CN111820349A - 一种直投式果蔬汁乳酸菌促进剂 - Google Patents
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Abstract
本发明公开了一种直投式果蔬汁乳酸菌促进剂。其技术方案是：以保加利亚乳杆菌、嗜酸乳杆菌或嗜热链球菌作为发酵菌株，接种至增殖培养基中，控流分批补料培养，乳酸菌菌体经无菌蒸馏水离心洗涤两次后，与溶于无菌蒸馏水的冻干保护剂相混合制成菌悬液，经低温预冻及真空冷冻干燥处理，可得直投式果蔬汁乳酸菌促进剂。该促进剂具有活菌含量高、保质期长、便于存储和运输的优点。
Description
技术领域
本发明公开了一种直投式果蔬汁乳酸菌促进剂，属于生物制备领域。
背景技术
我国是世界最大的果蔬原料生产国，然而目前对果蔬的研究处于高产量、低加工水平，这导致我国新鲜水果腐败率达到30%，蔬菜腐败率达到40-50%，果蔬损耗率远远高于发达国家，因此对果蔬的深加工研究至关重要。利用乳酸菌促进剂发酵果蔬汁能够使饮料既有果蔬本身的香气，又可避免发酵饮品因巴氏杀菌等工艺而造成营养成分的损失，同时发酵促进剂能够改善果蔬汁的风味，使果蔬营养与乳酸菌的功能高效地融为一体。
目前，发酵饮料中常用的发酵剂需经过活化、扩培等系列过程，生产周期较长，且在加工过程中容易造成菌种污染、失活等弊端，不易于保存和运输，生产成本较高。
本发明将乳酸菌菌种接种至增殖培养基中，将乳酸菌菌体与冻干保护剂混合，经低温预冻及真空冷冻干燥处理可得到一种用于果蔬汁发酵的直投式促进剂。结合乳酸菌生长特点，采取前期低速流、后期高速流向培养基中添加植物成分，以改善底物营养，同时，植物成分对大部分植物乳杆菌的增菌效果优于其它，且植物成分的利用能够增加植物资源利用率。冻干保护剂的添加能够减少乳酸菌在冻干过程中的损失，且冻干前对乳酸菌进行胁迫处理能够引起细胞膜的应激变化，利于乳酸菌冻干存活率的提高。该乳酸菌促进剂能够直接接种至发酵基料中生产发酵果蔬汁，可有效减少污染发生几率，简化生产工艺。本促进剂活菌含量高，便于存储和运输，适合工业化生产。
发明内容
本发明公开了一种直投式果蔬汁乳酸菌促进剂。该促进剂可直接用于果蔬汁的发酵，活菌含量较高、便于存储和运输，适合工业化生产。
本发明的目的是通过以下技术方法实现的：
A、发酵菌株接种至增殖培养基中控流分批补料培养；
B、利用无菌蒸馏水离心洗涤乳酸菌菌体两次；
C、冻干保护剂溶于无菌蒸馏水后，与乳酸菌菌体混合制备菌悬液；
D、采用真空冷冻干燥制得直投式果蔬汁乳酸菌促进剂。
A中发酵菌株包括保加利亚乳杆菌、嗜酸乳杆菌、嗜热链球菌中的两种或三种；
A中发酵菌株接种量为2～5%（1×106～1×108cfu/mL），培养温度30～38℃，培养时间12～16h；
A中所述的增殖培养基为：胰蛋白胨10g，K2HPO4 2g，葡萄糖20g，无水醋酸钠3g，牛肉浸膏10g，柠檬酸三铵2g，酵母粉5 g，MgSO4·7H2O 0.2～0.5 g，聚山梨酯80 1mL，L-半胱氨酸0.5g，乳糖3～5g，加纯净水1000mL，充分搅匀，调整pH值在6～6.5之间；
A中控流分批补料培养中可加入补料包括玉米汁、番茄汁、麦芽汁、胡萝卜汁、平菇汁等植物成分中的一种或几种，添加流速为培养前10h低速流，10h后高速流，补料总添加量为50～100mL/L；
