CN108433043A - 一种具有特殊风味的复合乳酸菌发酵肉脯及其制备方法 - Google Patents
一种具有特殊风味的复合乳酸菌发酵肉脯及其制备方法 Download PDFInfo
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- CN108433043A CN108433043A CN201810062498.2A CN201810062498A CN108433043A CN 108433043 A CN108433043 A CN 108433043A CN 201810062498 A CN201810062498 A CN 201810062498A CN 108433043 A CN108433043 A CN 108433043A
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Classifications
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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Abstract
本发明公开了一种具有特殊风味的复合乳酸菌发酵肉脯及其制备方法，其以鸭肉与鱼肉复合肉糜为主要原料，添加乳酸菌发酵剂，经混合、腌制、发酵、成熟、冷却、真空包装制作而成，所述乳酸菌发酵剂为嗜酸乳杆菌、戊糖片球菌混合而成；其制备方法包括以下步骤：先制备肉糜，再制备发酵剂，配置肉脯馅料，控制发酵过程，再将肉脯成熟干燥，真空包装，即可。本发酵肉脯具有独特的风味和口感，发酵剂的添加促进了脂肪和蛋白质的分解，增加了产品的风味，消除了鸭肉及鱼肉的腥味，且通过发酵改善了肉脯的组织结构，抑制了有害微生物的生长及脂肪的氧化，降低了亚硝酸盐的残留量，显著提高了产品的贮藏期及安全性。
Description
技术领域
本发明属于肉脯加工技术领域，具体涉及一种具有特殊风味的复合乳酸菌发酵肉脯及其制备方法。
背景技术
鸭肉具有低脂肪、低胆固醇、高蛋白等优点，深受广大消费者喜爱，其可食部分蛋白质含量高达20％～25％，同时，鸭肉风味独特，易于消化吸收，所含B 族维生素和维生素E较其他肉类多，能有效抵抗脚气病，神经炎和多种炎症，还有抗氧化抗衰老的作用。
鱼肉含有丰富的完全蛋白质、不饱和脂肪酸、含有丰富的磺，还含有磷、钙、铁等无机盐。鱼肉还含有大量的维生素A、维生素D、维生素B1、尼克酸等人体需要的营养素。
肉脯是我国三大干肉制品之一，根据其原料的处理方式可以分为肉糜脯和肉脯两类，顾名思义，肉糜脯是原料肉经绞碎、拌料、腌制等一系列工序制作而成；肉脯则是以去除筋腱的猪、牛等瘦肉为原料，经切片、调味、腌制等多道工序制作而成。肉脯中不但含有丰富的蛋白质、脂溶性维生素等营养成分，而且具有其独特的风味和口感，同时也具有食用方便和便于携带等优点。
乳酸菌为***，是最早从发酵肉制品中分离得到的菌属，由于其具有较强的耐酸性、能够抑制致病菌和腐败菌生长繁殖，部分乳酸菌还可以分泌蛋白酶和脂肪酶促进蛋白质和脂肪的分解，产生肽、氨基酸等小分子化合物，改善产品的风味，同时，乳酸菌作为肉制品发酵剂在一定程度上还可以改变肉制品的组织结构，使其口感更佳松软。
发明内容
针对现有问题的不足，本发明的目的是提供一种具有特殊风味的复合乳酸菌发酵肉脯及其制备方法；该发酵肉脯具有独特的风味和口感，发酵剂的添加促进了脂肪和蛋白质的分解，增加了产品的风味，消除了鸭肉及鱼肉的腥味，且通过发酵改善了肉脯的组织结构，抑制了有害微生物的生长及脂肪的氧化，降低了亚硝酸盐的残留量，显著提高了产品的贮藏期及安全性。
本发明解决其技术问题采用的技术方案是：
