CN111944731B - 一种基于多菌共酵技术改善发酵果蔬风味的方法 - Google Patents
一种基于多菌共酵技术改善发酵果蔬风味的方法 Download PDFInfo
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Abstract
本发明公开了一株植物乳杆菌(Lactobacillus plantarum)J68,保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCC No.20193。本发明还公开了一种利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,包括步骤:将植物乳杆菌J68菌液和戊糖片球菌C53菌液接种在果蔬发酵体系中进行发酵,所述发酵体系的液体中植物乳杆菌J68和戊糖片球菌C53的菌株终浓度均为106~108CFU/mL。使用本发明所述方法制备酸菜,能够显著提高总酸和乳酸的含量,提高酸菜中氨基酸的含量,改善酸菜的风味和滋味。
Description
技术领域
本发明涉及微生物发酵剂技术领域,具体涉及植物乳杆菌(Lactobacillusplantarum)和戊糖片球菌(Pediococcus pentosaceus)共酵在改善发酵果蔬风味品质的应用。
背景技术
酸菜是一种在东北地区非常受欢迎的发酵食品。在冬季,基本上每家每户都会大量的腌制酸菜,大多都是采用传统的腌制方式,首先将白菜和盐叠放进酸菜缸中,加水没过白菜,然后密封或者用石头压住,一个月之后发酵完成。目前,市售酸菜主要出自小作坊,采用传统工艺腌制而成,产品酸咸十足。但是产品品质不稳定,易发生腐败等。
接种发酵能够提高产品的品质稳定性,缩短发酵周期。乳酸菌作为发酵蔬菜中的优势菌,常常被作为发酵剂,应用于发酵制品的工艺改进和品质改良。酸菜发酵是多种微生物相互作用的结果。目前,单菌株接种发酵常常被用于酸菜的品质控制,但单菌株发酵酸菜的风味品质不及自然发酵浓郁。
发明内容
本发明的目的在于提供一种利用植物乳杆菌和戊糖片球菌共酵改善发酵果蔬风味品质的方法。
为达到上述目的,本发明提供了一株植物乳杆菌(Lactobacillus plantarum)J68,保藏号为CGMCC No.20193,于2020年07月06日保藏在中国微生物菌种保藏管理委员会普通微生物中心,简称CGMCC,地址为:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所。
本发明还提供了一种戊糖片球菌(Pediococcus pentosaceus)C53,保藏号为CGMCC No.20192,于2020年07月06日保藏于“中国微生物菌种保藏管理委员会普通微生物中心”,简称CGMCC,地址为:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所。
所述植物乳杆菌J68与戊糖片球菌C53分离自酸菜样品,采用细菌通用引物27F/1492R进行16S rDNA序列鉴定,为植物乳杆菌和戊糖片球菌。
本发明同时提供了所述植物乳杆菌J68和戊糖片球菌C53在改善发酵果蔬风味品质中的应用。
优选方式下,所述发酵果蔬为以白菜为原料发酵所得的酸菜;所述改善发酵果蔬风味包括,增加发酵果蔬中氨基酸的含量、增加有机酸的含量,和增加挥发性风味物质的含量;所述挥发性风味物质为酯类、醛类、烃类和腈类化合物,所述有机酸为酒石酸、苹果酸、乙酸、柠檬酸、琥珀酸。
一种利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,包括步骤:
将菌株浓度为108~1010CFU/mL的植物乳杆菌J68与菌株浓度为108~1010CFU/mL的戊糖片球菌C53作为发酵剂,接种至含有果蔬的发酵体系内,使得发酵体系的液体中植物乳杆菌J68菌株接种终浓度为106~108CFU/mL,戊糖片球菌C53菌株接种终浓度为106~108CFU/mL,所述植物乳杆菌J68与戊糖片球菌C53在发酵体系的液体中的菌落数终浓度比为(1~3):1,进行发酵;其中,所述果蔬与所述液体的重量体积比是1:(2~4)g/mL;所述植物乳杆菌J68的保藏号为CGMCC No.20193,所述戊糖片球菌C53的保藏号为CGMCCNo.20192。