B中乳酸菌菌体离心条件为2500～4000r/min，离心15min；
C中冻干保护剂为：向100mL无菌蒸馏水中加入10g海藻糖，15g脱脂乳粉，5g谷氨酸钠,10g蛋白胨，混合均匀；
C中冻干保护剂与乳酸菌菌体比例为（2～2.5）：1；
D中乳酸菌菌悬液在真空冷冻干燥前需进行-80℃温度下预冻1～2h；
D中直投式果蔬汁乳酸菌促进剂活菌数含量为1×1010～1×1012cfu/g，含水量≤3%。
具体实施方式
实施例1
将保加利亚乳杆菌与嗜酸乳杆菌作为发酵菌株，按照2%浓度接种到增殖培养基中。增殖培养基配方为：胰蛋白胨10g，K2HPO4 2g，葡萄糖20g，无水醋酸钠3g，牛肉浸膏10g，柠檬酸三铵2g，酵母粉5 g，MgSO4·7H2O 0.5 g，聚山梨酯80 1mL，L-半胱氨酸0.5g，乳糖3g，加水1000mL，搅拌均匀，调整pH值至6。在37℃条件下培养12h，发酵过程中前10h慢速、后2h快速流入番茄汁50mL，以补充底物营养。将乳酸菌培养液经4000r/min转速离心15min，利用无菌蒸馏水洗涤乳酸菌菌体，重复两次。将10%海藻糖、15%脱脂乳粉、5%谷氨酸钠、10%蛋白胨混合制成的冻干保护剂溶解于无菌蒸馏水中，按照2:1比例与乳酸菌菌体制成菌悬液。将菌悬液在-80℃条件下预冻1h，再冷冻干燥，即得直投式果蔬汁乳酸菌促进剂，活菌数含量为5×1010cfu/g。
Claims (10)
1.一种直投式果蔬汁乳酸菌促进剂，其特征在于，将乳酸菌作为发酵菌株，接种至增殖培养基中在30～38℃温度下控流分批补料培养12～16h；用无菌蒸馏水离心洗涤乳酸菌菌体两次，与溶解至无菌蒸馏水的冻干保护剂相混合制成菌悬液，经低温预冻、真空冷冻干燥处理，得直投式果蔬汁乳酸菌促进剂。
2.根据权利要求1所述的发酵菌株，它包括保加利亚乳杆菌、嗜酸乳杆菌、嗜热链球菌中的两种或三种。
3.根据权利要求1所述发酵菌株接种量为2～5%（1×106～1×108cfu/mL），培养温度30～38℃，培养时间12～16h。
4.根据权利要求1所述的增殖培养基为：胰蛋白胨10g，K2HPO4 2g，葡萄糖20g，无水醋酸钠3g，牛肉浸膏10g，柠檬酸三铵2g，酵母粉5 g，MgSO4·7H2O 0.2～0.5 g，聚山梨酯80 1mL，L-半胱氨酸0.5g，乳糖3～5g，加纯净水1000mL，充分搅匀，调整pH值在6～6.5之间。
5.根据权利要求1所述控流分批补料培养中可加入补料包括玉米汁、番茄汁、麦芽汁、胡萝卜汁、平菇汁等植物成分中的一种或几种；
添加流速为培养前10h低速流，10h后为高速流；补料总添加量为50～100mL/L。
6.根据权利要求1所述乳酸菌菌体离心条件为2500～4000r/min，离心15min。
7.根据权利要求1所述冻干保护剂为：向100mL无菌蒸馏水中加入10g海藻糖，15g脱脂乳粉，5g谷氨酸钠,10g蛋白胨，混合均匀。
8.根据权利要求1所述冻干保护剂与乳酸菌菌体比例为（2～2.5）：1。
9.根据权利要求1所述真空冷冻干燥处理前应将乳酸菌菌悬液在-80℃温度下预冻1～2h。
10.根据权利要求1所述直投式果蔬汁乳酸菌促进剂活菌数含量为1×1010～1×1012cfu/g，含水量≤3%。
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