高效降解嘌呤核苷乳酸菌的筛选及特性研究
|
摘要
采用乳酸菌对高尿酸血症及其相关病症进行辅助治疗,可以减轻严格饮食限制(传统治疗方法)对患者生活质量的影响。本研究以市售泡菜为材料,用HPLC法测定乳酸菌对肌苷和鸟苷的降解率,筛选出具有高效降解嘌呤核苷能力的乳酸菌。在筛选菌株中,菌株5-1的降解能力最强,对肌苷和鸟苷的降解率分别为89.37%和93.20%。该菌株经生理生化和16S rDNA测序鉴定为弯曲乳杆菌(Lactobacillus curvatus)。在pH 3或胆盐浓度0.3%的条件下处理4 h后,Lb.curvatus 5-1活菌数保持在107cfu/mL以上。另外,该菌株在人工胃肠液中展现出良好的耐受性,在肠液中的核苷降解能力与在缓冲液中的相当。因此,Lb. curvatus 5-1是开发高尿酸血症预防及辅助治疗的微生态菌剂的优良菌株,具有良好的应用前景。
Lactic acid bacteria(LAB) can be used for prevention and auxiliary treatment of hyperuricemia and its complications. By using such way,the detrimental effect of people's living quality caused by maintaining strict dietary restriction(the conventional treatment of hyperuricemia)could be reduced or eliminated. In this study the purine nucleoside-decomposing ability of LABs were isolated from Chinese sauerkraut bought from local supermarket by HPLC method through the determination of their insosine and guanosine degradation abilities. Among which strain 5-1 showed the best probiotic potential with degradation rate of insosine and guanosine are 89.37% and 93.20%respectively. Based on physiological,biochemical tests and 16 S rDNA classification,strain 5-1 was identified as Lactobacillus curvatus. After 4 h treatment under the circumstance of pH 3 or 0.3% bile salt,strain 5-1 could keep a survival level of 107 cfu/mL. Furthermore the great tolerance capacity was shown in the artificial gastrointestinal fluid with food suspension and excellent purine nucleoside-decomposing ability in artificial intestinal juice with food suspension.Therefore,Lb.curvatus 5-1 has a great potential for applications in the development of microbial agents that help to prevent and auxiliarily treat hyperuricemia.
Lactic acid bacteria(LAB) can be used for prevention and auxiliary treatment of hyperuricemia and its complications. By using such way,the detrimental effect of people's living quality caused by maintaining strict dietary restriction(the conventional treatment of hyperuricemia)could be reduced or eliminated. In this study the purine nucleoside-decomposing ability of LABs were isolated from Chinese sauerkraut bought from local supermarket by HPLC method through the determination of their insosine and guanosine degradation abilities. Among which strain 5-1 showed the best probiotic potential with degradation rate of insosine and guanosine are 89.37% and 93.20%respectively. Based on physiological,biochemical tests and 16 S rDNA classification,strain 5-1 was identified as Lactobacillus curvatus. After 4 h treatment under the circumstance of pH 3 or 0.3% bile salt,strain 5-1 could keep a survival level of 107 cfu/mL. Furthermore the great tolerance capacity was shown in the artificial gastrointestinal fluid with food suspension and excellent purine nucleoside-decomposing ability in artificial intestinal juice with food suspension.Therefore,Lb.curvatus 5-1 has a great potential for applications in the development of microbial agents that help to prevent and auxiliarily treat hyperuricemia.
引文
[1]GLIOZZI M,MALARA N,MUSCOLI S,et al.The treatment of hyperuricemia[J].International Journal of Cardiology,2016,213:23-27.
[2]YU S S,YANG H M,GUO X F,et al.Prevalence of hyperuricemia and its correlates in rural Northeast Chinese population:from lifestyle risk factors to metabolic comorbidities[J].Clinical Rheumatology,2016,35(5):1207-1215.
