不同品种豇豆发酵过程中质构品质变化及产植物细胞壁降解酶微生物种类分析
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摘要
以4种不同品种豇豆为研究对象,探究发酵过程中菌群和质构特性变化,分析造成不同品种发酵豇豆软腐的产植物细胞壁降解酶(Plant cell wall degradation enzymes,PCWDEs)微生物的主要种类。采用质构分析仪测定发酵过程中豇豆质构的变化,用鉴别培养基筛选产PCWDEs微生物并进行16S rDNA或18S rDNA分子生物学鉴定。结果表明:豇豆品种A(成豇10号)在发酵过程中表现出的质构品质最好,与豇豆品种A相比,豆品种B(红嘴胖豇)、品种C(成豇9号)和品种D(挂面红)的硬度、弹性、咀嚼力下降得更快,尤其是豇豆品种B。对产PCWDES微生物种类分析得知,品种B发酵豇豆中产PCWDEs微生物数量和种类最多且产酶类别多样,微生物分属于肠杆菌属(Enterobacter)、柠檬酸杆菌属(Citrobacter)、假单胞菌属(Pseudomonas)、葡萄球菌属(Staphylococcus)、粘菌属(Myroides)、泛菌属(Pantoea)和克雷伯氏菌属(Klebsiella)等属;品种A、品种C、品种D中产PCWDEs微生物及产酶种类较少,仅在品种D中发现产PCWDEs的解鸟氨酸拉乌尔菌(Raoultella ornithinolytica); 4种发酵豇豆中均分离得到产蛋白酶的植物乳杆菌(Lactobacillus plantarum)和产淀粉酶的异常威克汉姆酵母(Wickerhamomyces anomalus)。综上所述,不同品种豇豆在发酵过程中微生物群体及质构品质变化具有品种特异性,发酵后期豇豆的软腐与产PCWDEs微生物的种类和数量有关。
Four different varieties of cowpea were studied as the research object,and the changes of microbial flora and texture properties were explored.The main typical species of plant cell wall degrading enzymes( PCWDEs) caused by different varieties of fermented cowpea soft rot were analyzed.The textures of fermented cowpea were determined by texture analyzer.The PCWDEs producing agents were separated by differential plates and identified through the 16S/18S rDNA sequencing.The results showed that cowpea A presented the best texture quality during the whole fermenting process.The hardness,springness and chewiness of cowpea B,cowpea C,cowpea D were destroyed faster than cowpea A in fermentation process,especially cowpea B. Cell wall degrading enzymes( PCWDEs) producing isolates were selected.The results presented that the fermented cowpea B harbored the most numbers and varieties of PCWDEs producing microorganisms,which belonged to Enterobacter,Citrobacter,Pseudomonas,Staphylococcus,Myroides,Pantoea and Klebsiella.The PCWDEs microorganisms in fermented cowpea A,cowpea C and cowpea D were less than cowpea B. Only Raoultella ornithinolytica presented in cowpea D. Lactobacillus plantarum producing protease and Wickerhamomyces anomalus producing amylase were found in four fermented cowpeas. The present study showed that the texture and microbial changes during the fermentation in different fermented cowpeas presented respective specificities.The softrot of fermented cowpeas was related to the number and variety of PCWDEs producing agents in the later fermented process.
Four different varieties of cowpea were studied as the research object,and the changes of microbial flora and texture properties were explored.The main typical species of plant cell wall degrading enzymes( PCWDEs) caused by different varieties of fermented cowpea soft rot were analyzed.The textures of fermented cowpea were determined by texture analyzer.The PCWDEs producing agents were separated by differential plates and identified through the 16S/18S rDNA sequencing.The results showed that cowpea A presented the best texture quality during the whole fermenting process.The hardness,springness and chewiness of cowpea B,cowpea C,cowpea D were destroyed faster than cowpea A in fermentation process,especially cowpea B. Cell wall degrading enzymes( PCWDEs) producing isolates were selected.The results presented that the fermented cowpea B harbored the most numbers and varieties of PCWDEs producing microorganisms,which belonged to Enterobacter,Citrobacter,Pseudomonas,Staphylococcus,Myroides,Pantoea and Klebsiella.The PCWDEs microorganisms in fermented cowpea A,cowpea C and cowpea D were less than cowpea B. Only Raoultella ornithinolytica presented in cowpea D. Lactobacillus plantarum producing protease and Wickerhamomyces anomalus producing amylase were found in four fermented cowpeas. The present study showed that the texture and microbial changes during the fermentation in different fermented cowpeas presented respective specificities.The softrot of fermented cowpeas was related to the number and variety of PCWDEs producing agents in the later fermented process.
