CN113604393B - 一株戊糖片球菌zf604及其应用 - Google Patents
一株戊糖片球菌zf604及其应用 Download PDFInfo
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Abstract
本发明公开了一株戊糖片球菌ZF604，其于2021年7月26日保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No:61837。所述戊糖片球菌ZF604具有高产乳酸及耐盐的特点，可在高盐发酵酱醪中正常生长，显著提高发酵原油的乳酸水平，为原油发酵提供充分的底物。将该戊糖片球菌ZF604复合现有工艺的米曲霉和酵母菌用于酱油发酵，可起到复合增香的效果，利用乳酸与原油中的醇类等风味物质进行复合反应，不仅整体提升了酱油中风味物质的含量，而且丰富了酱油中的风味物质种类，使酱油中新增了3‑甲硫基丙醇、2‑甲基‑6‑[[(E)‑1‑丙烯基]吡嗪和1,3,5,7‑环辛酸酯这些香气成分。
Description
技术领域
本发明属于微生物发酵技术领域，尤其涉及一株高产乳酸及耐盐的戊糖片球菌ZF604及其应用。
背景技术
根据发酵工艺的不同，酱油主要分为高盐稀态发酵酱油和低盐固态发酵酱油两大类。由于这两种工艺在发酵温度、盐水浓度及发酵时间等方面存在着很大的区别，所得酱油在感官品质及风味上有着明显的区别。研究表明，高盐稀态发酵酱油中香气物质的种类及相对含量要明显高于低盐固态酱油。因此，高盐稀态发酵酱油更能符合消费者的需求，也是今后生产酱油的主要发展方向。
香气特征是评价高盐稀态酱油品质的重要指标，但现有对我国高盐稀态酱油香气的研究还不够***和深入。其现代规模化酿造中多采用单一米曲霉纯培养制曲，制醪后虽也会经历蛋白质分解和乳酸菌产酸使pH快速下降的过程，但微生物的多样性不足，导致酱油的风味较为单一，风味浓郁度不足。另外，近年来对酱油香气的研究重点在于4-乙基愈创木酚和呋喃酮这些成分。然而，3-甲硫基丙醇，英文名称为1-Propanol,3-(methylthio)-，中文别名为3-(甲基硫代)-1-丙醇，分子式为C4H10OS，分子量为106.18，该物质具有强烈的圆葱、肉臭味，稀释会出现酱油气味，主要用于酱调味。本发明人经研究发现，3-甲硫基丙醇的存在可增强酱油的香气，提升酱油香气的丰富度，但目前尚未见有通过乳酸菌强化提升发酵原油中3-甲硫基丙醇含量的报道出现。
发明内容
为解决上述现有技术中存在的缺点和不足，本发明的目的在于提供一株高产乳酸及耐盐的戊糖片球菌ZF604，利用该菌株提高发酵原油的乳酸水平，为原油发酵提供充分的底物，使酱油中香气成分的种类更多，香气成分的总含量更高，酱油香气更强。
为实现其目的，本发明采取的技术方案如下：
一株戊糖片球菌ZF604(Pediococcus pentosaceus)，其于2021年7月26日保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No:61837。保藏地址为中国广州市先烈中路100号大院59号楼5楼。
所述戊糖片球菌ZF604具有高产乳酸及耐盐的特点，其菌落特征如下：菌落呈圆形，培养初期颜色呈乳白色，后期转为乳黄色，菌落不透明，但四周边缘呈透明圈。
所述戊糖片球菌ZF604为发明人对发酵酱醪进行条件性筛选后获得的，经鉴定，该菌株属于戊糖片球菌(Pediococcus pentosaceus)，将其添加至酱醪发酵中可显著提高发酵原油的乳酸含量。
本发明还提供了所述戊糖片球菌ZF604在食品发酵中的应用。在一些实施方式中，所述食品包括酱油或发酵酱。
本发明还提供了一种食品的发酵方法，其包括添加所述戊糖片球菌ZF604进行发酵。在一些实施方式中，所述食品包括酱油或发酵酱。
