混菌发酵苹果浊汁的益生菌筛选及其发酵动力学模型构建
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摘要
为筛选得到苹果浊汁发酵的较佳复合益生菌,以苹果浊汁为原料,从10种常见益生菌菌株中筛选出具有耐酸、耐胆盐特性及良好发酵性能的优良菌株,并进行复合菌株间拮抗实验以及复合菌株较佳比例的研究。此外,根据筛选得到的复合菌株较佳比例发酵苹果浊汁,构建菌体生长、产物生成及底物消耗动力学模型,研究确定模型的动力学参数,并进行验证。结果表明:发酵苹果浊汁的较佳复合菌株组合为嗜酸乳杆菌6005、植物乳杆菌21805、发酵乳杆菌21828,根据感官评定及活菌数得到3株菌株复合比例为1:1:1时,发酵苹果浊汁口感风味最佳,活菌数最高;建立复合菌株发酵苹果浊汁的菌体生长、产物生成和底物消耗动力学模型,模型理论值与实验值误差均小于10%,拟合较好,建立的动力学模型能够较好地预测复合益生菌发酵苹果浊汁发酵过程变化。
In order to select a suitable mixed culture of probiotics for cloudy apple juice fermentation, 10 strains of common probiotics were screened based on their acid and bile tolerance and fermentation performance. The antagonism between the selected strains was evaluated and the mixing ratio of these strains was determined. Furthermore, we modelled the kinetics of biomass growth, metabolite production and substrate consumption. The kinetic parameters of the models developed were determined and verified. The results showed that: 1) a mixture of Lactobacillus acidophilus 6005, Lactobacillus plantarum21805, and Lactobacillus fermentum 21828 was found to be the most suitable for the fermentation of apple juice at a ratio of 1:1:1 as determined based on sensory evaluation and viable cell counts, and the fermented apple juice had the best flavor and mouthfeel as well as the highest viable cell counts; and 2) the errors between the theoretical values from all these kinetic models and the experimental data were less than 10%, suggesting excellent goodness of fit. Hence, the kinetic models could predict the dynamic changes during the mixed culture fermentation of cloudy apple juice.
In order to select a suitable mixed culture of probiotics for cloudy apple juice fermentation, 10 strains of common probiotics were screened based on their acid and bile tolerance and fermentation performance. The antagonism between the selected strains was evaluated and the mixing ratio of these strains was determined. Furthermore, we modelled the kinetics of biomass growth, metabolite production and substrate consumption. The kinetic parameters of the models developed were determined and verified. The results showed that: 1) a mixture of Lactobacillus acidophilus 6005, Lactobacillus plantarum21805, and Lactobacillus fermentum 21828 was found to be the most suitable for the fermentation of apple juice at a ratio of 1:1:1 as determined based on sensory evaluation and viable cell counts, and the fermented apple juice had the best flavor and mouthfeel as well as the highest viable cell counts; and 2) the errors between the theoretical values from all these kinetic models and the experimental data were less than 10%, suggesting excellent goodness of fit. Hence, the kinetic models could predict the dynamic changes during the mixed culture fermentation of cloudy apple juice.
引文
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[21]许世闯,万嗣宝,李聪,等.乳酸菌发酵复合果蔬汁的菌种筛选及发酵工艺优化研究[J].食品科技,2017,42(9):2-7.DOI:10.13684/j.cnki.spkj.2017.09.001.
[22]KERBERG C,HOFVENDAHL K,ZACCHI G,et al.Modelling the influence of pH,temperature,glucose and lactic acid concentrations on the kinetics of lactic acid production by Lactococcus lactis ssp.lactis ATCC 19435 in whole-wheat flour[J].Applied Microbiology&Biotechnology,1998,49(6):682-690.DOI:10.1007/s002530051232.
[23]潘明,刘慧杰,郭脉玺,等.乳酸菌发酵荠菜-番茄复合蔬菜汁的动力学研究[J].食品科学,2009,30(23):281-285.DOI:10.3321/j.issn:1002-6630.2009.23.063.
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[25]ZACHAROF M P,LOVITT R W.Modelling and simulation of cell growth dynamics,substrate consumption,and lactic acid production kinetics of Lactococcus lactis[J].Biotechnology&Bioprocess Engineering,2013,18(1):52-64.DOI:10.1007/s12257-012-0477-4.
[26]李雪,白新鹏,曹君,等.仙人掌果酒发酵动力学及其抗氧化性[J].食品科学,2017,38(4):87-92.DOI:10.7506/spkx1002-6630-201704015.
[27]JIN H,LIU R,HE Y.Kinetics of batch fermentations for ethanol production with immobilized Saccharomyces cerevisiae growing on sweet sorghum stalk juice[J].Procedia Environmental Sciences,2012,12(8):137-145.DOI:10.1016/j.proenv.2012.01.258.