引文
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[17]陆宽,王雪雅,霍昕,等.电子鼻结合顶空固相微萃取-气质联用技术分析贵州不同品种辣椒发酵后挥发性成分[J].食品科学,2018,39(4):199-205.
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[19]陈玲,冯宽,向娟,等.不同品种及货架期对泡豇豆质量的影响[J].食品科技,2017,42(10):48-51.
[20]周情操.豇豆泡制加工适性评价及脆性研究[D].武汉:华中农业大学,2007.
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[22]Wu R,Yu M,Liu X,et al.Changes in flavour and microbial diversity during natural fermentation of suan-cai,a traditional food made in Northeast China[J].International Journal of Food Microbiology,2015,211:23-31.
[23]刘春燕,戴明福,夏姣,等.不同乳酸菌接种发酵泡菜风味的研究[J].食品工业科技,2015,36(7):154-158.
[24]Tamang J P,Tamang B,Schillinger U,et al.Identification of predominant lactic acid bacteria isolated from traditionally fermented vegetable products of the Eastern Himalayas[J].International Journal of Food Microbiology,2005,105(3):347-356.
[25]Johanningsmeier S D,Mcfeeters R F.Metabolism of lactic acid in fermented cucumbers by Lactobacillus buchneri and related species,potential spoilage organisms in reduced salt fermentations[J].Food Microbiology,2013,35(2):129-135.
[26]Franco W,Pérez-díaz I M.Role of selected oxidative yeasts and bacteria in cucumber secondary fermentation associated with spoilage of the fermented fruit[J].Food Microbiology,2012,32(2):338-344.
[27]Laitila A,Sarlin T,Raulio M,et al.Yeasts in malting,with special emphasis on Wickerhamomyces anomalus(synonym Pichia anomala)[J].Antonie Van Leeuwenhoek,2011,99(1):75-84.
[28]张腾.发酵蔬菜微生物群落变化及与亚硝酸盐的关系[D].太原:山西大学,2013.
[29]Krishnan P,Bhat R,Kush A,et al.Isolation and functiona characterization of bacterial endophytes from Carica papaya fruits[J].Journal of Applied Microbiology,2012,113(2):308-317.
[30]Rao Y,Qian Y,She X,et al.Pellicle formation,microbia succession and lactic acid utilisation during the aerobic deteriorating process of Sichuan pickle[J].International Journal of Food Science and Technology,2017,53(3).
[31]Zogaj X,Bokranz W,Nimtz M,et al.Production of cellulose and curli fimbriae by members of the family Enterobacteriaceae isolated from the human gastrointestinal tract[J].Infection and Immunity,2003,71(7):4151-4158.
[2]梁莉,杜阿如娜,马涛,等.低盐豇豆泡菜预处理工艺优化及贮藏特性分析[J].食品科学,2018,39(6):246-251.
[3]Kapravelou G,Martínez R,Andrade A M,et al.Improvemen of the antioxidant and hypolipidaemic effects of cowpea flours,by fermentation:Results of and experiments[J].Journal of the Science of Food&Agriculture,2015,95(6):1207-1216.
[4]Caplice E,Fitzgerald G F.Food fermentations:Role o microorganisms in food production and preservation[J].International Journal of Food Microbiology,1999,50(1-2):131-149.
[5]何鹏晖,钱杨,王猛,等.腐败发酵蔬菜中产膜醭细菌的分离鉴定及其生长特性分析[J].食品科学,2017,38(10):92-97.
[6]董章勇,王振中.植物病原真菌细胞壁降解酶的研究进展[J].湖北农业科学,2012,51(21):4697-4700.
[7]谢占玲.植物细胞壁降解真菌及酶的研究[D].兰州:甘肃农业大学,2011.
[8]周情操,徐跃进,潘思轶.不同腌制条件下豇豆品质变化规律研究[J].食品科学,2006,27(10):122-125.
[9]Lee D H,Kim J B,Kim M,et al.Microbiota on spoiled vegetables and their characterization[J].Journal of Food Protection,2013,76(8):1350-1358.
