摘要
本文以大豆为原料,利用毛霉721-3和纳豆菌双菌组合发酵生产细菌型豆豉。通过化学分析、感官评价、仪器分析技术等手段对发酵过程中影响两种微生物的各种因素分别进行了探索研究,并通过正交实验初步确定了最佳发酵工艺。其研究的主要结论如下:
1、毛霉721-3分泌的酶系中蛋白酶和纤维素酶较多,其中蛋白酶主要为中性蛋白酶;其制种曲的最佳工艺为:调整种曲培养基pH至7,在25℃恒温培养72小时,即可制得酶活较高,孢子较多的优质种曲;
2、单一毛霉最佳的制曲工艺为:大豆浸泡至含水量45%左右;在0.1MPa下高压蒸汽蒸煮30min,冷却后接种3%湿豆重的优良种曲,25℃恒温培养48h,期间喷洒pH7的溶液以维持培养期间的相对湿度,即可得到蛋白酶活力较高的豉曲。
4、纳豆生产的最佳工艺条件为:水与大豆的比例为1:3,20℃浸泡10h,虑干后0.1MPa下高压蒸汽蒸煮30min;接种3%种龄为20h的种子液; 30℃下恒温发酵24h;发酵过程添加糯米粉8%、NaCll%和蔗糖4%有利于纳豆菌的生长,并且最终产品风味更佳;纳豆的保质期为4℃下15d。
5、在单菌发酵实验的基础上,将毛霉和纳豆菌进行复合生产大豆发酵细菌型豆豉。实验发现双菌发酵的最佳工艺为:毛霉和纳豆菌分别制种曲后以1:1的比例同时接种制作豆豉曲,接种量各为蒸煮后湿豆重的3%,豉曲pH调为7,加入糯米粉8%、NaCl1%和蔗糖4%,30℃下发酵18h,4℃后熟24h即可得纳豆激酶活力较强,氨基态含量高,但没有氨臭味的豆制品。
6、双菌发酵的工艺流程为毛霉和纳豆菌菌种→种曲↓大豆→筛选→浸泡→沥干→蒸煮→冷却→接种→发酵→后熟↑糯米粉、NaCl、蔗糖
This paper is about fermenting bean products by Dixenie consisting of Mucor 721-3 and Bacillus natto, as well as soybeans as the raw material. The separate exploration and research on all the factors of the effect of both microorganisms by means of chemical analysis, sense evaluation and instrumental analysis and the Orthogonal experiment basically confirmed the best ferment technology. The main conclusions of the research are as follow:
1. The protease and cellulase in enzyme that is secreted by Mucor 721-3 are much more, and protease is mainly neutral protease. The best technology of production of hybrid seeds is, adjusting the hybrid seeds pH to 7, cultivating them for 72 hours in constant temperature of 25℃, and we can get hybrid seeds that contains more active enzyme and more spore.
2. The optimum technology of production of hybrid seeds by single Mucor is soaking the soybeans to the moisture content of about 45%, stewing them for 30 minutes under the high pressure steam of O.1MPa, inoculating the good hybrid seeds of 3% of wet soybeans after cooling, and then cultivating them for 48 hours in constant temperature of 25℃, and meanwhile sprinkling the solution of pH in order to keep the relative humidity during this period, and then we can get the fermented soya beans with more active protease.
3. The optimum technological conditions of production of natto are, the proportion of water and soybean is 1 to 3; soaking the soybeans for 10 hours in constant temperature of 20℃, stewing them for 30 minutes under the high pressure steam of 0.1MPa after filtering, inoculating 3% of seed liquor aged 20 hours; then fermenting them for 24 hours in constant temperature of 30℃, during which 8% of glutinous rice flour,1% of NaCl and 4% of cane sugar must be added in order to be good for the growth of the Bacillus natto and make the production taste better. The quality guarantee period of natto is 14 days in the temperature of 4℃.
4. Ferment by complex production of Mucor and Bacillus natto based on the experiment of Zymomonas mobilis. According to the experiment, the optimum technology of Double bacteria fermentation is, inoculating and producing the hybrid seeds after producing the hybrid seeds with Mucor and Bacillus natto separately, and the amount of the inoculation is 3% of wet soybeans after stewing, the pH of hybrid seeds is 7, adding 8% of glutinous rice flour,1% of NaCl and 4% of cane sugar and fermenting 18 hours in the temperature of 30℃, after-ripening for 24 hours in the temperature of 4℃, and we can get soybean product with more active natto kinase and richer nutrition
5. The process of the technique of Double bacteria fermentation is as follow:
Mucor and Bacillus natto slope strain→mouldstarter↓Soybean→sifting→soaking→Draining→stewing→cooling→inoculating→fermenting→after-ripeni ng↑glutinous rice flour, NaCl, cane sugar
引文
[1]孙森,宋俊梅.豆豉、纳豆及天培的研究进展[J].中国调味品,2008,3:29-33.