[28]DUARTE F N D,RODRIGUES J B,LIMA M D C,et al.Potential prebiotic properties of cashew apple(Anacardium occidentale L.)agro-industrial byproduct on Lactobacillus species[J].Journal of the Science of Food and Agriculture,2017,97(11):7-8.DOI:10.1002/jsfa.8232.
[29]BRAVO-FERRADA B M,HOLLMANN A,DELFEDERICO L,et al.Patagonian red wines:selection of Lactobacillus plantarum,isolates as potential starter cultures for malolactic fermentation[J].World Journal of Microbiology&Biotechnology,2013,29(9):1537-1549.DOI:10.1007/s11274-013-1337-x.
[30]CHARALAMPOPOULOS D,PANDIELLA S S,WEBB C.Growth studies of potentially probiotic lactic acid bacteria in cereal-based substrates[J].Journal of Applied Microbiology,2010,92(5):851-859.DOI:10.1046/j.1365-2672.2002.01592.x.
[31]NEMATOLLAHI A,SOHRABVANDI S,MORTAZAVIAN A M,et al.Viability of probiotic bacteria and some chemical and sensory characteristics in cornelian cherry juice during cold storage[J].Electronic Journal of Biotechnology,2016,21:49-53.DOI:10.1016/j.ejbt.2016.03.001.
[32]RICCI A,CIRLINI M,LEVANTE A,et al.Volatile profile of elderberry juice:Effect of lactic acid fermentation using L.plantarum,L.rhamnosus and L.casei strains[J].Food Research International,2018,105:412-422.DOI:10.1016/j.foodres.2017.11.042.
[33]李维妮,张宇翔,魏建平,等.益生菌发酵苹果汁工艺优化及有机酸的变化[J].食品科学,2017,38(22):80-87.DOI:10.7506/spkx1002-6630-201722013.
[2]熊涛.果蔬益生菌发酵关键技术与产业化应用[J].饮料工业,2016,19(5):71-73.DOI:10.3969/j.issn.1007-7871.2016.05.023.
[3]AFRC R F.Probiotics in man and animals[J].Journal of Applied Bacteriology,2010,66(5):365-378.DOI:10.1111/j.1365-2672.1989.tb05105.x.
[4]CORBO M R,BEVILACQUA A,PETRUZZI L,et al.Functional beverages:the emerging side of functional foods[J].Comprehensive Reviews in Food Science&Food Safety,2015,13(6):1192-1206.DOI:10.1111/1541-4337.12109.
[5]KANDYLIS P,PISSARIDI K,BEKATOROU A,et al.Dairy and non-dairy probiotic beverages[J].Current Opinion in Food Science,2016,7:58-63.DOI:10.1016/j.cofs.2015.11.012.
[6]KAPRASOB R,KERDCHOECHUEN O,LAOHAKUNJIT N,et al.Fermentation-based biotransformation of biaoctive phenolics and volatile compounds from cashew apple juice by select lactic acid bacteria[J].Process Biochemistry,2017,59:141-149.DOI:10.1016/j.procbio.2017.05.019.
[7]MOUSAVI Z E,MOUSAVI S M,RAZAVI S H,et al.Fermentation of pomegranate juice by probiotic lactic acid bacteria[J].World Journal of Microbiology&Biotechnology,2011,27(1):123-128.DOI:10.1007/s11274-010-0436-1.
[8]PANDA S K,BEHERA S K,QAKU X W,et al.Quality enhancement of prickly pears(Opuntia sp.)juice through probiotic fermentation using Lactobacillus fermentum-ATCC 9338[J].LWT-Food Science and Technology,2017,75:453-459.DOI:10.1016/j.lwt.2016.09.026.
[9]DIMITROVSKI D,VELICKOVA E,LANGERHOLC T,et al.Apple juice as a medium for fermentation by the probiotic Lactobacillus plantarum PCS 26 strain[J].Annals of Microbiology,2015,65(4):2161-2170.DOI:10.1007/s13213-015-1056-7.
[10]PEREIRA A L F,ALMEIDA F D L,JESUS A L T D,et al.Storage stability and acceptance of probiotic beverage from Cashew apple juice[J].Food&Bioprocess Technology,2013,6(11):3155-3165.DOI:10.1007/s11947-012-1032-1.
[11]李维妮,郭春锋,张宇翔,等.气相色谱-质谱法分析乳酸菌发酵苹果汁香气成分[J].食品科学,2017,38(4):146-154.DOI:10.7506/spkx1002-6630-201704024.
[12]颖宣,徐尔尼,冯乃宪,等.微生物混菌发酵应用研究进展[J].中国酿造,2008,27(9):1-4.DOI:10.3969/j.issn.0254-5071.2008.05.001.
[13]曹美霞.红肉苹果浊汁加工贮藏研究及工艺设计[D].西安:陕西师范大学,2016:1-2.
[14]赵光远,王璋,许时婴.混浊苹果汁加工过程中的酶促褐变及其防止的研究[J].食品工业科技,2003,24(10):57-61.DOI:10.3969/j.issn.1002-0306.2003.10.017.