一种具有特殊风味的复合乳酸菌发酵肉脯，以鸭肉与鱼肉复合肉糜为主要原料，添加乳酸菌发酵剂，经混合、腌制、发酵、成熟、冷却、真空包装制作而成，所述乳酸菌发酵剂为嗜酸乳杆菌、戊糖片球菌混合而成。选用嗜酸乳杆菌、戊糖片球菌主要目的是产生良好的风味，消除腥味。
作为本申请的优选技术方案，其组分及对应的质量分数为：鸭鱼复合肉糜 100％、乳酸菌发酵剂106cfu/g-107cfu/g、食盐1.5-2.0％、酱油1.0-1.5％、红曲粉 0.10-0.30％、谷胺酰胺转氨酶0.2-0.4％、红曲粉0.10-0.30％、大豆异黄酮 0.01-0.03％、亚硝酸盐0.01-0.03％、白糖3.0-5.0％、料酒2.0-2.5％、五香粉0.5-1.0％、生姜粉0.3-0.5％、白胡椒粉0.5-1.0％、辣椒粉0.20-0.35％。
一种具有特殊风味的复合乳酸菌发酵肉脯的制备方法，包括以下步骤：
(1)鸭肉糜的制备：选择符合国家卫生标准的鸭胸肉，并去除筋腱和可见脂肪，将鸭肉清洗，切块，绞肉机绞碎，制成鸭肉糜；
(2)鱼肉糜的制备：将新鲜的鱼宰杀后，除去鱼头、鱼尾及鱼鳞及鱼皮，清洗干净，去除鱼骨、刺及鱼皮，用绞肉机将鱼肉绞碎，制得鱼肉糜；
(3)乳酸菌发酵剂的制备：先将菌株活化，再进行培养，制备发酵剂；
(4)肉脯馅料的混合及腌制：将鸭肉与鱼肉按(60-80％)：(20-40％)比例复配制成混合肉糜，以混合肉糜质量为基准，添加辅料，并加入制备好的复合乳酸菌发酵剂，混合均匀，于4-8℃温度下腌制2-3h后，置于模具内抹平至2-3mm厚度；将鸭肉和鱼肉复配支配混合肉糜，营养成分互为补充，鸭肉蛋白与鱼肉蛋白相互作用改善肉脯的质构；
(5)发酵过程的控制：调整发酵箱温度为20-24℃、将抹平后的肉脯配料放入发酵箱内进行发酵24-48h；
(6)发酵肉脯的成熟：先将发酵后的肉脯混合配料放于鼓风干燥箱中脱水，然后放置于温度为160-180℃的烤箱中烤制3-6min；
(7)真空包装：待肉脯冷却后，将肉脯切成小块，真空包装即可。
作为本申请的优选技术方案，所述步骤(3)包括如下具体步骤：
a.菌株的活化
将-80℃保存的戊糖片球菌和嗜酸乳杆菌进行复苏，划线培养于MRS固体培养基中，并于37℃恒温培养箱中培养18-24h，挑取菌落转接于小试中培养至一定浓度后，按照1.0-3.0％接种量接种于含有MRS液体培养液的锥形瓶中；
b.发酵剂的制作
根据戊糖片球菌和嗜酸乳杆菌的发酵生长特性，分别于37℃培养12-24h至活菌数达到106cfu/mL-107cfu/mL，并取嗜酸乳杆菌发酵剂：戊糖片球菌发酵剂＝1-2:1 混合复配，离心分离，用0.9％无菌生理盐水悬浮备用，得到复合乳酸菌发酵剂。
优选的，所述步骤(b)中，离心分离的转速为4000-6000r/min，时间10-15min。
步骤(3)中MRS液体培养基，配方和制备方法如下：蛋白胨l0g，牛肉膏 l0g，酵母提取物5g，K2HPO4 2g，柠檬酸二铵2g，乙酸钠5g，葡萄糖20g，吐温-80 1mL，MgSO4·7H2O 0.5g，MnSO40.25g，蒸馏水1000mL，pH6.2～6.4，121℃灭菌20min；MRS固体培养基，则在液体培养基配方的基础上加入1.5％的琼脂。
有益效果
本发明首次公开了一种具有特殊风味的复合乳酸菌发酵发酵肉脯及其制备方法，在肉脯的制作中加入红曲粉及通过乳酸菌发酵显著降低了亚硝酸盐的使用量和残留量，而且乳酸菌发酵产生的产物及添加的红曲粉具有抗氧化等保健功能，提高了肉脯的品质；肉脯中TG酶的添加及鱼肉与鸭肉的复合，改善了产品的质构，其必需氨基酸及不饱和脂肪酸等营养组分高于普通畜禽肉制作的肉脯。
