CN110101076A - 一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法 - Google Patents
一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法 Download PDFInfo
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- CN110101076A CN110101076A CN201910288573.1A CN201910288573A CN110101076A CN 110101076 A CN110101076 A CN 110101076A CN 201910288573 A CN201910288573 A CN 201910288573A CN 110101076 A CN110101076 A CN 110101076A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Landscapes
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- Mycology (AREA)
- Health & Medical Sciences (AREA)
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- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Preparation Of Fruits And Vegetables (AREA)
Abstract
本发明公开了一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法,属于发酵食品加工技术领域。首先制备曲霉、乳酸菌、双歧杆菌和酵母种子;其次,基于中药配伍理论确定了以牛蒡为“君”及以豆类、莲藕和红枣为“辅”的原料搭配;再次,采用分段发酵和不同菌种分开酿造技术进行发酵,达到大量增殖益生菌及催化低活性糖苷型黄酮转变成高活性黄酮苷元的目的;最后,结合运用酵素稳态化和真空充氮技术使酵素中的活性益生菌和黄酮苷元保存。按照本发明方法生产的牛蒡酵素储存90d时活性黄酮苷元和益生菌含量分别达到100‑350μg/mL、活性益生菌数达到1‑10×109CFU/mL。
Description
技术领域
本发明涉及一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法,属于发酵食品加工技术领域。本发明以牛蒡、豆类(大豆、绿豆、赤豆)、莲藕和红枣为主要原料,以曲霉、乳酸菌、双歧杆菌和酵母为发酵菌种,采用分段酿造方式,结合利用稳态化处理和真空充氮保藏技术制备的一种健康饮品。
背景技术
牛蒡(Arctium lappa L.),又名大力子(《本草纲目》)、白肌人参,亦称“东洋萝卜”、“黑萝卜”,别称“恶实”,是菊科(composite)牛蒡属直根系二年生大型草本植物。牛蒡原产于我国,以野生为主,公元940年前后传入日本,被培育出多个优良品种,成为日本人喜食的蔬菜(贺菊萍,牛蒡中活性物质的提取、分离纯化及其生理功能的研究进展[J],食品工业科技,2012,33(10):407-411.)。目前牛蒡的种植主要分布于中国、西欧、克什米尔地区、欧洲等地。中国牛蒡的产地主要分布于江苏省(徐州丰县、沛县)和山东省(苍山)。牛蒡保存过程中易水分流失而导致其木质化、变硬和风味丧失,进而导致牛蒡品质和商品价值下降,农民丰产不丰收(王乃馨等,复合牛蒡果蔬酵素饮料配方优化研究[J],现代食品,2018,(17):137-144.)。牛蒡深加工技术不但可以提高牛蒡的附加值,也可以延长产品的保藏期,是解决上述牛蒡产业问题的重要技术。
牛蒡能清除体内自由基和改善体循环,尤其对糖尿病、性机能减退、肥胖症、风湿、类风湿、解肝毒、便秘、内外痔等病症有明显效果(贺菊萍,牛蒡中活性物质的提取、分离纯化及其生理功能的研究进展[J],《食品工业科技》,2012,33(10):407-411.)。牛蒡富含黄酮类物质是其具有上述功能的关键物质基础之一(蒋晓文等,牛蒡根中黄酮苷类化学成分及其抗氧化活性构效关系的研究[J],《中草药》,2016,47(5):726-731.)。