[3]ZENG Xuejun.Interpretation of the Chinese guideline for the management of gout in 2010[J].Chinese Journal of Practical Internal Medicine,2012,32(6):438-441.(in Chinese)
[4]ZHANG Chen,LI Junling,TIAN Jiaxing,et al.Research progress of treating hyperuricemia by traditional Chinese medicine[J].Chinese Journal of New Drugs,2013,22(6):670-673.(in Chinese)
[5]YAN D D,WANG J,JIANG F,et al.A causal relationship between uric acid and diabetic macrovascular disease in Chinese type2 diabetes patients:A Mendelian randomization analysis[J].International Journal of Cardiology,2016,214:194-199.
[6]HARRIS M D,SIEGEL L B,ALLOWAY J A.Gout and hyperuricemia[J].American Family Physician,1999,59(4):925-934.
[7]VILLEGAS R,XIANG Y B,ELASY T,et al.Purine-rich foods,protein intake,and the prevalence of hyperuricemia:The Shanghai Men’s Health Study[J].Nutrition Metabolism and Cardiovascular Diseases,2012,22(5):409-416.
[8]YAMADA N,IWAMOTO C,NAKAMURA M,et al.Reducing effect of Lactobacillus gasseri PA-3 on the absorption of foodderived purines[J].Milk Science,2016,65(1):25-31.
[9]TSUBOI H,KANEKO N,SATOU A,et al.Lactic acid bacteria having action of lowering blood uric acid level:美国,USO8460918[P].2013-06-11.
[10]TSUBOI H,KANEKO N,SATOU A,et al.Lactic acid bacterium having effect of lowering blood uric acid level:欧洲,EP2221360A1[P].201-08-25.
[11]YANG Mingjun,FENG Huiqin,YANG Qingyao,et al.HPLC determination of nucleosides of Hirsutella sinensis mycelium[J].Food Research and Development,2011,32(11):96-99.(in Chinese)
[12]BAO Qiuhua,LV Qiang,YU Jie,et al.Molecular identification of nine strains lactic acid bacteria isolatedfrom yak milk and triton in Sichuan[J].Journal of Food Science and Biotechnology,2012,31(4):396-401.(in Chinese)
[13]KHALIL M A,EI-SHEEKH M M,EI-ADAWL H I,et al.Efficacy of microencapsulated lactic acid bacteria in Helicobacter pylori eradication therapy[J].Journal of Research in Medical Sciences,2015,20(10):950-957.
[14]FENG Dawei,ZHOU Jiachun.Evaluation of agar disc diffusion method for determining antibiotic susceptibility of lactic acid bacteria strains by a selected culture medium[J].Microbiology China,2010,37(3):454-464.(in Chinese)
[15]CHANG L,ZHUO Y Z,KE D,et al.Antibiotic resistance of probiotic strains of lactic acid bacteria isolated from marketed foods and drugs[J].Biomedical and Environmental Sciences,2009,22(5):401-412.
[16]ERKKILA S,PETAJA E.Screening of commercial meat starter cultures at low pH and in the presence of bile salts for potential probiotic use[J].Meat Science,2000,55(3):297-300.
[17]CHOU L S,WEIMER B.Isolation and characterization of acid-and bile-tolerant isolates from strains of Lactobacillus acidophilus[J].Journal of Dairy Science,1999,82(1):23-31.
[18]郭兴华,凌代文,王安如,等.乳酸细菌现代研究实验技术[M].北京:科学出版社,2013:312-314.
[19]SANDERS M E,WALKER D C,WALKER K M,et al.Performance of commercial cultures in fluid milk applications[J].Journal of dairy science,1996,79(6):943-955.
[20]GILLILAND S E.Health and nutritional benefits from lactic acid bacteria[J].FEMS Microbiology Letters,1990,87(1):175-188.
[2]YU S S,YANG H M,GUO X F,et al.Prevalence of hyperuricemia and its correlates in rural Northeast Chinese population:from lifestyle risk factors to metabolic comorbidities[J].Clinical Rheumatology,2016,35(5):1207-1215.
