CN109007758A - 一种富硒螺旋藻酱油的加工方法 - Google Patents
一种富硒螺旋藻酱油的加工方法 Download PDFInfo
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- CN109007758A CN109007758A CN201810903537.7A CN201810903537A CN109007758A CN 109007758 A CN109007758 A CN 109007758A CN 201810903537 A CN201810903537 A CN 201810903537A CN 109007758 A CN109007758 A CN 109007758A
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- selenium
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- soy sauce
- aspergillus oryzae
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
- A23L27/105—Natural spices, flavouring agents or condiments; Extracts thereof obtained from liliaceae, e.g. onions, garlic
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/21—Removal of unwanted matter, e.g. deodorisation or detoxification by heating without chemical treatment, e.g. steam treatment, cooking
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
本发明公开了一种富硒螺旋藻酱油的加工方法,其特征在于方法步骤如下:步骤1、米曲霉菌种驯化:具体为米曲霉的工艺驯化;①菌种活化;②紫外诱变;③初筛;④复筛;⑤稳定性检测;步骤2溶菌酶处理富硒螺旋藻藻浆;步骤3制曲;步骤4无菌制醪;步骤5板框压滤提油;步骤6酱油抗氧化调味类制备;步骤7酱油调味;步骤8富硒白酱油灌装。本发明优点:本发明生产富硒螺旋藻白酱油,采用天然抗氧化活性成分抑制美拉德反应对氨基酸效价的破坏;产品除了具有含丰富有机硒、产品中有效氨基酸含量高、低盐、无菌、食品安全性好的优势之外,还具有淡淡蒜香和浓厚酱香的营养调味料。
Description
技术领域
本发明属于农产品加工技术领域,涉及一种富硒螺旋藻酱油的加工方法。
背景技术
酱油是以大豆、小麦等原料,经过原料预处理、制曲、发酵、浸出淋油及加热配制等工艺生产出来的调味品,营养极其丰富,主要营养成份包括氨基酸、可溶性蛋白质、糖类、有机酸类等,色泽红褐色,有独特酱香,滋味鲜美,有助于促进食欲。近些年来, 我国的酱油行业迅猛发展,出现了大量以水加味精及合成色素和防腐剂勾兑而成的配制酱油,长期食用对肝、肾有一定毒副作用,甚至还会影响儿童智力发育。另外,传统的酱油生产过程中,会有菌种混杂,甚至杂菌带来食品安全风险,酿制过程中的依赖高盐分和化学合成的防腐剂保鲜。传统酱油过渡追求色泽,以致需要长达1年的室外日晒以求美拉德反应深度进行,蚊蝇滋生,其结果是导致酱油中氨基酸效价严重下降、卫生状况堪忧,需要在接受现代食品科学的研究成果,逐步改变观念。