大量的研究已经证实豆类、莲藕和红枣具有抗癌、抗氧化、抗感染、抗突变和抗动脉粥样硬化等生理活性,而其中的黄酮类物质是重要的功能因子(Lee J.H.,et al.Comparisons of nutritionalconstituents in soybeans during solid state fermentation times and screeningfor their glucosidase enzymes and antioxidant properties[J],Food Chemistry,2019,272,362-371.;Ji L.,et al.,In vivo antioxidant properties of lotus rootand cucumber:Apilot comparative study in aged subjects[J],The Journal ofNutrition,Health and Aging,2015,19(7):765-770.;周晓凤等.不同枣品种果实中功能性成分含量的比较研究[J],《食品工业科技》,2018,(12):296-300)。牛蒡、豆类、莲藕和红枣均为营养和安全的食材,依据中药配伍中的“君臣佐使”理论和“四气、五味和升降沉浮”学说,以牛蒡根为“君”并辅以豆类、莲藕和红枣为原料开发成具有一定生理功能的新型饮料具有很好的市场前景。
酵素是指以大豆、小麦、米胚芽和各种蔬菜水果等为原料,用乳酸菌、酵母菌和曲霉等为菌种发酵而成的食品。酵素中主要有益成分来源于原料本身和/或者微生物代谢产物和/或活性微生物本身,主要含有酶(蛋白酶、脂肪酶、淀粉酶、超氧化物歧化酶)、黄酮、多酚、维生素、矿物质和次生代谢产物等,具有润肠通便、抗氧化、抑菌消炎、消化分解、美容养颜等功能,并能改善宿醉、头疼等现象,还能有效增强黏膜系统的免疫功能(费爽雯等,几种市售酵素中活性物质的比较[J],南昌大学学报(工科版),2017,39(1):27-31.;胡学智,“酵素”和酶的辨异—论酶和“酵素”制品的保健功能[J],工业微生物,2016,46(2):60-68.)。但酵素中的酶类和活性微生物在酵素储存和进入人体消化系统时(特别是胃)由于环境温度、氧气的消耗和pH的剧烈变化容易失活,因此酵素的有益效果将显著降低(唐超等,食用酵素的研究进展[J],生物加工过程,2018,16(3):95-103.)。此外,牛蒡、豆类和红枣黄酮多以糖苷形式存在,研究已经证实以糖苷形式存在的黄酮类化合物活性远低于以苷元形式存在的黄酮。而这些低活性的糖苷型黄酮能够通过微生物发酵作用转化为具有更高活性的黄酮苷元,从而提高其生理活性(胡学智,“酵素”和酶的辨异—论酶和“酵素”制品的保健功能[J],工业微生物,2016,46(2):60-68.)。
本发明基于中药配伍理论,在大量实验基础上以牛蒡为关键原料、以曲霉、乳酸菌、双歧杆菌和酵母为发酵菌种结合生物活性物质稳态化和真空充氮技术开发出一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法。该方法能够显著提高牛蒡酵素黄酮苷元和活性益生菌数量,在保证牛蒡酵素良好风味和体态稳定的情况下,保存90d时黄酮苷元含量达到100-350μg/mL、活性益生菌数达到1-10×109CFU/mL。因此本发明具有重要的应用价值和经济意义。
发明内容
本发明的目的是为了开发一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法。具体地,本发明提供了一种利用牛蒡、豆类、莲藕和大枣为原料、利用曲霉、乳酸菌、双歧杆菌和酵母发酵作用生产牛蒡酵素的方法。
一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法,按照下述步骤进行:
(1)曲霉种子制备:采用米曲霉(Aspergillus oryzae)3.042和/或酱油曲霉(Aspergillus sojae)为菌种,麸皮经润水(麸皮:水=1:0.7,w/w)、121℃蒸煮10min、冷却(40℃)、接种(1-10×106个孢子/g)后于28℃下在曲房培养至种曲表面黄绿色(期间温度上升至35℃时翻曲2次),制备好的麸皮种曲待用;
(2)乳酸菌种子制备:以德氏乳杆菌保加利亚亚种(Lactobacillus Delbrueckiisubsp.bulgaricus)和/或植物乳杆菌(Lactobacillus plantarum)和/或嗜热链球菌(Streptococcus thermophilus)和/或保加利亚乳杆菌(Lactobacillus bulgaricus)为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(1-10×106CFU/g)后于25-43℃摇床中(180rpm)培养8-24h,得乳酸菌种子液;