[3]ZENG Xuejun.Interpretation of the Chinese guideline for the management of gout in 2010[J].Chinese Journal of Practical Internal Medicine,2012,32(6):438-441.(in Chinese)
[4]ZHANG Chen,LI Junling,TIAN Jiaxing,et al.Research progress of treating hyperuricemia by traditional Chinese medicine[J].Chinese Journal of New Drugs,2013,22(6):670-673.(in Chinese)
[5]YAN D D,WANG J,JIANG F,et al.A causal relationship between uric acid and diabetic macrovascular disease in Chinese type2 diabetes patients:A Mendelian randomization analysis[J].International Journal of Cardiology,2016,214:194-199.
[6]HARRIS M D,SIEGEL L B,ALLOWAY J A.Gout and hyperuricemia[J].American Family Physician,1999,59(4):925-934.
[7]VILLEGAS R,XIANG Y B,ELASY T,et al.Purine-rich foods,protein intake,and the prevalence of hyperuricemia:The Shanghai Men’s Health Study[J].Nutrition Metabolism and Cardiovascular Diseases,2012,22(5):409-416.
[8]YAMADA N,IWAMOTO C,NAKAMURA M,et al.Reducing effect of Lactobacillus gasseri PA-3 on the absorption of foodderived purines[J].Milk Science,2016,65(1):25-31.
[9]TSUBOI H,KANEKO N,SATOU A,et al.Lactic acid bacteria having action of lowering blood uric acid level:美国,USO8460918[P].2013-06-11.
[10]TSUBOI H,KANEKO N,SATOU A,et al.Lactic acid bacterium having effect of lowering blood uric acid level:欧洲,EP2221360A1[P].201-08-25.
[11]YANG Mingjun,FENG Huiqin,YANG Qingyao,et al.HPLC determination of nucleosides of Hirsutella sinensis mycelium[J].Food Research and Development,2011,32(11):96-99.(in Chinese)
[12]BAO Qiuhua,LV Qiang,YU Jie,et al.Molecular identification of nine strains lactic acid bacteria isolatedfrom yak milk and triton in Sichuan[J].Journal of Food Science and Biotechnology,2012,31(4):396-401.(in Chinese)
[13]KHALIL M A,EI-SHEEKH M M,EI-ADAWL H I,et al.Efficacy of microencapsulated lactic acid bacteria in Helicobacter pylori eradication therapy[J].Journal of Research in Medical Sciences,2015,20(10):950-957.
[14]FENG Dawei,ZHOU Jiachun.Evaluation of agar disc diffusion method for determining antibiotic susceptibility of lactic acid bacteria strains by a selected culture medium[J].Microbiology China,2010,37(3):454-464.(in Chinese)
[15]CHANG L,ZHUO Y Z,KE D,et al.Antibiotic resistance of probiotic strains of lactic acid bacteria isolated from marketed foods and drugs[J].Biomedical and Environmental Sciences,2009,22(5):401-412.
[16]ERKKILA S,PETAJA E.Screening of commercial meat starter cultures at low pH and in the presence of bile salts for potential probiotic use[J].Meat Science,2000,55(3):297-300.
[17]CHOU L S,WEIMER B.Isolation and characterization of acid-and bile-tolerant isolates from strains of Lactobacillus acidophilus[J].Journal of Dairy Science,1999,82(1):23-31.
[18]郭兴华,凌代文,王安如,等.乳酸细菌现代研究实验技术[M].北京:科学出版社,2013:312-314.
[19]SANDERS M E,WALKER D C,WALKER K M,et al.Performance of commercial cultures in fluid milk applications[J].Journal of dairy science,1996,79(6):943-955.
[20]GILLILAND S E.Health and nutritional benefits from lactic acid bacteria[J].FEMS Microbiology Letters,1990,87(1):175-188.