另外，乳酸菌发酵过程中分泌的蛋白酶和脂肪酶促进脂肪及蛋白质等的分解，产生风味物质，同乳酸菌的发酵能消除鸭肉和鱼肉的腥味，从而改善了肉脯的风味和口感，并使肉脯组织结构更为疏松柔软；乳酸菌分泌的Beta-葡糖苷酶能把大豆异黄酮中溶解性比较低的黄酮苷元转化为溶解性比较高的黄酮苷，从而提高了其生物活性；同时乳酸菌发酵过程中产生的乳酸降低产品的pH值，抑制了有害微生物的生长；发酵过程中产生的抗氧化物质，抑制了脂肪的氧化，从而延长了产品的贮藏期。
附图说明
图1为未添加乳酸菌发酵剂的肉脯1000倍下组织结构示意图；
图2为添加乳酸菌发酵剂的肉脯1000倍下组织结构示意图。
具体实施方式
以下结合实施例对本发明做进一步详细说明。所用试剂或者仪器设备未注明生产厂商的，均视为可以通过市场购买的常规产品。
实施例1
依照以下步骤制作乳酸菌发酵肉脯：
1.鸭肉糜的制备：选择符合国家卫生标准的鸭胸肉，并去除筋腱和可见脂肪，将鸭肉用水进行清洗，切块，绞肉机绞碎，制成鸭肉糜；
2.鱼肉的制备：将新鲜的鱼宰杀后，出去鱼头、鱼尾及鱼鳞及鱼皮，清洗干净，去除鱼骨、刺及鱼皮，用绞肉机鱼肉绞碎，制得鱼肉糜；
3.乳酸菌发酵剂的制备，主要包括菌株的活化、发酵剂的制作；
(1)菌株的活化
将-80℃保存的戊糖片球菌和嗜酸乳杆菌进行复苏，划线培养于MRS固体培养基中，并于37℃恒温培养箱中培养22-24h，挑取菌落转接于小试中培养至一定浓度后，按照1.0-1.50％接种量接种于含有MRS液体培养液的锥形瓶中。
(2)发酵剂的制作
根据戊糖片球菌和嗜酸乳杆菌的发酵生长特性，分别于37℃培养12-18h至活菌数达到106cfu/mL，并取嗜酸乳杆菌发酵剂：戊糖片球菌发酵剂＝1:1混合复配，4000-4500r/min离心15min，用0.9％无菌生理盐水悬浮备用，得到复合乳酸菌发酵剂。
4.肉脯馅料的配置：将鸭肉与鱼肉按60％：40％比例复配制成混合肉糜，以混合肉糜为基准，添加食盐1.5-2.0％、白糖3.0-5.0％、料酒2.0-2.5％、酱油 1.0-1.5％、五香粉0.5-1.0％、生姜粉0.3-0.5％、白胡椒粉0.5-1.0％、辣椒粉 0.20-0.35％、谷胺酰胺转氨酶(简称TG酶)0.2-0.4％、红曲粉0.10-0.30％、亚硝酸盐0.01-0.03％、大豆异黄酮0.01-0.03％，并按照106cfu/g的浓度加入制备好的复合乳酸菌发酵剂，混合均匀，于4-5℃温度下腌制3h后，置于模具内抹平至 3mm厚度；
5.发酵过程的控制：调整发酵箱温度为20℃、将抹平后的肉脯脯配料放入发酵箱内进行发酵40-48h；
6.发酵鸭肉脯的成熟：先将发酵后的鸭肉脯放于60℃鼓风干燥箱中进行脱水2h，然后放置于温度为160℃的烤箱中烤制6min；
7.真空包装：待鸭肉脯冷却后，将肉脯切成50mm*35mm的小块，进行真空包装即可。
实施例2
除步骤3(1)中于37℃恒温培养箱中培养18-20h并按照2.0-3.0％接种量接种于含有MRS液体培养液的锥形瓶中外，其他同实施例1。
实施例3
除步骤3(2)中分别于37℃培养20-24h至活菌数达到107cfu/mL，并取嗜酸乳杆菌发酵剂：戊糖片球菌发酵剂＝2:1混合复配，5000-6000r/min离心10min 外，其他同实施例1。
实施例4
除步骤4将鸭肉与鱼肉按70％：30％比例复配制成混合肉糜，并按照107cfu/g 的浓度加入制备好的复合乳酸菌发酵剂外，其他同实施例1。
实施例5
除步骤4将鸭肉与鱼肉按80％：20％比例复配制成混合肉糜外，其他同实施例1。
将上述实施例制得的肉脯进行理化性能评价，具体为：
1.戊糖片球菌、嗜酸乳杆菌脂肪酶和蛋白酶分泌能力的测定
(1)蛋白酶活性的测定：MRS固体培养基中添加15％脱脂乳粉作为蛋白酶的检测培养基，无菌条件下分别吸取0.1mL嗜酸乳杆菌和戊糖片球菌，且分别均匀涂布于蛋白酶检测培养基中，37℃培养48h后观察实验结果，若菌落周围有透明环状物质生成，则为阳性，证明该菌株具有蛋白酶活性，反之亦然。