(3)双歧杆菌种子制备:以青春双岐杆菌(Bifidobacterium adolescentis)和/或乳双岐杆菌(Bifidobacterium lactis)和/或婴儿双岐杆菌(Bifidobacterium infantis)为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁(提供部分维生素和矿物质):水=20:10:7:3:0.5:8:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(1-10×106CFU/g)后于25-40℃摇床中(180rpm)培养10-20h,得双歧杆菌种子液;
(4)酵母菌种子的制备:以酿酒酵母(Saccharomyces cerevisiae)和/或卡氏酵母(Saccharomyces carlsbergensis)为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种(1-10×106CFU/g)后于20-38℃摇床中(180rpm)培养20-48h,得酵母菌种子液;
(5)原料配伍与处理:一种富含黄酮苷元和活性益生菌牛蒡酵素原料,配伍为牛蒡:大豆:绿豆:赤豆:莲藕:红枣=100:20-200:0-200:0-200:5-50:5-50。豆类(大豆、绿豆、赤豆)清洗后加水浸泡5-12h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料(牛蒡、豆类、莲藕和红枣):水=1:7-20的比例混合后磨浆,混合浆液于100-131℃灭菌30s-30mi后待用;
(6)一次发酵(曲霉发酵):将混合浆液转至无菌发酵罐,待混合浆液降温至40℃时接入曲霉种子,使混合浆液中曲霉孢子数达到1-10×106CFU/mL、搅拌速度50-200rpm、通氧量10-60L/h、发酵温度20-40℃、发酵时间10-120h;
(7)过滤及灭菌:发酵结束后板框过滤机过滤,滤液于100-131℃下灭菌15s-15min后冷却至35℃待用;
(8)二次发酵(细菌和酵母菌发酵):取20-40%步骤(7)灭菌液接种1-10%乳酸菌种子液,于25-40℃培养5-20h;取20-40%步骤(7)灭菌液接种1-10%双歧杆菌种子液,于25-40℃培养5-20h;取剩余20-60%步骤(7)灭菌液接种1-10%酵母菌种子液,于20-38℃培养10-30h;发酵结束后将三者混合均匀;
(9)调配:按照混合发酵液:白砂糖:蜂蜜:低聚果糖=100:0-10:0-5:0-5进行调配(w/w),使终产品具有宜人口感;
(10)稳态化处理及包装:将海藻糖、羧甲基纤维素钠、黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到0.3-5%、0.1-2%和0.01-1%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装;
(11)检验:利用高效液相法检测终产品中黄酮苷元含量,黄酮苷元含量达到100-350μg/mL;平板法培养检测终产品中活性益生菌数,活性益生菌数达到1-10×109CFU/mL;产品不得检出大肠杆菌;符合上述指标即为合格产品。
本发明与现有酵素生产方法相比,本发明方法具有如下优点和有益效果:
本发明方法生产的酵素均以常规食材为原料、发酵菌种均为GRAS级微生物,生产过程不使用和添加化学试剂(包括防腐剂),保证了原料的绿色和安全。生产过程不采用超高温(>160℃)和长时间(超过2h)加热方式灭菌避免产品产生有害物质。得益于原料的合理配伍、有益微生物的大量繁殖、糖苷型黄酮高效转化为黄酮苷元及合理的稳态化处理技术和真空充氮技术的综合利用,终产品黄酮苷元和活性益生菌含量在保藏90d时仍高于100μg/mL和1×109CFU/mL。终产品符合消费者对绿色、安全、健康食品的诉求,产品附加值显著提高。因此,本发明具有重要的应用价值和经济价值。
具体实施方式
下面结合实例对本发明的具体实施方式作进一步说明,但本发明的实施和保护范围不限于此。
本发明所涉及的黄酮苷元测定方法参照Lee等方法(Lee J.H.,etal.Comparisons of nutritional constituents in soybeans during solid statefermentation times and screening for their glucosidase enzymes andantioxidant properties[J],Food Chemistry,2019,272,362-371.),菌落总数和大肠菌群数测定分别参照GB 4789.2-2016和GB 4789.3-2016方法,糖含量测定参照GB/T 5009.7-2008方法。
