CN114107099A - 一株能够提升发酵香肠风味品质的植物乳杆菌及应用 - Google Patents
一株能够提升发酵香肠风味品质的植物乳杆菌及应用 Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
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- A23B4/22—Microorganisms; Enzymes; Antibiotics
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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
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
- A23L13/45—Addition of, or treatment with, microorganisms
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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
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
- A23L13/65—Sausages
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- 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
- C12N1/20—Bacteria; Culture media therefor
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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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
本发明提供了一株能够提升发酵香肠风味品质的植物乳杆菌。本发明中的植物乳杆菌保藏在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),保藏编号为:CGMCC No.22506,保藏日期为2021年05月06日,分类命名为植物乳杆菌(Lactobacillus plantarum)。该菌株分离自黑龙江省牡丹江市的传统风干肠,其发酵特性能够满足肉类发酵剂的要求,适用于肉类发酵。该菌株能够提升发酵香肠中挥发性化合物的含量,降低体系的pH,从而提升北方传统风干肠的风味和安全特性。
Description
技术领域
本发明属于微生物技术领域,涉及一株从风干肠中筛选出的发酵特性良好的植物乳杆菌 MDJ2(Lactobacillus plantarum MDJ2)及其在发酵香肠中的应用。
背景技术
风干肠是我国典型的北方传统发酵肉制品,因其独特的口感和风味而深受消费者的喜爱。通常,风干肠主要由绞碎的瘦肉和猪脂肪制成,与食盐、亚硝酸盐、酒、糖和香辛料等混合均匀,然后灌入天然的肠衣中。在自然或人工控制条件下,经过微生物发酵而成的发酵粉肉制品。自然发酵的过程显著增加了微生物多样性,但由于环境、气候、设备和发酵剂的不同,微生物差异较大,发酵的安全性和最终产品的品质难以控制。因此,不同批次的传统风干肠品质的一致性和安全性是亟需解决的热点问题。
合适发酵剂的使用可能是解决上述问题的有效途径之一。来源于发酵肉制品的原生发酵剂能够更好地适应肉类环境。传统发酵香肠的微生物菌群主要由乳酸菌、凝固酶阴性葡萄球菌、酵母和霉菌组成,其中乳酸菌是在肉类发酵和保藏中起主要作用,也通常作为发酵剂在发酵肉制品中使用。接种乳酸菌可迅速降低发酵肉制品的pH,缩短发酵周期、改善组织结构,促进大分子物质的代谢,产生小分子物质,包括滋味物质、风味物质前体和挥发性风味物质,从而促进产品风味的形成,同时可抑制腐败微生物的生长,保证了发酵肉制品的安全性。因此,乳酸菌的筛选和接种研究对发酵肉制品的开发有着十分重要的意义。
风味是消费者评价发酵肉制品的一个重要指标。风味物质及其前体物质通常是通过发酵过程中蛋白质和脂质的降解和代谢产生的,与微生物活动密切相关。乳酸菌能够促进蛋白质降解,蛋白质降解会产生多肽和游离氨基酸等前体物质,某些游离氨基酸通过微生物的转氨、脱氨和脱羧作用生成醛类、醇类、酯类和其他挥发性风味物质,从而促进产品的风味品质。基于此,乳酸菌接种发酵香肠可有效地提升发酵肉制品的风味,以期为发酵香肠的开发及品质提升提供理论依据。