优选方式下,所述发酵果蔬为酸菜,所述果蔬为白菜。
优选方式下,所述利用所述植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,包括步骤:
S1、原料预处理:
新鲜的白菜,置于自来水煮沸,放凉备用,得待腌制白菜;其中,可以根据容器的大小和实际生产需要将白菜切分成适当的大小;
S2、制作酸菜:
将NaCl溶解在水中,制备成NaCl溶液;将所述NaCl溶液与植物乳杆菌J68菌液和戊糖片球菌C53菌液混合,制得腌制液,所述腌制液中植物乳杆菌J68菌株接种终浓度为106~108CFU/mL,戊糖片球菌C53菌株接种终浓度为106~108CFU/mL,所述植物乳杆菌J68与戊糖片球菌C53在发酵体系的液体中的菌落数终浓度比为(1~3):1;将所述腌制液与步骤S1所述待腌制白菜置于灭菌容器中,14~16℃水封放置发酵28~32天,得酸菜;
其中,所述NaCl溶液中,NaCl和水的重量体积比是1:45~47g/mL;所述待腌制白菜与所述腌制液重量体积比是1:(2~4)g/mL。
优选方式下,所述植物乳杆菌J68菌液或戊糖片球菌C53菌液的制备方法为:挑取平板上的单菌落,接种至2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液A;然后取10μL所述菌液A接种至50mL MRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液B;将所述菌液B置于10000r/min离心10min后收集菌体,将菌体用无菌生理盐水(质量分数0.85%氯化钠水溶液)稀释成108~1010CFU/mL的菌悬液;
所述MRS肉汤培养基成分为:蛋白胨10.0g/L、牛肉浸粉8.0g/L、酵母浸粉4.0g/L、葡萄糖20.0g/L、磷酸氢二钾2.0g/L、柠檬酸氢二铵2.0g/L、乙酸钠5g/L、硫酸镁0.2g/L、硫酸锰0.04g/L、吐温80 1.0g/L,25℃的pH值为5.7±0.2。
优选方式下,所述灭菌容器的灭菌方法为:将容器洗净,置于沸水中加热灭菌20min。
优选方式下,所述利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,包括步骤:
S1、原料预处理:取白菜,每个白菜按重量平均切份成四份,用自来水煮沸,放凉备用,得待腌制白菜;
S2、制作酸菜:
将容器清洗后,沸水灭菌20min,得灭菌容器;
将36g食盐溶解于1680mL水中并与8mL 1.4×109CFU/mL的植物乳杆菌J68菌液和2mL 2.3×109CFU/mL的戊糖片球菌C53菌液混合,制得腌制液;取步骤S1所述待腌制白菜600g置于所述灭菌容器中,加入所述腌制液,15℃水封放置发酵30天,得酸菜;
其中,所述植物乳杆菌J68菌液制备方法为:挑取植物乳杆菌J68菌落,接种于2mLMRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液A,随后取10μL所述菌液A接种至50mLMRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液B;将所述菌液B置于10000r/min离心10min后收集菌体,将菌体用用质量分数0.85%氯化钠水溶液稀释成1.4×109CFU/mL的菌悬液;
所述戊糖片球菌C53菌液的制备方法为:挑取戊糖片球菌C53单菌落,接种于2mLMRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液C,随后取10μL所述菌液C接种至50mLMRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液D;将所述菌液D置于10000r/min离心10min后收集菌体,将菌体用用质量分数0.85%氯化钠水溶液稀释成成2.3×109CFU/mL的菌悬液。