硒(Se)是人体必须的微量元素,在人体中发挥着重要的生理功能,被国内外医药界和营养学界尊称为“生命的火种”,享有“长寿元素”、“防癌之王”、“心脏守护神”、“天然解毒剂”等美誉。其主要原理是:第一、组成体内抗氧化酶,能保护细胞膜免受氧化损伤,保持其通透性;第二、硒-P蛋白具有螯合重金属等毒物,降低毒物毒性作用。硒元素的缺乏将导致多种疾病的发生。因此世界卫生组织建议每天补充200 μg硒,可有效预防多种疾病的高发。“硒”是人体必需的,又不能自制,又因为无机硒盐毒性较大,因此仅靠天然食物的摄入无法满足人类的正常需要。营养学家提倡通过硒营养强化食物补充有机硒。螺旋藻富含蛋白,还含有多种对人体有益的物质如脂肪酸、活性肽、维生素等,具有抗氧化、防肿瘤,降低胆固醇,调节血糖和增强免疫系统的功效。目前,已有利用螺旋藻生产酱油,但是由于螺旋藻含有特殊的藻腥味,系螺旋藻干燥方法不当引发会影响食品的口感和味道,没有被市场广泛接受。另外,现有的螺旋藻酱油中不含有硒元素,对螺旋藻的利用也不够充分。人们对食品的要求随着生活质量的提高而升高,不仅仅局限在色泽和味道的好坏,更多的是追求其营养和保健功能。
发明内容
本发明的目的在于提出一种富硒螺旋藻酱油的加工方法,该加工方法优选富硒地带生产的富硒大豆豆粕、富硒小麦和采用富硒生产工艺生产的螺旋藻作为生产原料,采用现代食品加工工艺制造而成富硒螺旋藻酱油。
本发明采用的技术方案如下:一种富硒螺旋藻酱油的加工方法,其特征在于方法步骤如下:
步骤1、米曲霉菌种驯化:具体为米曲霉的工艺驯化;
①菌种活化;从国家菌种保藏库中购买编号为BNCC338270的米曲霉进行试管斜面活化,于37oC恒温培养3d,获得生长正常、孢子丰富的斜面培养物;
②紫外诱变;取1mL单袍子悬液于直径6cm的无菌培养皿中,调整液体厚度不超过1mm,距30W紫外灯15cm处照射15min,同时晃动培养皿;
③初筛;传代三次,用活化的米曲霉菌株制孢子悬浮液;取米曲霉孢子悬液0.1mL涂布在硒浓度为140 μg/mL的察氏平板培养基上,37oC避光培养72h,选择周围有红色硒单质出现的菌落,将其分别接种到查氏斜面培养基保存;
④复筛;取初筛保存的米曲霉斜面一支,从中挑取一环抱子,接种到种子培养基中,置于180 r/min的揺床上,37oC,培养48h;然后配制造浓度为40、50、60、70、80、90、100 μg/mL的YEPD培养基,按8%的接种量接种初筛的菌种,37oC培养48h,通过测量各组生物量和蛋白酶活性值,选出最强的米曲霉菌株;
⑤稳定性检测;将复筛得到的菌株传代培养3次,测量每次传代培养菌种的生物量和蛋白酶活性,确定优先的菌株,以优选的菌株依次扩大为母种、再扩大培养制备米曲霉生产种;
步骤2、溶菌酶处理富硒螺旋藻藻浆:每100份的新鲜螺旋藻藻浆经过常压煮沸冷却到室温,添加5份的鸡蛋蛋清蛋白混合均匀,在37℃下酶解8~12h;
步骤3、制曲:按照富硒大豆豆粕:富硒小麦:溶菌酶预处理后的富硒螺旋藻浆为5:3:2的原料配比,润水后蒸料30min,取出后冷却,按接种量0.3%接种经过驯化的米曲霉,曲盘中铺平,放在30℃的恒温恒湿无菌培养室的培养基架上培养制曲48h,期间进行搅拌防止干曲;
步骤4、无菌制醪:采用在无菌陶缸内操作,拌曲盐水浓度为4~6 °Bé,经过煮沸杀菌冷却到室温、拌曲盐水量约为成曲量的2.5倍,要求稀盐水与成曲搅拌均匀;随后逐渐使培养室温度上升到45℃,维持6~12小时,促进曲霉细胞自溶释放蛋白酶、淀粉酶以及纤维素酶;再将室温逐渐下调到35℃,继续酶解1~2周,检测固体残渣中粗蛋白含量小于1%时,即表明酱醪成熟;
步骤5、板框压滤提油:成熟酱醪采用板框压滤机压滤提取酱油,选用200目以上滤布确保酱油澄清无固体残渣混入,得到富硒白酱油原浆;