发明内容
本发明针对以上问题,提供了一株植物乳杆菌MDJ2及其在发酵香肠中的应用,抑制腐败微生物的生长,提升香肠的风味品质。
本发明的植物乳杆菌MDJ2(Lactobacillus plantarum MDJ2)为从发酵酸乳中提取并分离鉴定的菌种,保藏在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),地址:北京市朝阳区北辰西路1号院中科院微生物研究所,其菌种保藏编号为:CGMCCNo.22506,保藏日期为2021年05月06日,菌种的分类命名为植物乳杆菌(Lactobacillusplantarum)。
本发明中的植物乳杆菌MDJ2具有以下生物学特性:
1、形态特征:植物乳杆菌MDJ2革兰氏染色结果为阳性,杆状,成短链状排列,无芽胞,无荚膜,无鞭毛。
2、菌落特征:植物乳杆菌MDJ2在MRS固体培养基上生长良好,菌落形态为白色、凸起、圆形,边缘光滑细密,直径约2-3mm。在液体培养基中均匀混浊生长。
保藏信息:
保藏单位全称:中国微生物菌种保藏管理委员会普通微生物中心;
保藏单位简称:CGMCC;
保藏单位地址:北京市朝阳区北辰西路1号院中科院微生物研究所;
保藏日期:CGMCC No.22506;
保藏日期:2021年05月06日。
具体实施方式
下面对本发明的技术方案作进一步的说明,但并不局限于此,凡是对本发明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精神和范围,均应涵盖在本发明的保护范围中。
实施例1:植物乳杆菌的培养和制备
将发酵特性良好的植物乳杆菌MDJ2接种到MRS肉汤培养基中进行发酵培养,37℃培养18h。发酵液于4℃条件下,10000×g离心10min,收集沉淀并使用无菌生理盐水洗涤两次。将制备好的发酵剂以107CFU/g接种于发酵香肠中。
实施例2:发酵香肠的制作
将瘦肉和肥肉用孔径为1.5cm筛板的绞肉机绞碎,瘦肉(猪臀肉)和肥肉(猪背脂)质量比为9:1。然后,添加0.01%亚硝酸盐(以瘦肉质量计)、1%玉泉大曲、1%葡萄糖、0.3%味素、5%水和0.8%混合香辛料(包括:花椒、橘皮、砂仁、小茴香和丁香等)。随后灌入猪小肠衣,风干肠长度约15cm,直径约2.5cm。将制备的发酵剂接种到香肠中为试验组,以未接种处理组作为对照组进行相同条件的发酵。然后在温度为25±2℃,相对湿度为 30%-50%的环境中风干1天,随后转移到恒温恒湿发酵箱中发酵11天,温度为25±2℃,相对湿度为65%-70%。测定香肠在发酵过程中的乳酸菌数和pH变化情况,并对发酵末期的游离氨基酸含量,挥发性化合物含量和感官评价的进行评估。
实施例3:发酵香肠中乳酸菌数和pH的测定
将25g香肠样品在无菌条件下剪碎,溶于225mL无菌生理盐水混匀,4℃下平衡30min。采用MRS固体培养基进行平板涂布,在30℃下培养48小时测定乳酸菌数(log CFU/g)。将试验组和对照组的香肠分别绞碎后,准确称取10.00g样品,加入90mL蒸馏水,均质后过滤,取上清液用pH计测定。
表1发酵香肠在发酵过程中乳酸菌数和pH的变化
注:同行中不同小写字母表示样品差异显著(P<0.05)。
从表1可以看出,在整个发酵周期中,与对照组香肠相比,接种的植物乳杆菌在发酵香肠中能够显著地增殖和生长。此外,接种的植物乳杆菌能够显著地降低香肠的pH,抑制有害菌的生长,延长产品的保质期,提高了产品的安全性。
实施例4:发酵香肠中游离氨基酸的测定
香肠样品(10.0g)于0.1mol/L HCl 1:5(w/v)均质30min,然后在0℃下10000g离心20min,取上清液通过0.45μm的滤膜过滤。然后,将1.0mL液体转移到玻璃管并使用异硫氰酸苯酯(PITC)试剂进行衍生化。使用配备Accucore C18柱(3×150mm,2.6μm)的高效液相色谱系统分析氨基酸,检测波长为254nm,流速为1mL/min。通过比较每种氨基酸标准品的保留时间和峰面积来定性和定量风干肠中的每种氨基酸。游离氨基酸的含量以mg/g香肠表示。
表2发酵香肠在发酵末期的游离氨基酸含量(mg/g)
注:同行中不同小写字母表示样品差异显著(P<0.05)。
n.d.:未检出。
在发酵肉制品中,游离氨基酸来源于蛋白质的降解,也是风味化合物的重要前体物质。从表2可以看出,发酵香肠中接种植物乳杆菌降低了游离氨基酸的含量,可能是由于植物乳杆菌的作用将香肠中的氨基酸转化成风味物质,促进产品风味的形成。
