内蒙古科尔沁地区食疗用酸马奶发酵引子化学成分及细菌多样性分析
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摘要
对内蒙古科尔沁地区食疗用酸马奶发酵引子部分化学指标,包括pH值、糖类、有机酸、游离脂肪酸等进行分析,同时为调查其中细菌多样性,应用Illumina MiSeq第2代测序技术测定其细菌的16S rDNA V3-V4高变区序列,分析物种的丰度、物种分布和Alpha多样性。结果:该酸马奶引子样品pH值为3.54,乳糖、半乳糖质量分数分别为(0.281±0.011)%和(0.100±0.003)%,未检测到葡萄糖;乳酸质量分数为(1.506±0.069)%,丙酸质量分数为(0.053±0.002)%,乙酸质量分数为(0.345±0.014)%,丁酸质量分数为(1.143±0.061)%;共检测到19种脂肪酸,包括12种饱和脂肪酸和7种不饱和脂肪酸;不饱和脂肪酸中α-亚麻酸质量浓度最高,为(14.12±0.36)mg/L;细菌多样性检测结果表明,该样品所获优化序列为(62 082.7±4 868.8)条,可操作分类单元(operational taxonomic units,OTU)数为34.3±2.3;稀释曲线表明该测序深度充分,OTU数接近于饱和。该酸马奶引子细菌主要分布于以下4个属:乳杆菌属(Lactobacillus)84.08%、醋杆菌属(Acetobacter)9.83%、乳球菌属(Lactococcus)2.41%、链球菌属(Streptococcus)2.18%;其优势菌属为乳杆菌属。以上结果表明,该酸马奶引子不饱和脂肪酸含量丰富,益生菌占主要地位,食品安全有保障。
In this study, some chemical properties including pH, carbohydrates, organic acids and free fatty acids of a fermentation starter for dietotherapeutic koumiss in Kerqin, Inner Mongolia were analyzed. To investigate the diversity of the bacterial community in the starter culture, second-generation sequencing of the V3-V4 region of the 16 S rDNA genes was performed on the Illumina MiSeq platform. Thereafter, the species abundance, distribution and alpha diversity were analyzed. The analytical results indicated that the pH averaged at 3.54; the mean values of lactose and galactose were(0.281 ± 0.011)% and(0.100 ± 0.003)% respectively, and no glucose was detected. The levels of lactic acid, propanoic acid,acetic acid and butyric acid were(1.506 ± 0.069)%,(0.053 ± 0.002)%,(0.345 ± 0.014)%, and(1.143 ± 0.061)%, respectively.A total of 19 fatty acids were detected, including 12 saturated fatty acids and 7 unsaturated fatty acids. α-Linolenic acid was the most abundant unsaturated fatty acid at a level of(14.12 ± 0.36) mg/L. The results of bacterial diversity detection showed that(62 082.7 ± 4 868.8) clean reads were obtained, and the number of operational taxonomic units(OTUs) was 34.3 ± 2.3.The rarefaction curve showed that the sequencing depth was sufficient and the number of OTUs was close to saturation. The bacteria community in this sample was mainly distributed in the following 4 genera: Lactobacillus(84.08%), Acetobacter(9.83%), Lactobacillus(2.41%), and Streptococcus(2.18%) with the dominant genus being Lactobacillus. The above results showed that this fermentation starter sample was rich in unsaturated fatty acids and probiotics, and could guarantee food safety.