[2]房翠兰.豆豉加工过程中蛋白质和膳食纤维生物学变化的研究[硕十学位论文].重庆:西南大学,2007.
[3]夏岩石,夏延斌.豆豉的营养与保健功能.中国食物与营养.2005,2:48-50.
[4]穆慧林,李里特.豆豉的保健功能及其开发价值[J].农产品加工.学刊,2008(10):30-32.
[5]宋永生,张炳文.日本纳豆与中国豆豉营养功能成分的研究进展[J].中国调味品,2004(12):6-9.
[6]李里特,张建华,李再贵等.纳豆、天培与豆豉的比较[J].中国调味品,2003,(2):3-8.
[7]Hideo Esaki, Hiromichi Onozaki, Shunro Kawakishi et al. Antioxidant activity an d isolation from soybeans fermented with Aspergillus spp[J]. J Agric Food Chem,1997,45:2020-2022.
[8]李幼筠,甘萍等.毛霉型豆豉生产菌种的研究[J].中国酿造,1989;(8):29-31.
[9]苗兰兰,等.永川豆豉中高产蛋白酶菌株的筛选鉴定及液体培养条件的研究[J].农产品加工·学刊,2009,5(5):66-90.
[10]李祥.细菌型豆豉生产的研究[J].中国调味品,1999,(10):14-17.
[11]孙森,宋俊梅.豆豉后发酵过程中微生物菌相的变化[J].开发应用及中国食品添加剂,2008(2):139-143.
[12]钱家亮,等.豆豉天然制曲过程的动态研究[J].食品与发酵工业,2008,34(2):58-60.
[13]彭勇,张义正.豆豉溶栓酶产生菌的筛选及其酶学性质的初步研究[J].高技术通讯,2002(2):30-34.
[14]袁贵英.河南臭豆豉发酵微生物的分离鉴定[J].江苏调味副食品,2007,24(2):28-36.
[15]鄯晋晓.细菌型豆豉的研究现状及发展前景[J].中国酿造,2007,3:1-4.
[16]秦礼康.陈窖豆豉粑传统工艺剖析及优势菌群鉴定[J].食品科学,2006,27:118-123.
[17]吴龙英.产酱香芽孢杆菌在豆豉发酵上的应用[J].山地农业生物学,2000(3):237-238.
[18]贾东旭.细菌型豆豉发酵芽孢杆菌的筛选与鉴定[J].食品科学,2009,30(5):217-221.
[19]田亚红.豆豉中纳豆芽孢杆菌的筛选及其发酵工艺的研究[J].中国酿造,2009,4(205):45-48.
[20]Perez, Marlen, Study of microbial contamination of tempeh, J. Alimentaria,2003,40:129-132.
[21]B.Ashenafi, M.and M.Busse, Growth of Bacillus cereus in fermenting tempeh made from various beans and its inhibition by Lactobacillus plantarum, J.Appl.Bacteriol,1991.70:329-333.
[22JP.K. Sarkar,B.Hasenack,M.J.R.Nout, Diversity and functionality of Bacillus and related genera isolated from spontaneously fermented soybeans (Indian Kinema) and locust beans (African Soumbala), J. Food Microbiology 2002,77:175-186.
[23]K. Jeyaram,W. Mohendro Singh, Molecular identification of dominant microflora associated with 'Hawaijar'— A traditional fermented soybean (Glycine max (L.)) food of Manipur, India, J. Food Microbiology,2008,122:259-26
[24]Jennifer Jennessena,Johan Schnurera, Morphological characteristics of sporangiospores of the tempe fungus Rhizopus oligosporus differentiate it from other taxa of the R. microsporus group, J.Mycological Research,2008,112:547-563.
[25]Ogawa, Yoshio, An original habitat of tempeh molds, J. Mycoscience,2004,45:271-276
[26]P.K.Sarkar,B.Hasenack,M.J.R.Nout, Diversity and functionality of Bacillus and related genera isolated from spontaneously fermented soybeans (Indian Kinema) and locust beans (African Soumbala), J. Food Microbiology 2002,77:175-186.
[27]张建华.曲霉型豆豉曲中微生物分布[J].中国调味品,2006,9(9):15-20.