本发明的有益效果是:
多菌共酵可以在一定程度上模拟酸菜的自然发酵微生态,进而改善单菌接种酸菜的风味品质。
将植物乳杆菌J68和戊糖片球菌C53应用于酸菜发酵,共酵能够显著提高酸菜中总酸和乳酸的含量,提高氨基酸的含量。另外,二者共酵还能改善酸菜风味,提高酯类、醛类、烃类和腈类化合物含量。还能改善酸菜滋味,使共酵酸菜风味和滋味更接近于自然发酵。
本发明提供的植物乳杆菌和戊糖片球菌,可用改善发酵果蔬尤其是酸菜风味品质,具有非常广泛的应用前景。通过接种乳酸菌发酵酸菜能够降低酸菜的亚硝酸盐含量,缩短发酵时间,提高生产效率,并对酸菜的氨基酸含量和挥发性风味成分等品质指标均有影响。
附图说明
图1为不同pH条件下植物乳杆菌J68的生长曲线;
图2为不同pH条件下戊糖片球菌C53的生长曲线
图3为自然发酵和多菌共酵酸菜的理化性质;
图4为自然发酵和多菌共酵酸菜的挥发性风味物质含量。
具体实施方式
下面通过具体实施实例对本发明做进一步说明。
本发明提供了两株菌株—植物乳杆菌(Lactobacillus plantarum)J68和戊糖片球菌(Pediococcus pentosaceus)C53,保藏在中国微生物菌种保藏管理委员会普通微生物中心(地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所),分类命名为植物乳杆菌(Lactobacillus plantarum)和戊糖片球菌(Pediococcus pentosaceus),保藏号分别为CGMCC No.20193和CGMCC No.20192。
上述植物乳杆菌J68与戊糖片球菌C53来自发酵食品,采用细菌通用引物27F/1492R进行16S rDNA序列鉴定,为植物乳杆菌和戊糖片球菌。
所述菌株的保藏方法:从MRS琼脂培养基平板上挑选生长良好的单菌落接种至2mLMRS肉汤培养基,37℃、耗氧、静置培养24h,取1mL菌液加入到含有1mL体积分数30%甘油的冻存管中,于-80℃冰箱保藏。
所述菌株的培养方法:从MRS琼脂培养基平板上挑选生长良好的单菌落接种至2mLMRS肉汤培养基中,37℃、耗氧、静置培养24h,随后取10Μl菌液接种至50mL MRS肉汤培养基中,37℃、耗氧、静置培养12h,10000r/min离心10min后收集菌体,将菌体用质量分数为氯化钠0.85%水溶液稀释,制备成108~1010CFU/mL的菌悬液。
所述MRS肉汤培养基成分为:蛋白胨10.0g/L、牛肉浸粉8.0g/L、酵母浸粉4.0g/L、葡萄糖20.0g/L、磷酸氢二钾2.0g/L、柠檬酸氢二铵2.0g/L、乙酸钠5g/L、硫酸镁0.2g/L、硫酸锰0.04g/L、吐温80 1.0g/L,25℃的pH值为5.7±0.2。
为表征上述菌株的耐酸特性,采用全自动生长曲线分析仪(芬兰,FP-1100-C)测定菌株J68和C53在不同pH条件下的生长情况,具体步骤如下:挑取单菌落接种至2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,随后取10μL菌液接种至300μL MRS肉汤培养基中,然后放入全自动生长曲线分析仪进行测定。结果如图1和图2所示。由图1可以看出菌株J68在2h后进入指数生长期,12h后进入稳定期。菌株C53在2h后进入指数增长期,10h后进入稳定期。在pH=4条件下,J68和C53仍然可以继续生长,表明在酸化过程中仍可以进行物质代谢,促进风味的产生。
本发明同时提供了一种利用所述植物乳杆菌和戊糖片球菌发酵改善风味品质的方法:将菌株浓度为108~1010CFU/mL的植物乳杆菌J68与戊糖片球菌C53作为发酵剂,按浓度比为(1~3):1接种至发酵体系内,使发酵体系中菌株浓度达到106~108CFU/mL,进行发酵;其中,所述植物乳杆菌J68和戊糖片球菌C53的保藏号为CGMCC No.20193和CGMCCNo.20192;
利用植物乳杆菌J68和戊糖片球菌C53应用于酸菜发酵,二者共酵能够显著提高酸菜中总酸和有机酸的含量,还能够提高氨基酸的含量。另外,二者共酵还能改善酸菜风味,提高酯类、醛类、烃类和腈类化合物含量。改善酸菜滋味,使共酵酸菜风味和滋味更接近于自然发酵。