In this study, some chemical properties including pH, carbohydrates, organic acids and free fatty acids of a fermentation starter for dietotherapeutic koumiss in Kerqin, Inner Mongolia were analyzed. To investigate the diversity of the bacterial community in the starter culture, second-generation sequencing of the V3-V4 region of the 16 S rDNA genes was performed on the Illumina MiSeq platform. Thereafter, the species abundance, distribution and alpha diversity were analyzed. The analytical results indicated that the pH averaged at 3.54; the mean values of lactose and galactose were(0.281 ± 0.011)% and(0.100 ± 0.003)% respectively, and no glucose was detected. The levels of lactic acid, propanoic acid,acetic acid and butyric acid were(1.506 ± 0.069)%,(0.053 ± 0.002)%,(0.345 ± 0.014)%, and(1.143 ± 0.061)%, respectively.A total of 19 fatty acids were detected, including 12 saturated fatty acids and 7 unsaturated fatty acids. α-Linolenic acid was the most abundant unsaturated fatty acid at a level of(14.12 ± 0.36) mg/L. The results of bacterial diversity detection showed that(62 082.7 ± 4 868.8) clean reads were obtained, and the number of operational taxonomic units(OTUs) was 34.3 ± 2.3.The rarefaction curve showed that the sequencing depth was sufficient and the number of OTUs was close to saturation. The bacteria community in this sample was mainly distributed in the following 4 genera: Lactobacillus(84.08%), Acetobacter(9.83%), Lactobacillus(2.41%), and Streptococcus(2.18%) with the dominant genus being Lactobacillus. The above results showed that this fermentation starter sample was rich in unsaturated fatty acids and probiotics, and could guarantee food safety.
引文
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[10]田颖刚,谢明勇,付志红,等.乌骨鸡脂肪油中脂肪酸组成的气相色谱-质谱分析[J].南昌大学学报(理科版),2006,30(3):264-266.
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[13]额尔登孟克,潘琳,阿木尔图布兴,等.传统发酵乳制品酸马奶的研究进展[J].内蒙古农业大学学报(自然科学版),2016,37(2):134-139.
[14]拉·巴勒道尔吉,策·纳木斯来.蒙古艾日嘎[M].呼和浩特:内蒙古人民出版社,1990:137-138.
[15]布仁其其格,高雅罕,包艳青.酸马奶发酵过程中理化指标与细菌相的动态变化[J].食品工业科技,2016,37(11):118-121.
[16]ISHII S.Study on the kumiss(Airag)of Mongolian nomads after severe cold in the winters of 2000 and 2001[J].Milk Science,2003,52:49-52.
[17]郭本恒.功能性乳制品[M].北京:中国轻工业出版社,2001.
[18]ZHOU Y M,LIN J C,WU L P,et al.Effect of antecolic or retrocolic reconstruction of the gastro/duodenojejunostomy on delayed gastric emptying after pancreaticoduodenectomy:a meta-analysis[J].BMCGastroenterology,2015,15:68.DOI:10.1186/s12876-015-0300-8.
[19]QUIGLEY E M.Other forms of gastroparesis:postsurgical,parkinson,other neurologic diseases,connective tissue disorders[J].Gastroenterology Clinics of North America,2015,44(1):69-81.DOI:10.1016/j.gtc.2014.11.006.
[20]LACY B E.Functional dyspepsia and gastroparesis:one disease or two[J].American Journal of Gastroenterology,2012,107(11):1615-1620.DOI:10.1038/ajg.2012.104.
[21]PASRICHA P J.Future directions in the treatment of gastroparesis[J].Gastroenterology Clinics of North America,2015,44(1):185-189.DOI:10.1016/j.gtc.2014.11.014.
[22]SMITS M J,LOOTS C M,BENNINGA M A,et al.New insights in gastroesophageal reflux,esophageal function and gastric emptying in relation to dysphagia before and after anti-reflux surgery in children[J].Current Gastroenterology Reports,2013,15(10):351.DOI:10.1007/s11894-013-0351-3.
[23]GRASSI M,PETRACCIA L,MENNUNI G,et al.Changes,functional disorders,and diseases in the gastrointestinal tract of elderly[J].Nutricion Hospitalaria,2011,26(4):659-668.DOI:10.1590/S0212-16112011000400001.
[24]叶博.短链脂肪酸促进人和小鼠gastroids中产ghrelin细胞分化和小鼠胃排空的生理机制研究[D].上海:第二军医大学第一附属医院,2015.