实施例1一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法—以牛蒡、大豆、绿豆、赤豆、莲藕和红枣为原料
曲霉种子的制备采用米曲霉(Aspergillus oryzae)3.042和酱油曲霉(Aspergillus sojae)为菌种,麸皮经润水(麸皮:水=1:0.7)、蒸煮(121℃/10min)、冷却(40℃)、接种(1×107个孢子/g)后于28℃下在曲房培养至种曲表面黄绿色(期间温度上升至35℃时翻曲2次),制备好的麸皮种子待用。乳酸菌种子制备以德氏乳杆菌保加利亚亚种(Delbrueckii subsp.bulgaricus)、植物乳杆菌(Lactobacillus plantarum)、嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus)为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000(重量比)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(1×107CFU/g)后于43℃摇床中(180rpm)培养24h,得乳酸菌种子液。双歧杆菌种子制备以青春双岐杆菌(Bifidobacterium adolescentis)、乳双岐杆菌(Bifidobacterium lactis)和婴儿双岐杆菌(Bifidobacterium infantis)为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁:水=20:10:7:3:0.5:8:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(1×107CFU/g)后于40℃摇床中(180rpm)培养20h,得双歧杆菌种子液。酵母菌种子的制备以酿酒酵母(Saccharomyces cerevisiae)和卡氏酵母(Saccharomyces carlsbergensis)为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种(1×107CFU/g)后于38℃摇床中(180rpm)培养48h,得酵母菌种子液。牛蒡酵素原料配伍为牛蒡:大豆:绿豆:赤豆:莲藕:红枣=100:200:200:200:50:50(w/w)。豆类清洗后加水浸泡12h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料(牛蒡、豆类、莲藕和红枣):水=1:7的比例混合后磨浆,混合浆液于131℃条件下灭菌30s后待用。将混合浆液转至无菌发酵罐,待混合浆液降温至40℃时接入曲霉种子,使混合浆液中曲霉孢子数达到1×107CFU/mL、搅拌速度200rpm、通氧量60L/h、发酵温度40℃、发酵时间120h。发酵结束后用板框过滤机过滤,滤液于100℃条件下灭菌15min后冷却至35℃待用。取40%灭菌液接种10%乳酸菌种子液,于40℃培养20h;取40%灭菌液接种5%双歧杆菌种子液,于40℃培养20h;取剩余20%灭菌液接种1%酵母菌种子液,于38℃培养30h;发酵结束后将三者混合均匀。按照白砂糖:蜂蜜:低聚果糖=100:0:0:0进行调配(w/w)进行调配。将海藻糖、羧甲基纤维素钠、黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到0.3%、0.1%和0.01%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装。
采用本发明方法生产的牛蒡酵素与其他牛蒡酵素相比,本发明方法生产的牛蒡酵素酸爽可口、无异味,体态和色泽稳定。保存90d时终产品中活性成分和活性益生菌保持稳定,黄酮苷元含量达到350μg/mL、活性益生菌数达到10×109CFU/mL、未检出大肠杆菌。
实施例2一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法—以牛蒡、大豆、绿豆、莲藕和红枣为原料