本发明还提供了一种提高酱油中风味物质含量的方法，其包括添加所述戊糖片球菌ZF604进行发酵。
优选地，所述方法还包括复合添加米曲霉和酵母菌进行发酵。
优选地，所述风味物质包括3-甲硫基丙醇、2-甲基-6-[[(E)-1-丙烯基]吡嗪、1,3,5,7-环辛酸酯、辛酸乙酯、壬酸乙酯、月桂酸乙酯中的至少一种。
本发明人通过研究发现，将戊糖片球菌ZF604与米曲霉和酵母菌组合应用于酱油发酵中，可起到复合增香的效果，利用乳酸与原油中的醇类等风味物质进行复合反应，不仅可使酱油中增加3-甲硫基丙醇、2-甲基-6-[[(E)-1-丙烯基]吡嗪和1,3,5,7-环辛酸酯这些新的香气成分，而且还能整体提升酱油中风味物质的含量。
本发明还提供了一种发酵菌剂，其包括所述的戊糖片球菌ZF604。
优选地，所述发酵菌剂还包括米曲霉和酵母菌。
与现有技术相比，本发明的有益效果为：本发明提供的戊糖片球菌ZF604具有高产乳酸及耐盐的特点，可在高盐发酵酱醪中正常生长，显著提高发酵原油的乳酸水平，为原油发酵提供充分的底物。将该戊糖片球菌ZF604复合现有工艺的米曲霉和酵母菌用于酱油发酵，可起到复合增香的效果，利用乳酸与原油中的醇类等风味物质进行复合反应，不仅整体提升了酱油中风味物质的含量，而且丰富了酱油中的风味物质种类，使酱油中新增了3-甲硫基丙醇、2-甲基-6-[[(E)-1-丙烯基]吡嗪和1,3,5,7-环辛酸酯这些香气成分。
附图说明
图1为戊糖片球菌ZF604菌落的形态镜检图；
图2为实施例3中组1和组3的香气组分差异分析图。
具体实施方式
为更好的说明本发明的目的、技术方案和优点，本发明通过下列实施例进一步说明。显然，下列实施例仅是本发明的一部分实施例，而不是全部的实施例。应理解，本发明实施例仅用于说明本发明的技术效果，而非用于限制本发明的保护范围。除非特别说明，否则本发明实施例中采用的方法和试验条件均为本领域常规使用的方法和试验条件，使用的试剂、设备和培养基均为本领域常规使用的试剂、设备和培养基，均可通过现有方法制备获得或通过市售购买获得。实施例中，所述百分含量如无特别说明，均为质量百分含量。实施例中使用的米曲霉为沪酿3.042，酵母菌为我司专利保藏酵母菌ZB118(广东微生物保藏中心保藏号GDMCC No:60280)。
实施例1戊糖片球菌ZF604菌株的获得
通过对生产酱醪进行条件性选育(耐盐、产酸)，筛选平板为添加2％碳酸钙的MRS培养基，筛选具备溶解圈的菌落，获得7株菌，其中5株相对对照菌株的耐盐性提升，其中4株相对对照菌株(植物乳杆菌沪酿1.08)有明显的乳酸生成量提升(于含盐豆汁液体中培养产酸活性)，综合耐盐性和乳酸生成量提升的菌株共有3株，分别为RS1703、RS1710和ZF604；发酵指标分析见表1。
表1菌株筛选及发酵乳酸质量分析结果
备注：以对照菌株值为1，其他菌株换算成相应比值。
实施例2戊糖片球菌ZF604菌株对原油乳酸含量的提升效果
将上述选育的3株高产乳酸、高耐盐的菌株与酱油主发酵菌米曲霉复合使用，进行发酵原油培养基中的效果对比，获得指标如下表2所示：
从表中可看出，3株菌种对原油的总酸含量和乳酸含量均有一定程度的提升，其中，菌株ZF604对原油乳酸含量的提升效果最优。然而，乳酸虽相对其他有机酸(醋酸、柠檬酸等)的酸味更为柔和不刺激，但总酸的过量生成也会使原油口感偏酸，不利于风味提升。为此，本发明将菌株ZF604与酵母菌和米曲霉进行复合增香，利用乳酸与原油中的醇类等风味物质进行复合反应。
表2乳酸菌菌株筛选及发酵乳酸质量分析结果
注：以对照各指标为基准，计算相对提升含量。
实施例3戊糖片球菌ZF604与酵母菌、米曲霉复合增香的效果提升
(1)将戊糖片球菌ZF604与米曲霉和/或酵母菌复合使用，进行发酵原油培养基中的效果对比，获得指标如下表3所示：
