传统发酵食品中乳酸菌的分离鉴定、基因分型及优良菌株筛选
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摘要
对分离自传统发酵食品的40株乳酸菌进行16S r DNA鉴定、随机扩增多态性DNA(RAPD)菌株基因分型及产酸、产乙偶姻、降胆固醇、分解亚硝酸盐及1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除能力进行研究。结果表明,40株乳酸菌中,20株植物乳杆菌(Lactobacillus plantarum),分为Ⅰ、Ⅱ、Ⅲ、Ⅳ4种型,Ⅲ型又分2个亚型;8株短乳杆菌(Lactobacillus brevis)、6株戊糖片球菌(Pediococcus pentosaceus)均呈现Ⅰ、Ⅱ、Ⅲ3种型,而6株耐久肠球菌(Enterococcus durans)呈现Ⅰ、Ⅱ2种型,显示了传统发酵食品中乳酸菌菌株的基因多样性。同种同型乳酸菌产酸、乙偶姻及DPPH自由基清除能力具有相似性;但同种不同型具有差异性。最终筛选出1株产酸(24.3 g/L)、乙偶姻(0.54 mg/m L)、分解亚硝酸盐(74.15%)、DPPH自由基清除能力(92.29%)均较强的植物乳杆菌SAV-JL7,以期为发酵食品风味及品质的改善提供参考。
40 strains of lactic acid bacteria isolated from traditional fermented food were identified by 16 S r DNA, and genotyping by random amplified polymorphic DNA(RAPD). The acid production, acetoin production, cholesterol lowering, nitrite decomposition and DPPH free radical scavenging capacity were studied. The results showed that among the 40 lactic acid bacteria strains, 20 strains were Lactobacillus plantarum, and they were divided into I, II, III and IV types, and type III was divided into 2 subtypes; in addition, 8 strains were Lactobacillus brevis, 6 strains were Pediococcus pentosaceus, and they were all divided into 3 types I, II, III; 6 strains were Enterococcus durans, and they were all divided into 2 types I, II, which showed that the lactic acid bacteria isolated from traditional fermented food had genetic diversity. The same type of lactic acid bacteria had the similarity in acid production, acetoin production and DPPH free radical scavenging ability; different types were different in characteristic. A strain of L. plantarum SAV-JL7 with strong acid production(24.3 g/L), acetoin production(0.54 mg/ml), nitrite decomposition(74.15%) and DPPH free radical scavenging ability(92.29%) was screened finally, in order to provide reference for the improvement of the flavor and quality of the fermented food.
40 strains of lactic acid bacteria isolated from traditional fermented food were identified by 16 S r DNA, and genotyping by random amplified polymorphic DNA(RAPD). The acid production, acetoin production, cholesterol lowering, nitrite decomposition and DPPH free radical scavenging capacity were studied. The results showed that among the 40 lactic acid bacteria strains, 20 strains were Lactobacillus plantarum, and they were divided into I, II, III and IV types, and type III was divided into 2 subtypes; in addition, 8 strains were Lactobacillus brevis, 6 strains were Pediococcus pentosaceus, and they were all divided into 3 types I, II, III; 6 strains were Enterococcus durans, and they were all divided into 2 types I, II, which showed that the lactic acid bacteria isolated from traditional fermented food had genetic diversity. The same type of lactic acid bacteria had the similarity in acid production, acetoin production and DPPH free radical scavenging ability; different types were different in characteristic. A strain of L. plantarum SAV-JL7 with strong acid production(24.3 g/L), acetoin production(0.54 mg/ml), nitrite decomposition(74.15%) and DPPH free radical scavenging ability(92.29%) was screened finally, in order to provide reference for the improvement of the flavor and quality of the fermented food.
引文
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[21]周海柱,高云航,郭玮,等.高效降胆固醇植物乳杆菌的筛选及其益生潜能初探[J].中国农业大学学报,2018,23(2):36-42.
[22]FEI Y T,LIU D M,LUO L H,et al.Molecular characterization of Lactobacillus plantarum DMDL 9010,a strain with efficient nitrite degradation capacity[J].Plos One,2014,9(11):113792.
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[24]程方方,武运,武亚婷,等.酸马乳源降解亚硝酸盐乳酸菌的分离及鉴定[J].中国酿造,2018,37(9):72-77.
[25]韩墨,王燕,杨志鹏,等.内蒙古传统酸奶乳酸菌的筛选及体外益生效果评价[J].食品研究与开发,2018(1):152-156.
[2]POGAI T,MANCINI A,SANTARELLI M,et al.Diversity and dynamic of lactic acid bacteria strains during aging of a long ripened hard cheese produced from raw milk and undefined natural starter[J].Food Microbiol,2013,36(2):207-215.