曲霉种子的制备采用酱油曲霉(Aspergillus sojae)为菌种,麸皮经润水(麸皮:水=1:0.7)(w/w)、蒸煮(121℃/10min)、冷却(40℃)、接种(5×106孢子/g)后于28℃下在曲房培养至种曲表面黄绿色(期间温度上升至35℃时翻曲2次),制备好的麸皮种子待用。乳酸菌种子制备以德氏乳杆菌保加利亚亚种(Lactobacillus Delbrueckiisubsp.bulgaricus)和植物乳杆菌(Lactobacillus plantarum)为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(5×106CFU/g)后于34℃摇床中(180rpm)培养16h,得乳酸菌种子液。双歧杆菌种子制备以青春双岐杆菌(Bifidobacterium adolescentis)和乳双岐杆菌(Bifidobacterium lactis)为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁:水=20:10:7:3:0.5:8:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(5×106CFU/g)后于33℃摇床中(180rpm)培养15h,得双歧杆菌种子液。酵母菌种子的制备以酿酒酵母(Saccharomyces cerevisiae)为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种(5×106CFU/g)后于29℃摇床中(180rpm)培养34h,得酵母菌种子液。牛蒡酵素原料配伍为牛蒡:大豆:绿豆:莲藕:红枣=100:110:100:25:25(w/w)。豆类(大豆、绿豆)清洗后加水浸泡8h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料(牛蒡、豆类、莲藕和红枣):水=1:13的比例混合后磨浆,混合浆液于111℃灭菌15min后待用。将混合浆液转至无菌发酵罐,待混合浆液降温至40℃时接入曲霉种子,使混合浆液中曲霉孢子数达到5×106CFU/ml、搅拌速度125rpm、通氧量35L/h、发酵温度30℃、发酵时间65h。发酵结束后用板框过滤机过滤,滤液于110℃灭菌7.5min后冷却至35℃待用。取30%灭菌液接种5%乳酸菌种子液,于33℃培养12h;取30%灭菌液接种1%双歧杆菌种子液,于33℃培养12h;取剩余40%灭菌液接种5%酵母菌种子液,于29℃培养20h;发酵结束后将三者混合均匀。按照混合发酵液:白砂糖:蜂蜜:低聚果糖=100:5:2.5:2.5进行调配(w/w),使终产品具有宜人口感。将海藻糖、羧甲基纤维素钠、黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到2.6%、1%和0.5%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装。
采用本发明方法生产的牛蒡酵素与其他牛蒡酵素相比,本发明方法生产的牛蒡酵素酸甜协调、无异味,体态和色泽稳定。保存90d时终产品中活性成分和活性益生菌保持稳定,黄酮苷元含量达到250μg/mL、活性益生菌数达到6×109CFU/mL、未检出大肠杆菌。
实施例3一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法—以牛蒡、大豆、赤豆、莲藕和红枣为原料
曲霉种子的制备采用米曲霉(Aspergillus oryzae)为菌种,麸皮经润水(麸皮:水=1:0.7)(w/w)、蒸煮(121℃/10min)、冷却(40℃)、接种(2×106孢子/g)后于28℃下在曲房培养至种曲表面黄绿色(期间温度上升至35℃时翻曲2次),制备好的麸皮种子待用。乳酸菌种子制备以德氏乳杆菌保加利亚亚种(Lactobacillus Delbrueckii subsp.bulgaricus)和嗜热链球菌(Streptococcus thermophilus)为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(3.5×106CFU/g)后于30℃摇床中(180rpm)培养20h,得乳酸菌种子液。双歧杆菌种子制备以青春双岐杆菌(Bifidobacterium adolescentis)和婴儿双岐杆菌(Bifidobacteriuminfantis)为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁:水=20:10:7:3:0.5:8:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(7×106CFU/g)后于29℃摇床中(180rpm)培养17h,得双歧杆菌种子液。酵母菌种子的制备以酿酒酵母(Saccharomyces cerevisiae)为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种(3×106CFU/g)后于29℃摇床中(180rpm)培养40h,得酵母菌种子液。