实施例1:
一种利用植物乳杆菌J68和戊糖片球菌C53改善酸菜风味品质的方法,包括步骤:
S1、原料预处理:
新鲜的白菜,挑选出有杂质、虫眼、腐烂的菜叶丢弃,一个按重量平均切成四份备用;
自来水煮沸,放凉备用,得待腌制白菜;
S2、制作酸菜:
将容器清洗后,沸水灭菌20min,得灭菌容器;将36g食盐溶解于1680mL水中并与8mL 1.4×109CFU/mL的植物乳杆菌J68菌液和2mL 2.3×109CFU/mL的戊糖片球菌C53菌液混合,制得腌制液,所述腌制液中植物乳杆菌J68浓度为0.66×107CFU/mL,戊糖片球菌C53浓度为0.27×107CFU/mL;取步骤S1所得600g待腌制白菜置于所述灭菌容器中,加入所述腌制液,15℃水封放置发酵30天,得酸菜;
所述植物乳杆菌J68菌液制备方法为:挑取植物乳杆菌J68单菌落,接种于2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液A,随后取10μL所述菌液A接种至50mL MRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液B;将所述菌液B置于10000r/min离心10min后收集菌体,将菌体用无菌生理盐水(质量分数0.85%氯化钠水溶液)制备成1.4×109CFU/mL的菌悬液;
所述戊糖片球菌C53菌液的制备方法为:挑取戊糖片球菌C53单菌落,接种于2mLMRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液C,随后取10μL所述菌液C接种至50mLMRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液D;将所述菌液D置于10000r/min离心10min后收集菌体,将菌体用用质量分数0.85%氯化钠水溶液稀释成成2.3×109CFU/mL的菌悬液。
所述MRS肉汤培养基成分为:蛋白胨10.0g/L、牛肉浸粉8.0g/L、酵母浸粉4.0g/L、葡萄糖20.0g/L、磷酸氢二钾2.0g/L、柠檬酸氢二铵2.0g/L、乙酸钠5g/L、硫酸镁0.2g/L、硫酸锰0.04g/L、吐温80 1.0g/L,25℃的pH值为5.7±0.2。
对比例1:
自然发酵制备酸菜步骤:
S1、原料预处理:
新鲜的白菜,挑选出有杂质、虫眼、腐烂的菜叶丢弃,一个切成四份备用;
自来水煮沸,放凉备用;
S2、制作酸菜:
将容器清洗后,沸水灭菌20min;取步骤S1所得600g白菜块置于所述容器中,将36g食盐溶解于1680ml水中倒入放置白菜的容器中,15℃水封放置发酵30天,得酸菜。
对比例2:
一种利用植物乳杆菌J68改善酸菜风味品质的方法,包括步骤:
S1、原料预处理:
新鲜的白菜,挑选出有杂质、虫眼、腐烂的菜叶丢弃,一个切成四份备用;
自来水煮沸,放凉备用;
S2、制作酸菜:
将容器清洗干净,沸水灭菌20min;取步骤S1所得600g白菜块置于所述容器中,将36g食盐溶解于1680mL水中并与12mL浓度为1.4×109CFU/mL的植物乳杆菌J68菌体混合,食盐水和菌体混合后倒入放置白菜的容器中,使得腌制液中植物乳杆菌J68浓度为0.99×107CFU/mL,15℃水封放置发酵30天,得酸菜;
其中,所述植物乳杆菌J68菌液的制备方法为:挑取J68单菌落,接种于2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,取10μL菌液接种至MRS肉汤培养基中,37℃、耗氧、静置培养12h;10000r/min离心10min后收集菌体,将菌体用无菌生理盐水(0.85%氯化钠水溶液,w/v,g/mL)制备成1.4×109CFU/mL的菌悬液。
对本发明实施例及对比例制备的酸菜的游离氨基酸、电子舌、挥发性风味物质进行了测定。
一、理化性质
利用pH计、酸碱滴定法、盐酸萘乙二胺法和离子色谱法测定酸菜样品的pH值、总酸、亚硝酸盐和乳酸含量,结果如图3所示。植物乳杆菌J68和戊糖片球菌C53共酵能够显著提高酸菜的总酸和乳酸含量。
