西藏曲拉和发酵乳中抗氧化和益生特性乳酸菌的筛选及鉴定
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摘要
以抗过氧化氢、清除自由基为抗氧化指标,耐酸、耐胆盐为益生特性指标,对采集自西藏的自然发酵曲拉和发酵乳中乳酸菌进行了分离、筛选和鉴定。结果表明:从9份样品中分得的62株菌有23株对0.3%过氧化氢具备耐受性,其中4-C、5-D、5-E、9-E对DPPH·和·OH清除率最高分别为86.97%±0.11%、27.62%±0.28%;经鉴定5-D、5-E、9-E、4-C分别为Lactobacillus delbrueckii subsp. Jakobsenii、Lactobacillus delbrueckii subsp. Jakobsenii、Lactobacillus gasseri、Pediococcus pentosaceus;此外,菌株5-D、5-E对酸碱和0.3%胆盐均有较好的耐受能力,具备抗氧化和益生特性,有较大潜力应用于天然抗氧化剂的研究与开发,同时也丰富了西藏地区传统牦牛乳制品中乳酸菌资源库的挖掘工作。
This study isolated and identified lactic acid bacteria from the fermented milk and Qula in Tibet,through antioxidant indicators of anti-hydrogen peroxide and scavenging free radicals and probiotic properties of resistance to acid and bile salt.The results showed that 62 strains of bacteria were acquired,and 23 strains of them were tolerant to 0.3% hydrogen peroxide.Among the 23 strains,4-C,5-D,5-E and 9-E had the highest scavenging capacity of DPPH· and ·OH,with the rate of 86.97% ±0.11% and 27.62% ± 0.28% respectively.The strains of 5-D,5-E,9-E and 4-C were identified as Lactobacillus delbrueckii subsp. Jakobsenii,Lactobacillus delbrueckii subsp. Jakobsenii,Lactobacillus gasseri,Pediococcus pentosaceus respectively. In addition,the 5-D and 5-E had the capability of concurrently antioxidant and probiotic properties of high tolerance to acidbase and 0.3% bile salt stress.They had enormous potential in the research and development of natural antioxidants,as well as enriched the excavation of lactic acid bacteria resources of traditional yak dairy products in Tibet area.
This study isolated and identified lactic acid bacteria from the fermented milk and Qula in Tibet,through antioxidant indicators of anti-hydrogen peroxide and scavenging free radicals and probiotic properties of resistance to acid and bile salt.The results showed that 62 strains of bacteria were acquired,and 23 strains of them were tolerant to 0.3% hydrogen peroxide.Among the 23 strains,4-C,5-D,5-E and 9-E had the highest scavenging capacity of DPPH· and ·OH,with the rate of 86.97% ±0.11% and 27.62% ± 0.28% respectively.The strains of 5-D,5-E,9-E and 4-C were identified as Lactobacillus delbrueckii subsp. Jakobsenii,Lactobacillus delbrueckii subsp. Jakobsenii,Lactobacillus gasseri,Pediococcus pentosaceus respectively. In addition,the 5-D and 5-E had the capability of concurrently antioxidant and probiotic properties of high tolerance to acidbase and 0.3% bile salt stress.They had enormous potential in the research and development of natural antioxidants,as well as enriched the excavation of lactic acid bacteria resources of traditional yak dairy products in Tibet area.
引文
[1]蒋厚阳,陈芝兰,赵国华,等.PCR-DGGE法分析西藏传统发酵乳制品中乳酸菌的多样性[J].食品科学,2014,35(1):167-173.
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[3]王军,王琳琳,韩玲,等.曲拉热风干燥工艺参数筛选及色泽品质评价[J].农业工程学报,2014,30(16):296-304.
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[7]罗章,陈历水,陈历俊,等.藏灵菇乳中具有优良抗氧化活性乳酸菌的筛选鉴定[J].食品科学,2015,36(3):109-113.
[8]骞宇,赵欣,李银聪,等.青藏高原自然发酵牦牛酸奶中乳酸菌的抗氧化能力的研究[J].食品工业科技,2014,35(3):119-122.
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[13]罗红玉,郁军,岳鹏翔,等.茶渣多肽制备及其对羟自由基的清除能力[J].食品科学,2011,32(14):61-66.
[14]陈芝兰,程池,马凯,等.西藏地区牦牛发酵乳制品中乳酸菌的分离与鉴定[J].食品科学,2008,29(12):408-412.
[15]李利,孔丽,张宁,等.耐酸耐胆盐乳酸菌的鉴定及筛选[J].食品科学,2015,36(21):123-128.
[16]刘宏宇,汪立平,艾连中,等.乳酸菌的抗氧化活性和耐酸耐胆盐性能的研究[J].食品工业科技,2014,35(2):92-96.
[17]Marteau P,Minekus M,Havenaar R,et al.Survival of lactic acid bacteria in a dynamic model of the stomach and small intestine:validation and the effects of bile[J]. Journal of Dairy Science,1997,80(6):1031-1037.
[18]赵芳,李艳琴,李彬春.模拟人体胃肠道环境筛选益生乳杆菌[J].微生物学通报,2016,43(6):1396-1403.
[19]张凤敏,田丰伟,陈卫,等.具抗氧化活性乳酸菌的筛选[J].中国乳品工业,2007,35(2):4-7.
[20]韦献雅,殷丽琴,钟成,等.DPPH法评价抗氧化活性研究进展[J].食品科学,2014,35(9):317-322.
[21]文镜,张西,陈曦,等.体内羟自由基的检测方法[J].食品科学,2004,25(10):351-357.
[22]杨郁荭,白明.乳酸菌抗氧化机理的初步探讨[J].中国乳业,2011(7):68-73.