牛蒡酵素原料配伍为牛蒡:大豆:赤豆:莲藕:红枣=100:90:90:25:25(w/w)。豆类(大豆、赤豆)清洗后加水浸泡10h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料(牛蒡、豆类、莲藕和红枣):水=1:12的比例混合后磨浆,混合浆液于115℃下灭菌20min后待用。将混合浆液转至无菌发酵罐,待混合浆液降温至40℃时接入曲霉种子,使混合浆液中曲霉孢子数达到3×106CFU/ml、搅拌速度120rpm、通氧量38L/h、发酵温度32℃、发酵时间60h。发酵结束后用板框过滤机过滤,滤液于115℃灭菌9min后冷却至35℃待用。取25%灭菌液接种4%乳酸菌种子液,于30℃培养15h;取35%灭菌液接种3%双歧杆菌种子液,于32℃培养13h;取剩余40%灭菌液接种4%酵母菌种子液,于30℃培养25h;发酵结束后将三者混合均匀。按照混合发酵液:白砂糖:蜂蜜:低聚果糖=100:3:1.5:1.5进行调配(w/w),使终产品具有宜人口感。将海藻糖、羧甲基纤维素钠、黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到2.0%、0.8%和0.6%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装。
采用本发明方法生产的牛蒡酵素与其他牛蒡酵素相比,本发明方法生产的牛蒡酵素酸甜协调、无异味,体态和色泽稳定。保存90d时终产品中活性成分和活性益生菌保持稳定,黄酮苷元含量达到300μg/mL、活性益生菌数达到8×109CFU/mL、未检出大肠杆菌。
实施例4一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法—以牛蒡、大豆、莲藕和红枣为原料
曲霉种子的制备采用米曲霉(Aspergillus oryzae)3.042为菌种,麸皮经润水(麸皮:水=1:0.7)、蒸煮(121℃/10min)、冷却(40℃)、接种(1×106个孢子/g)后于28℃下在曲房培养至种曲表面黄绿色(期间温度上升至35℃时翻曲2次),制备好的麸皮种子待用。乳酸菌种子制备以德氏乳杆菌保加利亚亚种(Lactobillus Delbrueckii subsp.bulgaricus)为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时各菌种接种(1×106CFU/g)后于25℃摇床中(180rpm)培养8h,得乳酸菌种子液。双歧杆菌种子制备以青春双岐杆菌(Bifidobacterium adolescentis)为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁:水=20:10:7:3:0.5:8:1000(w/w)的比例混合后于113℃灭菌20min,降温至40℃时接种(1×106CFU/g)后于25℃摇床中(180rpm)培养10h,得双歧杆菌种子液。酵母菌种子的制备以酿酒酵母(Saccharomycescerevisiae)为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种(1×106CFU/g)后于20℃摇床中(180rpm)培养20h,得酵母菌种子液。牛蒡酵素原料配伍为牛蒡:大豆:莲藕:红枣=100:20:5:5(w/w)。大豆清洗后加水浸泡5h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料(牛蒡、大豆、莲藕和红枣):水=1:20的比例混合后磨浆,混合浆液于100℃下灭菌30min后待用。将混合浆液转至无菌发酵罐,待混合浆液降温至40℃时接入曲霉种子,使混合浆液中曲霉孢子数达到1×106CFU/ml、搅拌速度50rpm、通氧量10L/h、发酵温度20℃、发酵时间10h。发酵结束后用板框过滤机过滤,滤液于131℃下灭菌/15s后冷却至35℃待用。取20%灭菌液接种1%乳酸菌种子液,于25℃培养5h;取20%灭菌液接种10%双歧杆菌种子液,于25℃培养5h;取剩余60%灭菌液接种10%酵母菌种子液,于20℃培养10h;发酵结束后将三者混合均匀。按照混合发酵液:白砂糖:蜂蜜:低聚果糖=100:10:5:5进行调配(w/w),使终产品具有宜人口感。将海藻糖、羧甲基纤维素钠、黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到5%、2%和1%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装。