二、游离氨基酸含量
运用氨基酸测定仪LA8080测定酸菜中的氨基酸含量,结果如表2所示。与对比例1相比,除了天冬氨酸,半胱氨酸,甲硫氨酸有略微下降,对比例2和实施例1中酸菜的游离氨基酸含量均有较明显的增长,其中苏氨酸、丝氨酸、谷氨酸、脯氨酸、甘氨酸、丙氨酸、蛋氨酸、异亮氨酸增长幅度最为明显。表明接种发酵能够增加氨基酸的含量,改善产品的滋味。
表1酸菜中游离氨基酸含量(mg/L)
三、挥发性风味物质的测定
采用顶空固相微萃取技术结合GC/MS对酸菜中的挥发性化合物进行分析,结果如表3和图4所示。与对比例1酸菜相比,对比例2和实施例1酸菜中酯类化合物含量显著增加,表明接种发酵能够增加酯类化合物的含量,而实施例1酸菜中的酯类化合物的含量高于对比例1和对比例2,表明共酵有助于酯类化合物的产生。而酯类物质是发酵食品中的主要呈香化合物。另外,J68和C53共酵还能够显著提高醛类化合物,而醛类化合物的阈值较低,香气较好,对酸菜的风味物质形成贡献大。除此之外,还能提高烃类和醚类化合物的含量,构成酸菜挥发性风味成分,醚类茴香脑具有茴香香气,可为酸菜增香。减少蔬菜中固有的异硫氰酸类化合物含量,能够减少白菜本身的刺激性辛辣风味。
表2酸菜中挥发性风味物质含量(mg/L)
三、有机酸
采用液相色谱分析分析对比例1、对比例2和实施例1所制备酸菜的有机酸含量,结果如表3所示。与对比例1相比,对比例2和实施例1中各有机酸的含量显著提高,表明接种发酵能够提高酸菜中有机酸的含量。与对比例2相比,实施例1酸菜中的有机酸含量显著升高,表明J68和C53共酵能够有助于有机酸的提高,增加酸菜的酸味。
表4酸菜中有机酸含量(g/L)
四、电子舌测定
采用TS-5000Z型味觉分析系统(Insent,Japan)对本发明实施例和对比例所得酸菜的风进行测定,结果如表3所示。与对比例1酸菜相比,接种发酵的酸味显著提高。酸味是酸菜中重要的感官评价指标,对比例1酸菜的酸值最低,对比例2单菌接种发酵酸菜中酸值最高。但过高的酸味会影响酸菜的感官,而实施例1两株菌共酵酸菜的酸味位于二者之间。另外,接种发酵能够减少酸菜的苦味和鲜味。因此,接种发酵能改善酸菜的滋味品质。
表4电子舌统计结果
a , b,c,d不同上标字母在行内差异有统计学意义(p<0.05)。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明披露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (9)
1.一种植物乳杆菌J68和戊糖片球菌C53在改善发酵果蔬风味品质的中的应用,其特征在于:
所述植物乳杆菌Lactobacillus plantarum J68,保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCC No.20193;
所述戊糖片球菌Pediococcus pentosaceus C53,保藏于“中国微生物菌种保藏管理委员会普通微生物中心”,保藏号为CGMCC No.20192。
2.根据权利要求1所述戊糖片球菌J68和戊糖片球菌C53在改善发酵果蔬风味品质中的应用,其特征在于,所述发酵果蔬为以白菜为原料发酵所得的酸菜。
3.根据权利要2所述戊糖片球菌J68和戊糖片球菌C53在改善发酵果蔬风味品质中的应用,其特征在于,所述改善发酵果蔬风味包括,增加发酵果蔬中氨基酸的含量、增加有机酸的含量,增加挥发性风味物质的含量。
4.一种利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,包括步骤:
将菌株浓度为108~1010CFU/mL的植物乳杆菌J68与菌株浓度为108~1010CFU/mL的戊糖片球菌C53作为发酵剂,接种至含有果蔬的发酵体系内,进行发酵;其中,所述发酵体系的液体中植物乳杆菌J68菌株终浓度为106~108CFU/mL,戊糖片球菌C53菌株终浓度为106~108CFU/mL;所述植物乳杆菌J68与所述戊糖片球菌C53在发酵体系的液体中的菌落数终浓度比为(1~3):1;所述果蔬与所述液体的重量体积比是1:(2~4)g/mL;所述植物乳杆菌J68的保藏号为CGMCC No.20193,所述戊糖片球菌C53的保藏号为CGMCC No.20192。