步骤6、酱油抗氧化调味类制备:富硒大蒜蒜头30份,除去外皮和根部残余物,以打浆机粉碎,空气中静置3~5小时,促进大蒜精油形成;生姜10份洗净,以打浆机粉碎,有机绿茶5份,有机鲜荷叶5份切碎;4种成分混合,添加50份水置于高压锅中煮熟冷却,把汁滤出备用;
步骤7、酱油调味:每99份步骤5富硒白酱油原浆,添加1份步骤6调味液,混合均匀,经澄清后制得富硒白酱油初品;
步骤8、富硒白酱油灌装:富硒白酱油初品采用低温等离子体灌装设备,250ml玻璃无菌灌装,得到富硒白酱油成品,供市场销售,保质期1年。
本发明所述步骤1中斜面培养基(察氏培养基)配方为:硝酸钠3g,磷酸二氢钾1g,硫酸镁0.5g,氯化钾0.5g,硫酸亚铁0.01g,蔗糖30g,琼脂20g,pH自然,加水定容到1L,121oC灭菌30min。
本发明所述步骤1中平板培养基配方为:在斜面培养基中添加亚硒酸钠0.14g,加水定容至1L,121oC灭菌30min。
本发明所述步骤1中种子培养基配方为:豆芽30g煮成汁,可溶性淀粉20g,磷酸二氢钾1g,硫酸胺0.5g,硫酸镁0.5g,亚硒酸钠0.1g,pH值自然,加水定容到1L,121oC灭菌30min。
本发明所述步骤1中发酵培养基(YEPD)配方为:葡萄糖20g,酵母膏20g,硫酸按5g,磷酸二氢钾1g,氯化钙1g,硫酸镁0.5g,维生素B10.015g,亚硒酸0.1g,加水定容到1L,121oC灭菌30min。
本发明从原料选择、菌种优选、工艺现代化说明:
1、原料选择:优选富硒地带生产的富硒大豆豆粕、富硒小麦和采用富硒生产工艺生产的螺旋藻作为生产原料。对富硒螺旋藻藻浆免除热风干燥、直接以溶菌酶酶解后并入富硒大豆和小麦主料中蒸煮灭菌,消除螺旋藻腥味发生的根源。
2、菌种优选:优选蛋白酶、淀粉酶和纤维素酶功能强大的米曲霉进行定向驯化,在白酱油生产中实行纯种发酵制曲。
3、工艺现代化:酱醪成熟采用无菌工艺过程避免借助高盐分控制杂菌繁殖,优化熟化过程工艺参数、促进曲霉细胞内酶的释放和活性保持,把蛋白质水解时间缩短到最短;采用现代压滤工艺提高酱油得率,提升酱油氨基酸含量水平;采用低温等离子杀菌设备对白酱油实现无菌灌装,消除传统酱油高盐的缺点。
4、使用安全的富含多酚类、大蒜精油类抗氧化活性成分的调味料提取液对酱油进行调味,抑制美拉德反应对酱油品质产生不良影响。
本发明优点:本发明生产富硒螺旋藻白酱油,采用天然抗氧化活性成分抑制美拉德反应对氨基酸效价的破坏;产品除了具有含丰富有机硒、产品中有效氨基酸含量高、低盐、无菌、食品安全性好的优势之外,还具有淡淡蒜香和浓厚酱香的营养调味料。
具体实施方式
实施例1
以富硒大豆豆粕、富硒小麦、富硒螺旋藻为原料,在制曲过程中加入螺旋藻藻浆生产富硒螺旋藻酱油。
步骤1、米曲霉菌种驯化:(1)米曲霉的工艺驯化。①菌种活化。从国家菌种保藏库中购买编号为BNCC338270的米曲霉进行试管斜面活化,于37oC 恒温培养3d,获得生长正常、孢子丰富的斜面培养物。②紫外诱变。取1mL单袍子悬液于直径6cm的无菌培养皿中,调整液体厚度不超过1mm,距30W紫外灯15cm处照射15min,同时晃动培养皿。③初筛。传代三次,用活化的米曲霉菌株制孢子悬浮液。取米曲霉孢子悬液0.1mL涂布在硒浓度为140 μg/mL的察氏平板培养基上,37oC避光培养72h,选择周围有红色硒单质出现的菌落,将其分别接种到察氏斜面培养基保存。④复筛。取初筛保存的米曲霉斜面一支,从中挑取一环抱子,接种到种子培养基中,置于180 r/min的揺床上,37oC,培养48h。然后配制造浓度为40、50、60、70、80、90、100 μg/mL的YEPD培养基,按8%的接种量接种初筛的菌种,37oC培养48h,通过测量各组生物量和蛋白酶活性值,选出最强的米曲霉菌株。⑤稳定性检测。将复筛得到的菌株传代培养3次,测量每次传代培养菌种的生物量和蛋白酶活性,确定优先的菌株,以优选的菌株依次扩大为母种、再扩大培养制备米曲霉生产种。