采用本发明方法生产的牛蒡酵素与其他牛蒡酵素相比,本发明方法生产的牛蒡酵素甜酸突出、无异味,体态和色泽稳定。保存90d时终产品中活性成分和活性益生菌保持稳定,黄酮苷元含量达到100μg/mL、活性益生菌数达到1×109CFU/mL、未检出大肠杆菌。
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做出各种改动与修饰,因此本发明的保护范围应该以权利要求书所界定范围为准。
Claims (8)
1.一种富含黄酮苷元和活性益生菌牛蒡酵素的生产方法,其特征在于按照下述步骤进行:
(1)曲霉种子制备:采用米曲霉(Aspergillus oryzae)和/或酱油曲霉(Aspergillussojae)为菌种,麸皮经润水,麸皮与水的重量比为1:0.7、121℃蒸煮10min、冷却至40℃、接种后于28℃下在曲房培养至种曲表面黄绿色,期间温度上升至35℃时翻曲2次,制备好的麸皮种曲待用,其中接种曲霉孢子数为1-10×106个孢子/g;
(2)乳酸菌种子制备:以德氏乳杆菌保加利亚亚种(Lactobacillus Delbrueckiisubsp.bulgaricus)和/或植物乳杆菌(Lactobacillus plantarum)和/或嗜热链球菌(Streptococcus thermophilus)和/或保加利亚乳杆菌(Lactobacillus bulgaricus)为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000的重量比例混合后于113℃灭菌20min,降温至40℃时各菌种接种后于25-43℃摇床中,180rpm培养8-24h,得乳酸菌种子液,其中,接种用乳酸菌的活菌数为1-10×106CFU/g;
(3)双歧杆菌种子制备:以青春双岐杆菌(Bifidobacteriumadolescentis)和/或乳双岐杆菌(Bifidobacterium lactis)和/或婴儿双岐杆菌(Bifidobacteriuminfantis)为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁:水=20:10:7:3:0.5:8:1000的重量比例混合后于113℃灭菌20min,降温至40℃时,各菌种接种后于25-40℃摇床中,180rpm培养10-20h,得双歧杆菌种子液,其中,接种用双歧杆菌的活菌数为1-10×106CFU/g;
(4)酵母菌种子的制备:以酿酒酵母(Saccharomyces cerevisiae)和/或卡氏酵母(Saccharomyces carlsbergensis)为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种后于20-38℃摇床中180rpm培养20-48h,得酵母菌种子液,其中,接种用酵母的活菌数为1-10×106CFU/g;
(5)原料配伍与处理:一种富含黄酮苷元和活性益生菌牛蒡酵素原料配伍时重量比为:牛蒡:豆类:莲藕:红枣=100:20-200:5-50:5-50,其中豆类包括大豆和/或绿豆和/或赤豆,其重量比为大豆:绿豆:赤豆=100:0-200:0-200;豆类清洗后加水浸泡5-12h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料:水=1:7-20的比例混合后磨浆,混合浆液于100-131℃条件下灭菌30s-30min后待用;
(6)曲霉一次发酵:将混合浆液转至无菌发酵罐,待混合浆液降温至40℃时接入曲霉种子,使混合浆液中曲霉孢子数达到1-10×106CFU/mL、搅拌速度50-200rpm、通氧量10-60L/h、发酵温度20-40℃、发酵时间10-120h;
(7)过滤及灭菌:发酵结束后板框过滤机过滤,滤液于100-131℃条件下灭菌15s-15min后;冷却至35℃待用;
(8)细菌和酵母菌二次发酵:取20-40%步骤(7)灭菌液接种1-10%乳酸菌种子液,于25-40℃培养5-20h;取20-40%步骤(7)灭菌液液接种1-10%双歧杆菌种子液,于25-40℃培养5-20h;取剩余20-60%步骤(7)灭菌液接种1-10%酵母菌种子液,于20-38℃培养10-30h;发酵结束后将三者混合均匀;
(9)调配:混合发酵液与一定比例的白砂糖和/或蜂蜜和/或低聚糖进行调配,使终产品具有宜人口感,其重量比为白砂糖:蜂蜜:低聚果糖=100:0-50:0-50;
(10)稳态化处理及包装:将海藻糖、羧甲基纤维素钠、黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到0.3-5%、0.1-2%和0.01-1%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装;