从表中可看出，单独使用米曲霉时，原油的总酸含量较高；而在米曲霉发酵中添加酵母菌后，对总酸生成有一定的抑制作用；在米曲霉发酵中添加戊糖片球菌ZF604后，总酸和乳酸的含量都会增加；在米曲霉发酵中同时添加酵母菌和戊糖片球菌ZF604后，使戊糖片球菌ZF604与酵母菌复合使用，一方面可以通过酵母菌抑制其他产酸菌的快速繁殖产生的刺激性酸，另一方面后期强化乳酸生成，有助于生乳酸、生香和呈味。
表3ZF604与酵母菌复合发酵质量分析结果
注：以对照各指标为基准，计算相对提升含量。
对组1～组3所得原油进行香气感官评价，由经验丰富的感官评定师对各试样进行鉴评描述，结果如表4所示。
表4原油香气的感官评价
(2)将发酵获得的原油采用气相色谱-质谱联用仪，分析组1和组3的香气组分。具体GC-MS分析方法如下：
设备名：安捷伦8890-5977B气相质谱联用仪
采用DB-INNOWAX 60m×0.25mm×0.25μm的气相色谱柱；
进样口温度为250℃；
升温条件为：初始温度为40℃，保持5min，以5℃每分钟升温至240℃保持15min，载气流速为1.2mL/min，不分流进样。
电子轰击电离(EI)离子源，离子源温度250℃，传输线温度280℃，质量扫描范围为30～500m/z。
结果如图2所示(图2中，米曲霉+酵母菌代表组1，米曲霉+酵母菌+ZF604代表组3)。从图2可看出，在酱油发酵过程中强化戊糖片球菌ZF604可使酱油中主要风味物质：酯类、酚类含量提升，醇类、酸类、醛减少。
(3)通过对比对照组1和组3中差异化的香气组分差异，发现对照组与实验组在不同香气物质上存在增加、降低，以及新增成分的情况，具体结果如表5所示。从表中数据可看出，添加戊糖片球菌ZF604对酱油中的酯类物质含量有一定的增加，对醋酸、糠醛、糠醇、苯乙醇、麦芽酚和棕榈酸乙酯的含量有一定的减少，同时还使酱油中增强了3-甲硫基丙醇、2-甲基-6-[[(E)-1-丙烯基]吡嗪和1,3,5,7-环辛酸酯这些新的香气成分。其中，3-甲硫基丙醇的含量较高，3-甲硫基丙醇的英文名称为1-Propanol,3-(methylthio)-，中文别名为3-(甲基硫代)-1-丙醇，分子式为C4H10OS，分子量为106.18，该物质有强烈的圆葱和肉臭味，稀释会出现酱油气味，目前主要用于酱调味，其存在可增强酱油香气。
表5对照样与试验样香气部分关键物质成分变化
注：表中“-”表示不含该成分。其他组分未全部列出。
最后应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。
Claims (8)
1.一株戊糖片球菌ZF604，其于2021年7月26日保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No:61837，保藏地址为中国广州市先烈中路100号大院59号楼5楼。
2.如权利要求1所述的戊糖片球菌ZF604在酱油或发酵酱发酵中的应用。
3.一种食品的发酵方法，其特征在于，包括添加权利要求1所述的戊糖片球菌ZF604进行发酵。
4.如权利要求3所述的发酵方法，其特征在于，所述食品为酱油或发酵酱。
5.一种发酵菌剂，其特征在于，包括权利要求1所述的戊糖片球菌ZF604。
6.如权利要求5所述的发酵菌剂，其特征在于，还包括米曲霉和酵母菌。
7.一种提高酱油中风味物质含量的方法，其特征在于，包括添加权利要求1所述的戊糖片球菌ZF604、米曲霉和酵母菌进行发酵。
8.如权利要求7所述的方法，其特征在于，所述风味物质包括3-甲硫基丙醇、2-甲基-6-[[(E)-1-丙烯基]吡嗪、1,3,5,7-环辛酸酯、辛酸乙酯、壬酸乙酯、月桂酸乙酯中的至少一种。
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