(2)米曲霉生产种的扩大培养。①斜面培养。经过驯化的米曲霉优先菌种接种到斜面培养基上,于37oC 恒温培养3d。②种子培养。从保存的斜面培养基上取一环经紫外诱变过的米曲霉孢子,接种到250 mL的三角瓶,装液量为100 mL的种子培养基中,摇床转速180r/min,湿度37oC,培养48h。③扩大培养。培养基配方:
斜面培养基(察氏培养基):硝酸钠3g,磷酸二氢钾1g,硫酸镁0.5g,氯化钾0.5g,硫酸亚铁0.01g,蔗糖30g,琼脂20g,pH自然,加水定容到1L,121oC灭菌30min。
平板培养基:在斜面培养基中添加亚硒酸钠0.14g,加水定容至1L,121oC灭菌30min。
种子培养基配方:豆芽30g煮成汁,可溶性淀粉20g,磷酸二氢钾1g,硫酸胺0.5g,硫酸镁0.5g,亚硒酸钠0.1g,pH值自然,加水定容到1L,121oC灭菌30min。
发酵培养基(YEPD):葡萄糖20g,酵母膏20g,硫酸按5g,磷酸二氢钾1g,氯化钙1g,硫酸镁0.5g,维生素B10.015g,亚硒酸0.1g,加水定容到1L,121oC灭菌30min。
步骤2、溶菌酶处理富硒螺旋藻藻浆:每100份的新鲜螺旋藻藻浆经过常压煮沸冷却到室温,添加5份的鸡蛋蛋清蛋白混合均匀,在37℃下酶解8~12h;
步骤3、制曲:按照富硒大豆:富硒小麦:溶菌酶预处理后的富硒螺旋藻浆为5:3:2的原料配比,润水后蒸料30min,取出后冷却,按接种量0.3%接种经过驯化的米曲霉,曲盘中铺平,放在30℃的恒温恒湿无菌培养室的培养基架上培养制曲48h,期间进行搅拌防止干曲。
步骤4、无菌制醪:采用在无菌陶缸内操作,拌曲盐水浓度为4~6 °Bé,,经过煮沸杀菌冷却到室温、拌曲盐水量约为成曲量的2.5倍,要求稀盐水与成曲搅拌均匀。随后逐渐使培养室温度上升到45℃,维持6~12小时,促进曲霉细胞自溶释放蛋白酶、淀粉酶以及纤维素酶;再将室温逐渐下调到35℃,继续酶解1~2周,检测固体残渣中粗蛋白含量小于1%时,即表明酱醪成熟;
步骤5、板框压滤提油:成熟酱醪采用板框压滤机压滤提取酱油,选用200目以上滤布确保酱油澄清无固体残渣混入,得到富硒白酱油原浆;
步骤6、酱油抗氧化调味类制备:富硒大蒜蒜头30份,除去外皮和根部残余物,以打浆机粉碎,空气中静置3~5小时,促进大蒜精油形成;生姜10份洗净,以打浆机粉碎,有机绿茶5份,有机鲜荷叶5份切碎;4种成分混合,添加50份水置于高压锅中煮熟冷却,把汁滤出备用;
步骤7、酱油调味:每99份步骤5富硒白酱油原浆,添加1份步骤6调味液,混合均匀,经澄清后制得富硒白酱油初品。
步骤8、富硒白酱油灌装:富硒白酱油初品采用低温等离子体灌装设备,250ml玻璃无菌灌装,得到富硒白酱油成品,供市场销售,保质期1年。
实施例2
以富硒大豆豆粕、富硒小麦、富硒螺旋藻为原料,在制曲过程中加入螺旋藻藻浆生产富硒螺旋藻酱油。
步骤1、米曲霉菌种驯化:(1)米曲霉的工艺驯化。①菌种活化。从国家菌种保藏库中购买编号为BNCC338270的米曲霉进行试管斜面活化,于37oC 恒温培养3d,获得生长正常、孢子丰富的斜面培养物。②紫外诱变。取1mL单袍子悬液于直径6cm的无菌培养皿中,调整液体厚度不超过1mm,距30W紫外灯15cm处照射15min,同时晃动培养皿。③初筛。传代三次,用活化的米曲霉菌株制孢子悬浮液。取米曲霉孢子悬液0.1mL涂布在硒浓度为140 μg/mL的察氏平板培养基上,37oC避光培养72h,选择周围有红色硒单质出现的菌落,将其分别接种到察氏斜面培养基保存。④复筛。取初筛保存的米曲霉斜面一支,从中挑取一环抱子,接种到种子培养基中,置于180 r/min的揺床上,37oC,培养48h。然后配制造浓度为40、50、60、70、80、90、100 μg/mL的YEPD培养基,按8%的接种量接种初筛的菌种,37oC培养48h,通过测量各组生物量和蛋白酶活性值,选出最强的米曲霉菌株。⑤稳定性检测。将复筛得到的菌株传代培养3次,测量每次传代培养菌种的生物量和蛋白酶活性,确定优先的菌株,以优选的菌株依次扩大为母种、再扩大培养制备米曲霉生产种。
