴定和功能分析
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摘要
筛选出高效降胆固醇降甘油三酯(Triglyceride,TG)菌株,为新型药物、保健食品的研制提供优良菌株。从6种西北特有样品中筛选分离出乳酸菌,采用磷硫铁比色法测定菌株降胆固醇能力,并测定菌株降甘油三酯能力,通过抑菌实验、耐酸、耐胆盐及小鼠体内毒性试验研究菌株各项性能,利用生理生化及16S rDNA法对菌株进行鉴定。结果表明,本试验筛选出3株高效降胆固醇降甘油三酯乳酸菌,其中菌株ZL010降胆固醇、降甘油三酯效果最强,蛋黄中胆固醇的降解率为73.65%,高纯度胆固醇降解率为40.06%,甘油三酯降解率为5.10%。3株菌都具有很好的抑菌、耐酸、耐胆盐能力,菌株经生理生化及16S rDNA鉴定ZL005为耐久肠球菌(Enterococcus durans),ZL010为植物乳杆菌(Lactobacillus plantarum),ZL016为戊糖乳杆菌(Lactobacillus pentosus)。小鼠毒性试验表明,菌株无毒副作用,小鼠各脏器一切正常,菌株可用于降脂降胆固醇保健药品及功能食品的制备。
In order to provide excellent strains for the preparation of new drugs and food,the cholesterol-lowering and triglyceride-lowering lactic acid bacteria were screened. Lactic acid bacteria were isolated from six samples of northwest endemic species. Triglyceride degradation and cholesterol-lowering abilities were determined by phosphorus-sulfur-iron colorimetry method.The bacteriostatic test,acid and bile salt tolerance test and toxicity test in mice were used to study the properties of the strain.The strain was identified by physiological and biochemical methods and 16 S rDNA.The results showed that there were 3 cholesterol-lowering and triglyceride-lowering lactic acid bacteria.The strain ZL010 had the strongest effect that the egg yolk cholesterol degradation rate was 73.65%,the high purity cholesterol degradation rate was 40.06%,the triglyceride degradation rate was 5.1%. The 3 strains had good bacteriostatic ability,and showed strong tolerance to acid and bile salt.The 16 S rDNA identified strain ZL005,ZL010,ZL016 as Enterococcus durans,Lactobacillus plantarum,Lactobacillus pentosus.The toxicity test revealed the strain has no toxic and side effects,the mice were dissected and observed that all the organs were normal. The strains could be used in the preparation of lipid-lowering and cholesterol-lowering drugs and functional foods.
In order to provide excellent strains for the preparation of new drugs and food,the cholesterol-lowering and triglyceride-lowering lactic acid bacteria were screened. Lactic acid bacteria were isolated from six samples of northwest endemic species. Triglyceride degradation and cholesterol-lowering abilities were determined by phosphorus-sulfur-iron colorimetry method.The bacteriostatic test,acid and bile salt tolerance test and toxicity test in mice were used to study the properties of the strain.The strain was identified by physiological and biochemical methods and 16 S rDNA.The results showed that there were 3 cholesterol-lowering and triglyceride-lowering lactic acid bacteria.The strain ZL010 had the strongest effect that the egg yolk cholesterol degradation rate was 73.65%,the high purity cholesterol degradation rate was 40.06%,the triglyceride degradation rate was 5.1%. The 3 strains had good bacteriostatic ability,and showed strong tolerance to acid and bile salt.The 16 S rDNA identified strain ZL005,ZL010,ZL016 as Enterococcus durans,Lactobacillus plantarum,Lactobacillus pentosus.The toxicity test revealed the strain has no toxic and side effects,the mice were dissected and observed that all the organs were normal. The strains could be used in the preparation of lipid-lowering and cholesterol-lowering drugs and functional foods.
引文
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[23]杜仁鹏,赵丹,王晓宇,等.1株乳酸高产菌的分离鉴定与系统发育分析[J].中国食品学报,2017,9(17):243-250.
[24]Qian C,Liu N,Yan X,et al.Engineering a highperformance,metagenomic-derived novel xylanase with improved soluble protein yield and thermostability[J].Enzyme and Microbial Technology,2015,70:35-41.
[2]于志会.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[D].长春:吉林大学,2013:17-19.
[3]陈伟伟,王文,隋辉,等.《中国心血管病报告2016》要点解读[J].中华高血压杂志,2017,25(7):605-608.