(2)脂肪酶活性的测定：MRS固体培养基中添加15％的猪油和中性红指示剂作为脂肪酶的检测培养基，在无菌条件下分别吸取0.1mL嗜酸乳杆菌和戊糖片球菌，且分别均匀涂布于脂肪酶检测培养基中，37℃培养48h后观察实验结果，若培养基上出现红色斑点，则为阳性，说明该菌株具有脂肪酶活性，反之亦然。
其试验结果如下表所示：
注：+表示阳性，-表示阴性
2.肉脯组织结构的测定
(1)肉脯样品的处理：取4℃保存的肉脯和发酵肉脯样品，经过3％戊二醛前固定，后用0.1M磷酸盐缓冲液漂洗，1％锇酸后固定，再次用0.1M磷酸盐缓冲液漂洗，再经乙醇梯度脱水(乙醇浓度设置为30％、50％、70％、90％、100％)， -40℃预冻48h，再经-80℃冷冻干燥24h后备用。
(2)取冷冻干燥的肉脯样品，剪成5mm*5mm小块，经离子溅射仪镀膜后放于扫描电子显微镜下观察实验结果。
其试验结果如图1、图2所示：
其中图1为未添加乳酸菌发酵剂的肉脯1000倍下组织结构示意图，图2为添加乳酸菌发酵剂的肉脯1000倍下组织结构示意图，从图中可以看出：经过乳酸菌发酵的肉脯其组织结构松软，结构破坏较为明显。
3.亚硝酸盐含量的测定
亚硝酸含量的测定参照萘乙二胺分光光度法进行，其测定步骤如下：
(1)样品预处理和提取：取50g肉脯样并切成小块，置于匀浆机中，加入蒸馏水50mL并制成匀浆，称取5g(精确至0.01g)制成匀浆的试样，置于50mL 烧杯中，加12.5mL饱和硼酸溶液搅拌均匀，以70℃左右的水约300mL将试样洗入500mL容量瓶中，沸水浴中加热15min，取出置冷水浴中冷却，并放置至室温。
(2)提取液的净化：振荡提取液时，加入5mL亚铁***摇匀后再加5mL 乙酸锌，以沉淀蛋白质，加水至刻度，摇匀，静置30min，除去上层脂肪，清液用滤纸过滤，弃去初滤液30mL，滤液备用，对照组为未添加发酵剂的肉脯，其处理方法同上。
(3)亚硝酸盐的测定：于25mL带塞比色管中加入2mL NaNO2标准使用液，反应10min、分别加入一定量(2)中两种滤液，立即加入1mL0.4％对氨基苯磺酸混合均匀，静置3-5min，加入0.5mL 0.2％盐酸萘乙二胺，振荡混合，再加蒸馏水定容至25m L，静置15min，以相应的试样空白调零，测定538nm波长处吸光度，亚硝酸盐含量的清除率按式计算：
清除率/％＝(A对照－A发酵)/A对照×100
式中：A对照为不添加发酵剂的肉脯样品，A发酵为添加发酵剂的肉脯样品，结果显示亚硝酸盐的清除率为59.7％。
4.肉脯在贮藏过程中菌落总数的测定
乳酸菌菌落总数的测定主要包括菌落总数、霉菌和酵母菌的检验，其试验方法如下：
(1)菌落总数的检测方法：称取25g样品置于盛有225mL生理盐水的无菌均质杯内，10000r/min均质1min，制成1：10的样品匀液；无菌吸取1mlL的样品匀液，梯度稀释其浓度为1：10、1：102、1：103、1：104、1：105、选择2～3 个适宜稀释度的样品匀液，各取1mL分别放入平板计数琼脂培养基中进行培养，其培养温度为36±1℃，培养时间为48h，每个梯度两个平行；然后进行计算各平板菌落总数，选取菌落总数在30～300cfu之间、无蔓延菌落生长的平板计数菌落总数。
(2)霉菌和酵母菌的检测方法：称取25g样品置于盛有225mL生理盐水的无菌均质杯内，10000r/min均质1min，制成1：10的样品匀液；无菌吸取1mlL 的样品匀液，梯度稀释其浓度为1：10、1：102、1：103、1：104、1：105、选择 2～3个适宜稀释度的样品匀液，各取1mL分别放入马铃薯葡萄糖琼脂培养基中进行培养，其培养温度为28±1℃，培养时间为5d，每个梯度两个平行；然后根据菌落形态分别计数霉菌和酵母，选取菌落数在10～150cfu之间、无蔓延菌落生长的平板计算霉菌和酵母菌的数量。