5.根据权利要求4所述利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,所述果蔬为白菜。
6.根据权利要求5所述利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,包括步骤:
S1、原料预处理:取白菜,置于水中煮沸,放凉备用,得待腌制白菜;
S2、制作酸菜:将NaCl溶解在水中,制备成NaCl溶液;将所述NaCl溶液与植物乳杆菌J68菌液和戊糖片球菌C53菌液混合,制得腌制液,将所述腌制液与步骤S1所述待腌制白菜置于灭菌容器中,14~16℃水封发酵28~32天,得酸菜;
其中,所述NaCl溶液中,NaCl和水的重量体积比是1:45~47g/mL;所述腌制液中植物乳杆菌J68菌株终浓度为106~108CFU/mL,戊糖片球菌C53菌株终浓度为106~108CFU/mL;所述植物乳杆菌J68与戊糖片球菌C53在发酵体系的液体中的菌落数终浓度比为(1~3):1;所述待腌制白菜与所述腌制液重量体积比是1:(2~4)g/mL。
7.根据权利要求6所述利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,所述植物乳杆菌J68菌液或戊糖片球菌C53菌液的制备方法为:挑取平板上的单菌落,接种至2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液A;然后取10μL所述菌液A接种至50mL MRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液B;将所述菌液B置于10000r/min离心10min后收集菌体,将菌体用质量分数0.85%氯化钠水溶液稀释成108~1010CFU/mL的菌悬液。
8.根据权利要求6所述利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,所述灭菌容器的灭菌方法为:将容器洗净,置于沸水中加热灭菌20min。
9.根据权利要求6所述利用植物乳杆菌J68和戊糖片球菌C53改善发酵果蔬风味品质的方法,其特征在于,包括步骤:
S1、原料预处理:取白菜,每个白菜按重量平均切份成四份,用自来水煮沸,放凉备用,得待腌制白菜;
S2、制作酸菜:将容器清洗后,沸水灭菌20min,得灭菌容器;将36g食盐溶解于1680mL水中并与8mL 1.4×109CFU/mL的植物乳杆菌J68菌液和2mL2.3×109CFU/mL的戊糖片球菌C53菌液混合,制得腌制液;取步骤S1所述待腌制白菜600g置于所述灭菌容器中,加入所述腌制液,15℃水封放置发酵30天,得酸菜;
其中,所述植物乳杆菌J68菌液制备方法为:挑取植物乳杆菌J68菌落,接种于2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液A,随后取10μL所述菌液A接种至50mL MRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液B;将所述菌液B置于10000r/min离心10min后收集菌体,将菌体用质量分数0.85%氯化钠水溶液稀释成1.4×109CFU/mL的菌悬液;
所述戊糖片球菌C53菌液的制备方法为:挑取戊糖片球菌C53单菌落,接种于2mL MRS肉汤培养基中,37℃、耗氧、静置培养24h,得菌液C,随后取10μL所述菌液C接种至50mL MRS肉汤培养基中,37℃、耗氧、静置培养12h,得菌液D;将所述菌液D置于10000r/min离心10min后收集菌体,将菌体用质量分数0.85%氯化钠水溶液稀释成成2.3×109CFU/mL的菌悬液。
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