[28]蒋立文,周传云,李宗军.传统发酵大豆制品的质量与安全控制探讨[J].中国酿造,2006(1):1-3.
[29]秦恩华.土家豆豉中微生物的初步研究[J].2007,01:30-39.
[30]庞庆芳,张炳文.中国传统大豆发酵食品—豆豉功能成分的研究进展[J].食品研究与开发,2006,27(2):185-198.
[31]蔡位辉.豆豉生产新工艺的研究[D].无锡轻工大学硕士学位论文,1997
[32]杜木英,毛霉型豆豉生物速成发酵技术的研究[硕士学位论文].重庆:西南大学,2001.
[33]夏岩石,夏延斌.毛霉豆豉前发酵条件优化的研究[J].湖南科技学院学报,2010,4:83-86.
[34]谭周进,周传云.原料对纳豆品质的影响[J].食品科学,2003,24(1):87-90.
[35]李华,李铎.细菌型豆豉纯种发酵工艺优化[J].中国粮油学报,2009,24(2):50-54.
[36]奚锐华,齐凤兰.纳豆生产工艺的优化[J].食品工业科技,2004,3:78-80
[37]祖国仁,孔繁东.纳豆菌固体发酵条件及产品成分分析[J].食品工业科技,2006,12:122-124.
[38]孙成行,眸光庆.豆豉混合菌种制曲工艺的研究[J].中国调味品,2007,3:43-46.
[39]杜海清,焦宇知.豆豉混种制曲工艺优化[J].中国酿造,2008,16:72-74.
[40]代丽娇.缩短豆豉生产周期的研究[D].山东轻工业学院硕十学位论文,2007.
[41]MariaPilarMarti,etal.Solid-PhasemicroextractionandgaschromatographyOlfactometryanalysisofs uccessivelydi-lutedsamples.Anewapproachofthearomaextractdilutionanalysisappliedtothecharacteriz ationofwinearoma[J].AgricFoodChem,2003,51:7861-7865.
[42]Rimantas P Venskutonis,et al.Use of GC-headspace and"electronicnose" forthedetection of volatile compounds fromglucose-glycineMaillardreaction[J].FoodControl,2002,13:13-21.
[43]Leejeerajumnean,etal.VolatilecompoundsinBacillus-fer-mentedsoybeans[J].SciFoodAgric,2001, 81(5):525-529.
[44]余爱农,杨春海,谭志斗,等.细菌型豆豉香气成分的研究[J].食品科学,2002,23(12):98-100.
[45]秦礼康,丁霄霖.传统陈窖豆豉粑和霉菌型豆豉挥发性风味化合物研究[J].食品科学,2005,26(8):275-280.
[46]张惟广,张惠玲.酿造工艺学.成都:成都科技大学出版社,1998.
[47]沈萍,范秀荣.微生物学实验第3版[M].北京:高等教育出版社,1999.6(2006重印)
[48]夏岩石,夏延斌.毛霉豆豉前发酵条件优化的研究[J].湖南科技学院学报,2010,4:83-86.
[49]杨军,陈九武.曲霉型豆豉生产工艺的研究.中国调味品,1998,(4):17-18,23
[50]北京大学生物化学教研室.生物化学实验指导.北京:人民教育出版社,1979
[51]范秀容.微生物学实验.高等教育出版社,1989
[52]宋国安.功能保健食品纳豆的研究与开发[J].武汉工业学院学报,2002(1):40-42
[53]Sumi H,Hamada H,Tsushima H,et al. A novel fibrinolytic enzyme(nattokinase) in the vegetable cheese natto, a typical and popular soybean food in the Japanese diet [J]. Experientia, 1987,43(10):1110-1111.
[54]钟青萍,佘世望,梁胜媛.纳豆菌的抗菌作用及其培养基的优化[J].食品工业科技,2001,22(5):20-22.
[55]中华人民共和国卫生部药典委员会编.中华人民共和国药典(二部)[M].北京:化学工北出版社,1995.320~419.
[56]张菁,朱镇华等.尿激酶原活性测定方法研究[J].南京大学学报(自然科学),2001,37(4):416-419.
[57]大连轻工业学院,华南理工大学,郑州轻工业学院等合编.食品分析[M].北京:中国轻工业出版社,1944,10.221.
[58]高俊风编.植物生理学实验技术[M].西安:西北农业大学出版,1992.135-138.
[59]张惟杰.复合多糖生化研究技术[M].上海:上海科学技术出版社,1987.6-7.
[60]周建平,郭华.纳豆黏液成分分析[J]食品工业科技,2003,24(2):32-34.