(2)米曲霉生产种的扩大培养。①斜面培养。经过驯化的米曲霉优先菌种接种到斜面培养基上,于37oC 恒温培养3d。②种子培养。从保存的斜面培养基上取一环经紫外诱变过的米曲霉孢子,接种到250 mL的三角瓶,装液量为100 mL的种子培养基中,摇床转速180r/min,湿度37oC,培养48h。③扩大培养。培养基配方:
斜面培养基(察氏培养基):硝酸钠3g,磷酸二氢钾1g,硫酸镁0.5g,氯化钾0.5g,硫酸亚铁0.01g,蔗糖30g,琼脂20g,pH自然,加水定容到1L,121oC灭菌30min。
平板培养基:在斜面培养基中添加亚硒酸钠0.14g,加水定容至1L,121oC灭菌30min。
种子培养基配方:豆芽30g煮成汁,可溶性淀粉20g,磷酸二氢钾1g,硫酸胺0.5g,硫酸镁0.5g,亚硒酸钠0.1g,pH值自然,加水定容到1L,121oC灭菌30min。
发酵培养基(YEPD):葡萄糖20g,酵母膏20g,硫酸按5g,磷酸二氢钾1g,氯化钙1g,硫酸镁0.5g,维生素B10.015g,亚硒酸0.1g,加水定容到1L,121oC灭菌30min。
步骤2、溶菌酶处理富硒螺旋藻藻浆:每100份的新鲜螺旋藻藻浆经过常压煮沸冷却到室温,添加5份的鸡蛋蛋清蛋白混合均匀,在37℃下酶解8~12h;
步骤3、制曲:按照富硒大豆低温豆粕:富硒小麦麸皮:溶菌酶预处理后的富硒螺旋藻浆为5:4:1的原料配比,润水后蒸料30min,取出后冷却,按接种量0.3%接种经过驯化的米曲霉,曲盘中铺平,放在30℃的恒温恒湿无菌培养室的培养基架上培养制曲48h,期间进行搅拌防止干曲。
步骤4、无菌制醪:采用在无菌陶缸内操作,拌曲盐水浓度为4~6 °Bé,,经过煮沸杀菌冷却到室温、拌曲盐水量约为成曲量的2.5倍,要求稀盐水与成曲搅拌均匀。随后逐渐使培养室温度上升到45℃,维持6~12小时,促进曲霉细胞自溶释放蛋白酶、淀粉酶以及纤维素酶;再将室温逐渐下调到35℃,继续酶解1~2周,检测固体残渣中粗蛋白含量小于1%时,即表明酱醪成熟;
步骤5、板框压滤提油:成熟酱醪采用板框压滤机压滤提取酱油,选用200目以上滤布确保酱油澄清无固体残渣混入,得到富硒白酱油原浆;
步骤6、酱油抗氧化调味类制备:富硒大蒜蒜头30份,除去外皮和根部残余物,以打浆机粉碎,空气中静置3~5小时,促进大蒜精油形成;生姜10份洗净,以打浆机粉碎,有机绿茶5份,有机鲜荷叶5份切碎;4种成分混合,添加50份水置于高压锅中煮熟冷却,把汁滤出备用;
步骤7、酱油调味:每99份步骤5富硒白酱油原浆,添加1份步骤6调味液,混合均匀,经澄清后制得富硒白酱油初品;
步骤8、富硒白酱油灌装:富硒白酱油初品采用低温等离子体灌装设备,250ml玻璃无菌灌装,得到富硒白酱油成品,供市场销售,保质期1年。
Claims (5)
1.一种富硒螺旋藻酱油的加工方法,其特征在于方法步骤如下:
步骤1、米曲霉菌种驯化:具体为米曲霉的工艺驯化;
①菌种活化;从国家菌种保藏库中购买编号为BNCC338270的米曲霉进行试管斜面活化,于37oC恒温培养3d,获得生长正常、孢子丰富的斜面培养物;
②紫外诱变;取1mL单袍子悬液于直径6cm的无菌培养皿中,调整液体厚度不超过1mm,距30W紫外灯15cm处照射15min,同时晃动培养皿;