[10]蒋云露,杨建涛,王猛,等.四川泡菜盐卤中微生物总基因组DNA提取方法的比较[J].中国酿造,2015,34(4):90-92.
[11]饶瑜,常伟,龚丽,等.四川泡菜生花酵母的分离与鉴定[J].食品与发酵科技,2013,49(3):19-22.
[12]Franco W,Perez-diaz I M.Microbial interactions associated with secondary cucumber fermentation[J].Journal of Applied Microbiology,2013,114(1):161-172.
[13]Rao Y,Wang M,Jiang Y L,et al.Isolation of bacteriocinogenic lactic acid bacteria for biopreservation of Chinese traditional Sichuan pickle[J].食品科学,2015,36(3):171-177.
[14]何鹏晖,厍晓,钱杨,等.发酵蔬菜中腐败微生物及其防控的研究进展[J].食品工业科技,2017,38(11):374-384.
[15]熊涛,彭飞,李啸,等.传统发酵泡菜优势微生物及其代谢特性[J].食品科学,2015,36(3):158-161.
[16]黎庭耀,李桂花,陈汉才,等.不同豇豆品种资源的营养品质分析[J].广东农业科学,2017,44(4):32-37.
[17]陆宽,王雪雅,霍昕,等.电子鼻结合顶空固相微萃取-气质联用技术分析贵州不同品种辣椒发酵后挥发性成分[J].食品科学,2018,39(4):199-205.
[18]张雁,黄丽慧,陈于陇,等.不同品种芥菜发酵过程中亚硝酸盐变化规律的研究[J].现代食品科技,2013,29(9):2152-2157.
[19]陈玲,冯宽,向娟,等.不同品种及货架期对泡豇豆质量的影响[J].食品科技,2017,42(10):48-51.
[20]周情操.豇豆泡制加工适性评价及脆性研究[D].武汉:华中农业大学,2007.
[21]Zhao N,Zhang C,Yang Q,et al.Multiple roles of lactic acid bacteria microflora in the formation of marker flavour compounds in traditional chinese paocai[J].Rsc Advances,2016,6(92):89671-89678.
[22]Wu R,Yu M,Liu X,et al.Changes in flavour and microbial diversity during natural fermentation of suan-cai,a traditional food made in Northeast China[J].International Journal of Food Microbiology,2015,211:23-31.
[23]刘春燕,戴明福,夏姣,等.不同乳酸菌接种发酵泡菜风味的研究[J].食品工业科技,2015,36(7):154-158.
[24]Tamang J P,Tamang B,Schillinger U,et al.Identification of predominant lactic acid bacteria isolated from traditionally fermented vegetable products of the Eastern Himalayas[J].International Journal of Food Microbiology,2005,105(3):347-356.
[25]Johanningsmeier S D,Mcfeeters R F.Metabolism of lactic acid in fermented cucumbers by Lactobacillus buchneri and related species,potential spoilage organisms in reduced salt fermentations[J].Food Microbiology,2013,35(2):129-135.
[26]Franco W,Pérez-díaz I M.Role of selected oxidative yeasts and bacteria in cucumber secondary fermentation associated with spoilage of the fermented fruit[J].Food Microbiology,2012,32(2):338-344.
[27]Laitila A,Sarlin T,Raulio M,et al.Yeasts in malting,with special emphasis on Wickerhamomyces anomalus(synonym Pichia anomala)[J].Antonie Van Leeuwenhoek,2011,99(1):75-84.
[28]张腾.发酵蔬菜微生物群落变化及与亚硝酸盐的关系[D].太原:山西大学,2013.
[29]Krishnan P,Bhat R,Kush A,et al.Isolation and functiona characterization of bacterial endophytes from Carica papaya fruits[J].Journal of Applied Microbiology,2012,113(2):308-317.
[30]Rao Y,Qian Y,She X,et al.Pellicle formation,microbia succession and lactic acid utilisation during the aerobic deteriorating process of Sichuan pickle[J].International Journal of Food Science and Technology,2017,53(3).
[31]Zogaj X,Bokranz W,Nimtz M,et al.Production of cellulose and curli fimbriae by members of the family Enterobacteriaceae isolated from the human gastrointestinal tract[J].Infection and Immunity,2003,71(7):4151-4158.
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