通过测定发酵肉脯和未添加乳酸菌发酵剂的肉脯其菌落总数、霉菌和酵母菌落数，其试验结果表明，在同等贮藏条件下，发酵组细菌总数在贮藏30天后，其菌落总数为1.9*104cfu/g，霉菌和酵母菌总数为2.4*103cfu/g，而未添加发酵剂的肉脯其菌落总数为7.8*104cfu/g，霉菌和酵母菌总数为1.3*104cfu/g，由此可说明发酵剂的添加提高了肉脯的贮藏特性，保证了产品的安全卫生。
本发明的保护内容不局限于以上实施例。在不背离发明构思的精神和范围下，本领域技术人员能够想到的变化和优点都被包括在本发明中，并且以所附的权利要求为保护范围。
Claims (5)
1.一种具有特殊风味的复合乳酸菌发酵肉脯，其特征在于，以鸭肉与鱼肉复合肉糜为主要原料，添加乳酸菌发酵剂，经混合、腌制、发酵、成熟、冷却、真空包装制作而成，所述乳酸菌发酵剂为嗜酸乳杆菌、戊糖片球菌混合而成。
2.根据权利要求1所述的具有特殊风味的复合乳酸菌发酵肉脯，其特征在于，其组分及对应的质量分数为：鸭鱼复合肉糜100％、乳酸菌发酵剂106cfu/g-107cfu/g、食盐1.5-2.0％、酱油1.0-1.5％、红曲粉0.10-0.30％、谷胺酰胺转氨酶0.2-0.4％、红曲粉0.10-0.30％、大豆异黄酮0.01-0.03％、亚硝酸盐0.01-0.03％、白糖3.0-5.0％、料酒2.0-2.5％、五香粉0.5-1.0％、生姜粉0.3-0.5％、白胡椒粉0.5-1.0％、辣椒粉0.20-0.35％。
3.权利要求1所述的具有特殊风味的复合乳酸菌发酵肉脯的制备方法，其特征在于，包括以下步骤：
(1)鸭肉糜的制备：选择符合国家卫生标准的鸭胸肉，并去除筋腱和可见脂肪，将鸭肉清洗，切块，绞肉机绞碎，制成鸭肉糜；
(2)鱼肉糜的制备：将新鲜的鱼宰杀后，除去鱼头、鱼尾及鱼鳞及鱼皮，清洗干净，去除鱼骨、刺及鱼皮，用绞肉机将鱼肉绞碎，制得鱼肉糜；
(3)乳酸菌发酵剂的制备：先将菌株活化，再进行培养，制备发酵剂；
(4)肉脯馅料的混合及腌制：将鸭肉与鱼肉按(60-80％)：(20-40％)比例复配制成混合肉糜，以混合肉糜质量为基准，添加辅料，并加入制备好的复合乳酸菌发酵剂，混合均匀，于4-8℃温度下腌制2-3h后，置于模具内抹平至2-3mm厚度；
(5)发酵过程的控制：调整发酵箱温度为20-24℃、将抹平后的肉脯配料放入发酵箱内进行发酵24-48h；
(6)发酵肉脯的成熟：先将发酵后的肉脯混合配料放于鼓风干燥箱中脱水，然后放置于温度为160-180℃的烤箱中烤制3-6min；
(7)真空包装：待肉脯冷却后，将肉脯切成小块，真空包装即可。
4.根据权利要求3所述的具有特殊风味的复合乳酸菌发酵肉脯的制备方法，其特征在于，所述步骤(3)包括如下具体步骤：
a.菌株的活化
将-80℃保存的戊糖片球菌和嗜酸乳杆菌进行复苏，划线培养于MRS固体培养基中，并于37℃恒温培养箱中培养18-24h，挑取菌落转接于小试中培养至一定浓度后，按照1.0-3.0％接种量接种于含有MRS液体培养液的锥形瓶中；
b.发酵剂的制作
根据戊糖片球菌和嗜酸乳杆菌的发酵生长特性，分别于37℃培养12-24h至活菌数达到106cfu/mL-107cfu/mL，并取嗜酸乳杆菌发酵剂：戊糖片球菌发酵剂＝1-2:1混合复配，离心分离，用0.9％无菌生理盐水悬浮备用，得到复合乳酸菌发酵剂。
5.根据权利要求4所述的具有特殊风味的复合乳酸菌发酵肉脯的制备方法，其特征在于，所述步骤(b)中，离心分离的转速为4000-6000r/min，时间10-15min。
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Application publication date: 20180824 |