[4]吕秀红,陈凯飞,朱祺,等.降胆固醇乳酸菌的筛选与鉴定[J].中国食品学报,2016,16(3):198-204.
[5]Anila K,Kunzes A,Bhalla T C.In vitro cholesterol lassimilation and functional enzymatic activities of putative probiotic Lactobacillus sp.solated from fermented foods/beverages of North West India[J].Nutrition and Food Sciences,2016,6(2):2-5.
[6]Ziaeian B,Dinkler J,Watson K.Implementation of the 2013American college of cardiology/American heart association blood cholesterol guideline including data from the improved reduction of outcomes:Vytorin efficacy international trial[J].Reviews in Cardiovascular Medicine,2015,16(2):125-130.
[7]Anderson F,Mbatha S Z,Thomson S R.The early management of pancreatitis associated with hyper-triglyceridaemia[J].South African Journal of Surgery,2011,49(2):82-84.
[8]谌素华,王维民,蔡璐,等.马尾藻岩藻聚糖硫酸酯对高脂血症小鼠脂质代谢相关酶的影响[J].中国食品学报,2017,17(2):10-16.
[9]Mancini G B,Baker S,Bergeron J,et al.Diagnosis,prevention,and management of statin adverse effects and intolerance:Canadian consensus working group update(2016)[J].The Canadian Journal of Cardiology,2016,32(7):35-65.
[10]Edel A L,Rodriguezleyva D,Maddaford T G,et al.Dietary flaxseed independently lowers circulating cholesterol and lowers it beyond the effects of cholesterol-lowering medications alone inpatients with peripheral artery disease[J].Journal of Nutrition,2015,145(4):749-757.
[11]Barone E,Di D F,Butterfield D A.Statins more than cholesterol lowering agents in Alzheimer disease:Their pleiotropic functions as potential therapeutic targets[J].Biochemical Pharmacology,2014,88(4):605-616.
[12]Tomaszewski M,StpieńKM,Tomaszewska J,et al.Statininduced myopathies[J].Pharmacological Reports,2011,63(4):859-866.
[13]袁成莹,候原平,常晶,等.他汀类药物在临床应用中的主要不良反应及防治策略[J].中华老年多器官疾病杂志2018,17(12):949-951.
[14]梁茂本.汀类药物不良反应87例文献分析[J].中国生化药物杂志,2009,30(5):336-338.
[15]陈珂可,朱建国,姜甜,等.具有降解胆固醇能力的益生菌菌株筛选[J].食品科技,2018,43(11):7-11.
[16]王辑,顾芸佳,马文慧,等.内蒙古奶豆腐中潜在益生性乳酸菌的筛选[J].食品科学,2014,35(13):171-177.
[17]Li Jinjie,Zhang Wen,Wang Chuan,et al.Lactococcus lactis expressing food-gradeβ-galactosidase alleviates lactose intolerance symptoms in post-weaning BALB/c mice[J].Applied Microbiology and Biotechnology,2012,96(6):1499-1506.
[18]Choi E A,Chang H C.Cholesterol-lowering effects of a putative probiotics in Lactobacillus plantarum EM isolated from kimchi[J].Food Science and Technology,2015,62(1):210-217.
[19]吴慧昊,段龙飞,张园园,等.熊猫粪便中乳酸菌的分离鉴定与降胆固醇作用研究[J].黑龙江畜牧兽医,2011,6(383):14-17.
[20]臧荣鑫,杨具田.生物化学实验教程[M].兰州:兰州大学出版社,2010:159-161.
[21]佘之蕴,黄宝莹,刘海卿,等.牛津杯法测定食品添加剂对五种益生菌的抑菌活力[J].食品工业,2016(1):171-174.
[22]Jeun J,Kim S,Cho S,et al.Hypocholesterolemic effects of Lactobacillus plantarum KCTC3928 by increased bile acid excretion in C57BL/6 mice[J].Nutrition,2010,26(3):321-330.
[23]杜仁鹏,赵丹,王晓宇,等.1株乳酸高产菌的分离鉴定与系统发育分析[J].中国食品学报,2017,9(17):243-250.
[24]Qian C,Liu N,Yan X,et al.Engineering a highperformance,metagenomic-derived novel xylanase with improved soluble protein yield and thermostability[J].Enzyme and Microbial Technology,2015,70:35-41.