[23]刘珊春,赵欣,李键,等.高抗氧化乳酸菌的筛选鉴定[J].食品与发酵工业,2017,43(8):59-66.
[24]Adimpong D B,Nielsen D S,Srensen K I,et al.Lactobacillus delbrueckii subsp.jakobsenii subsp.nov. isolated from dolo wort,an alcoholic fermented beverage in Burkina Faso[J]. International Journal of Systematic and Evolutionary Microbiology,2013,63(10):3720-3726.
[25]宋宇琴,孙志宏,张和平.德氏乳杆菌保加利亚亚种遗传多样性和种群结构的MLST分析免疫原性死亡的作用研究[C]//乳酸菌与健康国际研讨会,2015.
[26]赵小茜,魏旭丹,陈戴玲,等.乳酸菌耐酸耐胆盐机制研究进展[J].乳业科学与技术,2017,40(3):33-36.
[27]胡爱华,敖晓琳,陈岑,等.乳酸菌耐酸耐胆盐机制的研究进展[J].食品工业科技,2015,36(8):380-383.
[2]Ren Y,Yang Y,Zhang D,et al. Diversity analysis and quantification of lactic acid bacteria in traditionally fermented yaksTMmilk products from Tibet[J]. Food Biotechnology,2017,31(1):1-19.
[3]王军,王琳琳,韩玲,等.曲拉热风干燥工艺参数筛选及色泽品质评价[J].农业工程学报,2014,30(16):296-304.
[4]Alexandra S,Joachim J,JoséP C,et al. Oxidative stress in chronic kidney disease[J]. Oxidative Medicine and Cellular Longevity,2016,2016(3):165-179.
[5]Syburra C,Passi S.Oxidative stress in patients with multiple sclerosis[J]. Ukrainskyi Biokhimichnyi Zhurnal,2016,71(3):112.
[6]张卓睿,毛迪锐,高晗,等.蓝莓花青素对小鼠抗疲劳及体内抗氧化作用[J].食品科学,2017,38(21):207-211.
[7]罗章,陈历水,陈历俊,等.藏灵菇乳中具有优良抗氧化活性乳酸菌的筛选鉴定[J].食品科学,2015,36(3):109-113.
[8]骞宇,赵欣,李银聪,等.青藏高原自然发酵牦牛酸奶中乳酸菌的抗氧化能力的研究[J].食品工业科技,2014,35(3):119-122.
[9]陈明,柯文灿,保安安,等.青藏高原牦牛酸奶中具高抗氧化能力乳酸菌的筛选[J].食品工业科技,2016,37(8):201-205.
[10]杨洁彬.乳酸菌:生物学基础及应用[M].北京:中国轻工业出版社,1996.
[11]王刚,田丰伟,刘小鸣,等.2株具有优良体外抗氧化能力乳酸菌的筛选与鉴定[J].食品工业科技,2013,34(15):149-153.
[12]Lin M Y,Chang F J.Antioxidative effect of intestinal bacteria Bifidobacterium longum ATCC 15708 and Lactobacillus acidophilus ATCC 4356.[J]. Digestive Diseases&Sciences,2000,45(8):1617-1622.
[13]罗红玉,郁军,岳鹏翔,等.茶渣多肽制备及其对羟自由基的清除能力[J].食品科学,2011,32(14):61-66.
[14]陈芝兰,程池,马凯,等.西藏地区牦牛发酵乳制品中乳酸菌的分离与鉴定[J].食品科学,2008,29(12):408-412.
[15]李利,孔丽,张宁,等.耐酸耐胆盐乳酸菌的鉴定及筛选[J].食品科学,2015,36(21):123-128.
[16]刘宏宇,汪立平,艾连中,等.乳酸菌的抗氧化活性和耐酸耐胆盐性能的研究[J].食品工业科技,2014,35(2):92-96.
[17]Marteau P,Minekus M,Havenaar R,et al.Survival of lactic acid bacteria in a dynamic model of the stomach and small intestine:validation and the effects of bile[J]. Journal of Dairy Science,1997,80(6):1031-1037.
[18]赵芳,李艳琴,李彬春.模拟人体胃肠道环境筛选益生乳杆菌[J].微生物学通报,2016,43(6):1396-1403.
[19]张凤敏,田丰伟,陈卫,等.具抗氧化活性乳酸菌的筛选[J].中国乳品工业,2007,35(2):4-7.
[20]韦献雅,殷丽琴,钟成,等.DPPH法评价抗氧化活性研究进展[J].食品科学,2014,35(9):317-322.
[21]文镜,张西,陈曦,等.体内羟自由基的检测方法[J].食品科学,2004,25(10):351-357.
[22]杨郁荭,白明.乳酸菌抗氧化机理的初步探讨[J].中国乳业,2011(7):68-73.
[23]刘珊春,赵欣,李键,等.高抗氧化乳酸菌的筛选鉴定[J].食品与发酵工业,2017,43(8):59-66.
[24]Adimpong D B,Nielsen D S,Srensen K I,et al.Lactobacillus delbrueckii subsp.jakobsenii subsp.nov. isolated from dolo wort,an alcoholic fermented beverage in Burkina Faso[J]. International Journal of Systematic and Evolutionary Microbiology,2013,63(10):3720-3726.
[25]宋宇琴,孙志宏,张和平.德氏乳杆菌保加利亚亚种遗传多样性和种群结构的MLST分析免疫原性死亡的作用研究[C]//乳酸菌与健康国际研讨会,2015.
[26]赵小茜,魏旭丹,陈戴玲,等.乳酸菌耐酸耐胆盐机制研究进展[J].乳业科学与技术,2017,40(3):33-36.
[27]胡爱华,敖晓琳,陈岑,等.乳酸菌耐酸耐胆盐机制的研究进展[J].食品工业科技,2015,36(8):380-383.