[25]于昱,袁缨.多不饱和脂肪酸的营养研究[J].中国饲料,2003(24):21-23.DOI:10.3969/j.issn.1004-3314.2003.24.010.
[26]孙翔宇,高贵田,段爱莉,等.多不饱和脂肪酸的研究进展[J].食品工业科技,2012,33(7):418-423.DOI:10.13386/j.issn1002-0306.2012.07.030.
[27]孙天松,王俊国,张列兵,等.中国新疆地区酸马奶中乳酸菌生物多样性研究[J].微生物学通报,2007,34(3):451-454.DOI:10.3969/j.issn.0253-2654.2007.03.013.
[28]RING E,ANDERSEN R,SPERSTAD S,et al.Bacterial community of koumiss from Mongolia investigated by culture and cultureindependent methods[J].Food Biotechnology,2014,28(4):333-353.DOI:10.1080/08905436.2014.964253.
[29]Food and Agriculture Organization of United Nations,World Health Organization.Report on joint FAO/WHO expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria[S].Cordoba,Argentina,2001.
[30]李海燕,徐杰,云月英,等.传统发酵酸马奶中瑞士乳杆菌耐酸性的筛选[J].乳业科学与技术,2006,29(4):158-160.DOI:10.15922/j.cnki.jdst.2006.04.003.
[31]贾宏信,龚广予,郭本恒.益生菌干酪的研究进展[J].食品科学,2013,34(15):355-360.DOI:10.7506/spkx1002-6630-201315072.
[2]孙天松.中国新疆地区传统发酵酸马奶的化学组成及乳酸菌生物多样性[D].呼和浩特:内蒙古农业大学,2006.
[3]哈斯苏荣,阿木古楞,芒来.酸马奶及其医学价值[J].中国中药杂志,2003,28(1):11-14.DOI:10.3321/j.issn:1001-5302.2003.01.006.
[4]芒来.策格:草原珍品[M].呼和浩特:内蒙古人民出版社,2013:59.
[5]斯琴格日勒.蒙医酸马奶疗法辅助治疗作用体会[J].中国民族医药杂志,2012,18(2):21.DOI:10.3969/j.issn.1006-6810.2012.02.020.
[6]徐少泽.马奶、酸马奶的食疗和医用价值[J].中国民族医药杂志,2012,18(2):21-22.DOI:10.3969/j.issn.1006-6810.2012.02.021.
[7]孙天松,孟和毕力格,王俊国.新疆维吾尔自治区酸马奶化学组成与微生物学分析[J].中国乳品工业,2005,33(10):9-13.DOI:10.3969/j.issn.1001-2230.2005.10.002.
[8]布仁其其格.酸马奶传统发酵过程的宏转录组学分析及细菌群落结构动态变化研究[D].呼和浩特:内蒙古农业大学,2014.
[9]赵晓伟,丁君,窦妍,等.基于MiSeq测序技术分析红鳍东方鲀养殖环境菌群多样性[J].生态学杂志,2015,34(10):2965-2970.
[10]田颖刚,谢明勇,付志红,等.乌骨鸡脂肪油中脂肪酸组成的气相色谱-质谱分析[J].南昌大学学报(理科版),2006,30(3):264-266.
[11]陈洪兴.乳糖和乳糖不耐症[J].盐城工学院学报(自然科学版),2002,15(3):32-35.DOI:10.3969/j.issn.1671-5322.2002.03.010.
[12]JOST B,VILOTTE J L,DULUC I,et al.Production of lowlastose milk by ectopic expression of intestinal lactase in the mouse mammary gland[J].Nature Biotechnology,1999,17(2):160-164.DOI:10.1038/6158.
[13]额尔登孟克,潘琳,阿木尔图布兴,等.传统发酵乳制品酸马奶的研究进展[J].内蒙古农业大学学报(自然科学版),2016,37(2):134-139.