[15]赵光远,王璋,许时婴.浑浊苹果汁加工中苹果PPO热失活条件的研究[J].食品科技,2003,28(7):84-87;95.DOI:10.13684/j.cnki.spkj.2003.07.027.
[16]国家质量监督检验检疫总局.食品中总酸的测定:GB/T 12456-2008[S].北京:中国标准出版社,2008:1-2.
[17]国内贸易部食品检测科学研究所.果汁通用试验方法:SB/T10203-1994[S].北京:中国标准出版社,1994:48-49.
[18]国家食品药品监督管理总局.食品微生物学检验乳酸菌检验:GB 4789.35-2016[S].北京:中国标准出版社,2016:1-8.
[19]VERóN H E,RISIO H D D,ISLA M I,et al.Isolation and selection of potential probiotic lactic acid bacteria from Opuntia ficus-indica,fruits that grow in Northwest Argentina[J].LWT-Food Science and Technology,2017,84:3-4.DOI:10.1016/j.lwt.2017.05.058.
[20]宋苏华.发酵果蔬饮料益生乳酸菌的强化筛选及其发酵工艺技术研究[D].南昌:南昌大学,2012:21-22.
[21]许世闯,万嗣宝,李聪,等.乳酸菌发酵复合果蔬汁的菌种筛选及发酵工艺优化研究[J].食品科技,2017,42(9):2-7.DOI:10.13684/j.cnki.spkj.2017.09.001.
[22]KERBERG C,HOFVENDAHL K,ZACCHI G,et al.Modelling the influence of pH,temperature,glucose and lactic acid concentrations on the kinetics of lactic acid production by Lactococcus lactis ssp.lactis ATCC 19435 in whole-wheat flour[J].Applied Microbiology&Biotechnology,1998,49(6):682-690.DOI:10.1007/s002530051232.
[23]潘明,刘慧杰,郭脉玺,等.乳酸菌发酵荠菜-番茄复合蔬菜汁的动力学研究[J].食品科学,2009,30(23):281-285.DOI:10.3321/j.issn:1002-6630.2009.23.063.
[24]后立琼.苗族酸汤中乳酸菌的分离鉴定及发酵动力学模型研究[D].成都:四川农业大学,2012:33-34.
[25]ZACHAROF M P,LOVITT R W.Modelling and simulation of cell growth dynamics,substrate consumption,and lactic acid production kinetics of Lactococcus lactis[J].Biotechnology&Bioprocess Engineering,2013,18(1):52-64.DOI:10.1007/s12257-012-0477-4.
[26]李雪,白新鹏,曹君,等.仙人掌果酒发酵动力学及其抗氧化性[J].食品科学,2017,38(4):87-92.DOI:10.7506/spkx1002-6630-201704015.
[27]JIN H,LIU R,HE Y.Kinetics of batch fermentations for ethanol production with immobilized Saccharomyces cerevisiae growing on sweet sorghum stalk juice[J].Procedia Environmental Sciences,2012,12(8):137-145.DOI:10.1016/j.proenv.2012.01.258.
[28]DUARTE F N D,RODRIGUES J B,LIMA M D C,et al.Potential prebiotic properties of cashew apple(Anacardium occidentale L.)agro-industrial byproduct on Lactobacillus species[J].Journal of the Science of Food and Agriculture,2017,97(11):7-8.DOI:10.1002/jsfa.8232.
[29]BRAVO-FERRADA B M,HOLLMANN A,DELFEDERICO L,et al.Patagonian red wines:selection of Lactobacillus plantarum,isolates as potential starter cultures for malolactic fermentation[J].World Journal of Microbiology&Biotechnology,2013,29(9):1537-1549.DOI:10.1007/s11274-013-1337-x.
[30]CHARALAMPOPOULOS D,PANDIELLA S S,WEBB C.Growth studies of potentially probiotic lactic acid bacteria in cereal-based substrates[J].Journal of Applied Microbiology,2010,92(5):851-859.DOI:10.1046/j.1365-2672.2002.01592.x.
[31]NEMATOLLAHI A,SOHRABVANDI S,MORTAZAVIAN A M,et al.Viability of probiotic bacteria and some chemical and sensory characteristics in cornelian cherry juice during cold storage[J].Electronic Journal of Biotechnology,2016,21:49-53.DOI:10.1016/j.ejbt.2016.03.001.
[32]RICCI A,CIRLINI M,LEVANTE A,et al.Volatile profile of elderberry juice:Effect of lactic acid fermentation using L.plantarum,L.rhamnosus and L.casei strains[J].Food Research International,2018,105:412-422.DOI:10.1016/j.foodres.2017.11.042.
[33]李维妮,张宇翔,魏建平,等.益生菌发酵苹果汁工艺优化及有机酸的变化[J].食品科学,2017,38(22):80-87.DOI:10.7506/spkx1002-6630-201722013.