[3]ZARE E M,LASHANI E,DAVOODABADI A.Antimicrobial properties of lactic acid bacteria isolated from traditional yogurt and milk against Shigella strains[J].Gms Hygiene Infect Control,2018,13(1):1-5.
[4]TAKUYA Y,TATSUJI S,TAKENORI N,et al.Lactic acid bacteria from kefir increase cytotoxicity of natural killer cells to tumor cells[J].Foods,2018,7(4):48-56.
[5]LIU A,LI X,PU B,et al.Use of psychrotolerant lactic acid bacteria isolated from Chinese traditional Paocai for the quality improvement of Paocai products[J].J Agr Food Chem,2017,65(12):2580-2587.
[6]焦玉双.山西老陈醋大曲中优良乳酸菌的分离筛选及其发酵应用研究[D].晋中:山西农业大学,2016.
[7]LU Z M,LIU N,WANG L J,et al.Elucidating and regulating the acetoin producing role of microbial functional group in multispecies acetic acid fermentation[J].Appl Environ Microbiol,2016,82(19):5860-5868.
[8]KUNG L,SMITH M L,ERICA B D S,et al.An evaluation of the effectiveness of a chemical additive based on sodium benzoate,potassium sorbate,and sodium nitrite on the fermentation and aerobic stability of corn silage[J].J Dairy Sci,2018,101(7):5949-5960.
[9]黄燕燕,郭均,黎恒希,等.降胆固醇乳酸菌的体外筛选及其降胆固醇机理探讨[J].食品科学,2018,39(6):88-94.
[10]WANG L C,PAN T M,TSAI T Y.Lactic acid bacteria-fermented product of green tea and Houttuynia cordata leaves exerts anti-adipogenic and anti-obesity effects[J].J Food Drug Anal,2018,26(3):973.
[11]WULLCHLEGER S,LACROIX C,BONFOH B,et al.Analysis of lactic acid bacteria communities and their seasonal;variations in a spontaneously fermented dairy product(Malian fene)by applying a cultivation/genotype-based binary model[J].Int Dairy J,2013,29(1):28-35.
[12]RANDAZZO C L,RESTUCCIA C,ROMANO A D,et al.Lactobacillus casei,dominant species in naturally fermented Sicilian green olives[J].Int J Food Microbiol,2004,90(1):9-14.
[13]PSONI L,KOTZAMANIDIS C,YIANGOU M,et al.Genotypic and phenotypic diversity of Lactococcus lactis isolates from Batzos,a Greek PDO raw goat milk cheese[J].Int J Food Microbiol,2007,114(2):211-220.
[14]DIMITROV Z P,MINKOVA S,MICHAYLOVA M.Comparative evaluation of three molecular typing methods in their applicability to differentiate Lactobacillus strains with human origin[J].World J Microbiol Biot,2008,24(8):1305-1312.
[15]ADIGUZEL G C,ATASEVER M.Phenotypic and genotypic characterization of lactic acid bacteria isolated from Turkish dry fermented sausage[J].Roman Biotechnol Lett,2009,14(1):4130-4138.
[16]周宁.中国不同地区市售酸奶中乳酸菌的药物敏感性分析[D].杨凌:西北农林科技大学,2012.
[17]李艳莉.酸奶及发酵剂中分离乳酸菌药物敏感性研究[D].广州:华南理工大学,2015.
[18]吴林寰.整合型元基因组序列分析平台构建及其在酿醋微生物群落功能研究中的应用[D].无锡:江南大学,2016.
[19]WANG S C,CHANG C K,CHAN S C,et al.Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol[J].Asian Pac J Trop Biomed,2014,4(7):523-528.
[20]黄颖,隋玉杰,钟莉莉,等.藏灵菇发酵乳中降胆固醇乳酸菌的分离及鉴定[J].吉林大学学报,2010,36(6):1181-1185.
[21]周海柱,高云航,郭玮,等.高效降胆固醇植物乳杆菌的筛选及其益生潜能初探[J].中国农业大学学报,2018,23(2):36-42.
[22]FEI Y T,LIU D M,LUO L H,et al.Molecular characterization of Lactobacillus plantarum DMDL 9010,a strain with efficient nitrite degradation capacity[J].Plos One,2014,9(11):113792.
[23]荣凤君.四种自制发酵食品中益生乳杆菌的筛选及功能特性研究[D].长春:吉林农业大学,2017.
[24]程方方,武运,武亚婷,等.酸马乳源降解亚硝酸盐乳酸菌的分离及鉴定[J].中国酿造,2018,37(9):72-77.
[25]韩墨,王燕,杨志鹏,等.内蒙古传统酸奶乳酸菌的筛选及体外益生效果评价[J].食品研究与开发,2018(1):152-156.