(11)检验:终产品黄酮苷元含量含量达到100-350μg/mL;活性益生菌数达到1-10×109CFU/mL;不得检出大肠杆菌;符合上述指标即为合格产品。
2.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(2)中菌种为德氏乳杆菌保加利亚亚种(Delbrueckii subsp.bulgaricus)和/或植物乳杆菌(Lactobacillus plantarum)和/或嗜热链球菌(Streptococcusthermophilus)和/或保加利亚乳杆菌(Lactobacillus bulgaricus)中的一种或者一种以上为菌种,按照脱脂奶粉:酵母粉:葡萄糖:水=120:10:10:1000的重量比混合后于113℃灭菌20min,降温至40℃时各菌种接种后于30-40℃摇床中,180rpm培养10-18h,得乳酸菌种子液,其中接种的乳酸菌的活菌数为1-5×106CFU/g。
3.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(3)中以青春双岐杆菌(Bifidobacterium adolescentis)和/或乳双岐杆菌(Bifidobacterium lactis)和/或婴儿双岐杆菌(Bifidobacterium infantis)中的一种或二种或者三种为菌种,按照蔗糖:酪蛋白胨:酵母浸膏:低聚木糖:磷酸二氢钾:甘蓝汁:水=20:10:7:3:0.5:8:1000的重量比混合后于113℃灭菌20min,降温至40℃时各菌种接种后于30-40℃摇床中,180rpm培养10-16h,得双歧杆菌种子液,其中,接种的双歧杆菌的活菌数为1-5×106CFU/g。
4.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(4)中以酿酒酵母(Saccharomyces cerevisiae)和/或卡氏酵母(Saccharomyces carlsbergensis)中的一种或者二种为种子,5°Bé麦芽汁121℃灭菌20min降温至35℃时各菌种接种后于20-30℃摇床中,180rpm培养30-36h,得酵母菌种子液,其中,接种的酵母菌的活菌数为1-5×106CFU/g。
5.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(5)原料配伍时其重量比为牛蒡:豆类:莲藕:红枣=100:50-100:20-40:10-30,其中豆类包括大豆和/或绿豆和/或赤豆,其重量比为大豆:绿豆:赤豆=100:0-100:0-100;豆类清洗后加水浸泡8-10h后与清洗后的牛蒡、莲藕和清洗去核后的红枣混合,按照原料水=1:10-15的比例混合后磨浆,混合浆液于100-121℃条件下灭菌5-20min后待用。
6.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(6)中接种曲霉孢子数为1-5×106CFU/ml、搅拌速度60-120rpm、通氧量30-50L/h、发酵温度25-35℃、发酵时间30-90h。
7.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(7)中发酵液过板框过滤机过滤,滤液于110-121℃条件下灭菌1-5min后冷却至35℃待用;
在20-40%步骤(8)所述灭菌液中接种3-8%乳酸菌种子液,于30-38℃培养8-16h;在20-40%步骤(8)所述灭菌液中接种3-8%双歧杆菌种子液,于30-38℃培养8-16h;在剩余20-60%步骤(8)所述灭菌液中接种3-8%酵母菌种子液,于22-33℃培养15-25h;发酵结束后将三者混合均匀;
所述步骤(9)中混合发酵液与一定比例的白砂糖和/或蜂蜜和/或低聚糖进行调配,其重量比为白砂糖:蜂蜜:低聚果糖=100:10-30:10-30。
8.根据权利要求1所述的一种富含黄酮苷元和活性益生菌牛蒡酵素生产方法,其特征在于所述步骤(10)中牛蒡酵素的稳态化处理条件为:将海藻糖、羧甲基纤维素钠和黄原胶溶解于降温至30℃左右的凉开水中,然后与二次混合发酵液充分混合,使海藻糖、羧甲基纤维素钠和黄原胶在终产品中浓度达到1-3%、0.2-1%和0.05-0.5%;利用真空充氮包装机进行抽真空和充氮,使气体置换率达到98%以上后进行包装。
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