实施例5:发酵香肠中发性化合物含量的测定
采用顶空固相微萃取的方法提取风干肠中的挥发性化合物。将切碎的风干肠(3.0g)放入20mL顶空样品瓶中,加入4μL邻二氯苯作为内标物。在45℃条件下用50/30μmDVB/CAR/PDMS萃取头顶空吸附30min,然后立即将萃取头插入气质进样口,解析3min。
挥发性化合物的测定使用气相色谱-质谱(GC-MS)系统(QP2020,岛津),气相色谱条件:InertCap WaX毛细管柱(60m×0.25mm×0.25μm);进样口温度为230℃;不分流模式进样;以氦气为载体,流速为1mL/min;程序升温:柱初温40℃,保持3min,以5℃/min 升至200℃,再以10℃/min升至230℃,保持2min。质谱条件:离子源温度230℃;质量扫描范围45-500(m/z)。
挥发性化合物的鉴定是通过对比NIST 17质谱库,取相似度大于90%作为鉴定结果,并且通过计算标准烷烃(C6-C20)的保留指数(LRI),与文献报道结果相比较进行辅助鉴定。挥发性化合物的定量采用内标法,以溶于正己烷的邻二氯苯溶液作为内标物质,定量结果以μg/kg表示。
表3发酵香肠在发酵末期的挥发性化合物含量(μg/kg)
注:同行中不同小写字母表示样品差异显著(P<0.05)。
从表3可以看出,与对照组相比,植物乳杆菌的接种能够显著增加发酵香肠中的醛类、酮类、醇类、酸类和酯类物质的含量。因此,植物乳杆菌MDJ2的接种对发酵香肠的风味具有提升作用。
实施例6:感官评价
为了评估样品之间的感官差异,对发酵末期(第12天)的香肠进行感官评价。由9位训练有素的成员(4位男性和5位女性)组成的感官小组对熟制的发酵香肠(在90℃下15min)进行评估。该小组由以前在肌肉食品感官评估方面受过训练的实验室中的教职员工和研究生组成。向每个小组成员供应纯净水以在治疗之间清除其味觉。将香肠切片(3mm厚)并标出编号为三位随机数组成。在每次小组讨论之前,发送“热身”样本以提醒小组成员每种感官特征的得分范围。评估的指标包括:颜色,硬度,气味(发酵味、酒香味、香辛料味、脂肪味、花香味、果香味、酸败味和整体气味)。属性的强度或程度用从1(低强度)到7(高强度)的七点描述量表来表示。
表4发酵香肠在发酵末期的感官评价
感官指标 | 对照组 | 试验组 |
硬度 | 5.22±0.14<sup>b</sup> | 5.75±0.11<sup>a</sup> |
颜色 | 5.84±0.10<sup>a</sup> | 5.60±0.06<sup>a</sup> |
发酵味 | 4.39±0.01<sup>b</sup> | 4.52±0.02<sup>a</sup> |
酒香味 | 3.52±0.02<sup>a</sup> | 3.48±0.02<sup>a</sup> |
香辛料味 | 4.64±0.05<sup>a</sup> | 4.56±0.06<sup>a</sup> |
脂肪味 | 3.28±0.02<sup>a</sup> | 3.04±0.04<sup>b</sup> |
花香味 | 3.04±0.02<sup>a</sup> | 3.04±0.02<sup>a</sup> |
果香味 | 3.41±0.02<sup>a</sup> | 3.43±0.02<sup>a</sup> |
酸败味 | 1.13±0.02<sup>a</sup> | 1.11±0.02<sup>a</sup> |
整体气味 | 5.52±0.02<sup>b</sup> | 5.67±0.03<sup>a</sup> |
注:同行中不同小写字母表示样品差异显著(P<0.05)。
从表4可知,对照组与试验组香肠样品的颜色、酒香味、香辛料味、花香味、果香味、酸败味的强度无显著性差异(P>0.05)。试验组香肠的硬度、发酵味、整体气味的强度比对照组强(P<0.05),而对照组的脂肪味的强度比试验组强。综上可知,接种植物乳杆菌MDJ2 能够提升发酵香肠的整体气味并抑制脂肪氧化产生的脂肪味。
Claims (2)
1.一株能够提升发酵香肠风味的植物乳杆菌MDJ2(Lactobacillus plantarum MDJ2),保藏于中国微生物菌种保藏管理委员会普通微生物中心,菌种保藏编号为:CGMCCNo.22506。
2.权利要求1所述的能够提升发酵香肠风味的植物乳杆菌MDJ2,可应用在发酵肉制品中作为发酵剂。
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