[14]拉·巴勒道尔吉,策·纳木斯来.蒙古艾日嘎[M].呼和浩特:内蒙古人民出版社,1990:137-138.
[15]布仁其其格,高雅罕,包艳青.酸马奶发酵过程中理化指标与细菌相的动态变化[J].食品工业科技,2016,37(11):118-121.
[16]ISHII S.Study on the kumiss(Airag)of Mongolian nomads after severe cold in the winters of 2000 and 2001[J].Milk Science,2003,52:49-52.
[17]郭本恒.功能性乳制品[M].北京:中国轻工业出版社,2001.
[18]ZHOU Y M,LIN J C,WU L P,et al.Effect of antecolic or retrocolic reconstruction of the gastro/duodenojejunostomy on delayed gastric emptying after pancreaticoduodenectomy:a meta-analysis[J].BMCGastroenterology,2015,15:68.DOI:10.1186/s12876-015-0300-8.
[19]QUIGLEY E M.Other forms of gastroparesis:postsurgical,parkinson,other neurologic diseases,connective tissue disorders[J].Gastroenterology Clinics of North America,2015,44(1):69-81.DOI:10.1016/j.gtc.2014.11.006.
[20]LACY B E.Functional dyspepsia and gastroparesis:one disease or two[J].American Journal of Gastroenterology,2012,107(11):1615-1620.DOI:10.1038/ajg.2012.104.
[21]PASRICHA P J.Future directions in the treatment of gastroparesis[J].Gastroenterology Clinics of North America,2015,44(1):185-189.DOI:10.1016/j.gtc.2014.11.014.
[22]SMITS M J,LOOTS C M,BENNINGA M A,et al.New insights in gastroesophageal reflux,esophageal function and gastric emptying in relation to dysphagia before and after anti-reflux surgery in children[J].Current Gastroenterology Reports,2013,15(10):351.DOI:10.1007/s11894-013-0351-3.
[23]GRASSI M,PETRACCIA L,MENNUNI G,et al.Changes,functional disorders,and diseases in the gastrointestinal tract of elderly[J].Nutricion Hospitalaria,2011,26(4):659-668.DOI:10.1590/S0212-16112011000400001.
[24]叶博.短链脂肪酸促进人和小鼠gastroids中产ghrelin细胞分化和小鼠胃排空的生理机制研究[D].上海:第二军医大学第一附属医院,2015.
[25]于昱,袁缨.多不饱和脂肪酸的营养研究[J].中国饲料,2003(24):21-23.DOI:10.3969/j.issn.1004-3314.2003.24.010.
[26]孙翔宇,高贵田,段爱莉,等.多不饱和脂肪酸的研究进展[J].食品工业科技,2012,33(7):418-423.DOI:10.13386/j.issn1002-0306.2012.07.030.
[27]孙天松,王俊国,张列兵,等.中国新疆地区酸马奶中乳酸菌生物多样性研究[J].微生物学通报,2007,34(3):451-454.DOI:10.3969/j.issn.0253-2654.2007.03.013.
[28]RING E,ANDERSEN R,SPERSTAD S,et al.Bacterial community of koumiss from Mongolia investigated by culture and cultureindependent methods[J].Food Biotechnology,2014,28(4):333-353.DOI:10.1080/08905436.2014.964253.
[29]Food and Agriculture Organization of United Nations,World Health Organization.Report on joint FAO/WHO expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria[S].Cordoba,Argentina,2001.
[30]李海燕,徐杰,云月英,等.传统发酵酸马奶中瑞士乳杆菌耐酸性的筛选[J].乳业科学与技术,2006,29(4):158-160.DOI:10.15922/j.cnki.jdst.2006.04.003.
[31]贾宏信,龚广予,郭本恒.益生菌干酪的研究进展[J].食品科学,2013,34(15):355-360.DOI:10.7506/spkx1002-6630-201315072.
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