③初筛;传代三次,用活化的米曲霉菌株制孢子悬浮液;取米曲霉孢子悬液0.1mL涂布在硒浓度为140 μg/mL的平板培养基上,37oC避光培养72h,选择周围有红色硒单质出现的菌落,将其分别接种到察氏斜面培养基保存;
④复筛;取初筛保存的米曲霉斜面一支,从中挑取一环抱子,接种到种子培养基中,置于180 r/min的揺床上,37℃,培养48h;然后配制造浓度为40、50、60、70、80、90、100 μg/mL的YEPD培养基,按8%的接种量接种初筛的菌种,37oC培养48h,通过测量各组生物量和蛋白酶活性值,选出最强的米曲霉菌株;
⑤稳定性检测;将复筛得到的菌株传代培养3次,测量每次传代培养菌种的生物量和蛋白酶活性,确定优先的菌株,以优选的菌株依次扩大为母种、再扩大培养制备米曲霉生产种;
步骤2、溶菌酶处理富硒螺旋藻藻浆:每100份的新鲜螺旋藻藻浆经过常压煮沸冷却到室温,添加5份的鸡蛋蛋清蛋白混合均匀,在37℃下酶解8~12h;
步骤3、制曲:按照富硒大豆豆粕:富硒小麦:溶菌酶预处理后的富硒螺旋藻浆为5:3:2的原料配比,润水后蒸料30min,取出后冷却,按接种量0.3%接种经过驯化的米曲霉,曲盘中铺平,放在30℃的恒温恒湿无菌培养室的培养基架上培养制曲48h,期间进行搅拌防止干曲;
步骤4、无菌制醪:采用在无菌陶缸内操作,拌曲盐水浓度为4~6 °Bé,经过煮沸杀菌冷却到室温、拌曲盐水量约为成曲量的2.5倍,要求稀盐水与成曲搅拌均匀;随后逐渐使培养室温度上升到45℃,维持6~12小时,促进曲霉细胞自溶释放蛋白酶、淀粉酶以及纤维素酶;再将室温逐渐下调到35℃,继续酶解1~2周,检测固体残渣中粗蛋白含量小于1%时,即表明酱醪成熟;
步骤5、板框压滤提油:成熟酱醪采用板框压滤机压滤提取酱油,选用200目以上滤布确保酱油澄清无固体残渣混入,得到富硒白酱油原浆;
步骤6、酱油抗氧化调味类制备:富硒大蒜蒜头30份,除去外皮和根部残余物,以打浆机粉碎,空气中静置3~5小时,促进大蒜精油形成;生姜10份洗净,以打浆机粉碎,有机绿茶5份,有机鲜荷叶5份切碎;4种成分混合,添加50份水置于高压锅中煮熟冷却,把汁滤出备用;
步骤7、酱油调味:每99份步骤5富硒白酱油原浆,添加1份步骤6调味液,混合均匀,经澄清后制得富硒白酱油初品;
步骤8、富硒白酱油灌装:富硒白酱油初品采用低温等离子体灌装设备,250ml玻璃无菌灌装,得到富硒白酱油成品,供市场销售,保质期1年。
2.根据权利要求1所述的一种富硒螺旋藻酱油的加工方法,其特征在于:所述步骤1中察氏斜面培养基配方为:硝酸钠3g,磷酸二氢钾1g,硫酸镁0.5g,氯化钾0.5g,硫酸亚铁0.01g,蔗糖30g,琼脂20g,pH自然,加水定容到1L,121oC灭菌30min。
3.根据权利要求1所述的一种富硒螺旋藻酱油的加工方法,其特征在于:所述步骤1中平板培养基配方为:在斜面培养基中添加亚硒酸钠0.14g,加水定容至1L,121oC灭菌30min。
4.根据权利要求1所述的一种富硒螺旋藻酱油的加工方法,其特征在于:所述步骤1中种子培养基配方为:豆芽30g煮成汁,可溶性淀粉20g,磷酸二氢钾1g,硫酸胺0.5g,硫酸镁0.5g,亚硒酸钠0.1g,pH值自然,加水定容到1L,121oC灭菌30min。
5.根据权利要求1所述的一种富硒螺旋藻酱油的加工方法,其特征在于:所述步骤1中YEPD培养基配方为:葡萄糖20g,酵母膏20g,硫酸按5g,磷酸二氢钾1g,氯化钙1g,硫酸镁0.5g,维生素B10.015g,亚硒酸0.1g,加水定容到1L,121oC灭菌30min。
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