CN117106620A - 一株植物乳植杆菌及其应用 - Google Patents
一株植物乳植杆菌及其应用 Download PDFInfo
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Abstract
本发明公开了一株植物乳植杆菌及其应用,属于微生物及食品领域,该菌株保藏于中国典型培养物保藏中心,保藏日期为2023年04月28日,保藏编号为CCTCC M 2023667,该菌筛选自自然发酵酸面团,该菌株分类命名为Lactiplantibacillus plantarum G8;本发明的植物乳植杆菌应用于黑麦麸皮‑小麦复合面包,能够显著改善黑麦麸皮‑小麦复合面包的硬度、胶着性、咀嚼性和弹性,能够改善烘焙损失率,还能够明显延长面面包的防霉效期至12天。
Description
技术领域
本发明涉及微生物领域及食品领域,尤其涉及一株植物乳植杆菌及其应用。
背景技术
黑麦麸皮中富含阿魏酸、膳食纤维等多种营养成分。随着消费者对健康的重视,富含膳食纤维的粗粮面包受到消费者的追捧。虽然粗粮面包营养健康,但是其保质期短并且口感较差。
具体而言,目前自然发酵的含有黑麦麸皮的小麦面包,保质期一般不超过9天,这给厂家和销售商销售、消费者食用都带来不便。另外,这类含黑麦麸皮的面包质构都较差,包括硬度、胶着性、咀嚼型和弹性等方面。
因此,采用一定的技术手段以延长含黑麦麸皮面包的保质期、提升其品质对于整个食品行业都具有重要意义。
发明内容
本发明的目的之一,是提供一株植物乳植杆菌,以解决上述问题。
为了实现上述目的,本发明采用的技术方案是这样的:一株植物乳植杆菌,,所述植物乳植杆菌的分类命名为Lactiplantibacillus plantarumG8,该菌株保藏于中国典型培养物保藏中心,保藏日期为2023年04月28日,保藏编号为CCTCC M 2023667。
本发明的目的之二,在于提供上述的植物乳植杆菌在制作面包中的应用。
作为优选的技术方案:所述面包为黑麦麸皮-小麦复合面包。
作为进一步优选的技术方案:所述黑麦麸皮-小麦复合面包的制作方法为:
(1)黑麦麸皮酸面团的制备:
称取纯净水与所述Lactiplantibacillus plantarumG8菌体混合,然后加入黑麦麸皮内并将其置于25~35℃恒温恒湿培养箱内发酵12~36 h,酸面团的初始接种量为108CFU/g;
(2)黑麦麸皮酸面团面包的制备:
将小麦粉、水、糖、盐、酵母及步骤(1)所得的黑麦麸皮酸面团,黄油除外,放入厨师机内,混合均匀,再加入黄油继续搅拌至面筋成型后取出;然后将面包面团置于25~35℃下进行一次发酵;一次发酵结束后将面团搓圆整型,25~35℃下二次发酵,在上/下火温度为160~180℃的烤箱内烘烤,即得。
作为更进一步优选的技术方案:步骤(2)中,所述小麦粉与黑麦麸皮酸面团的质量比为4:1。
与现有技术相比,本发明的优点在于:本申请的发明人筛选得到一株具有抗霉菌活性的植物乳植杆菌,并证明将其作为面包尤其是含黑麦麸皮面包的发酵剂,不仅能延长面包的保质期,而且能改善面包的加工、营养特性;采用本发明的菌株,能够将含黑麦麸皮面包的防霉效期延长至12天,这大大提高了生产商、销售商的销售便利性,提高经济效益,也易于消费者保存食用。
附图说明
图1为实施例1筛选得到的Lactiplantibacillus plantarumG8革兰氏染色图。
具体实施方式
下面将结合附图对本发明作进一步说明。
实施例1
菌株筛选:
取四川省雅安市三雅路菜市场自然发酵酸面团(常温放置约5 d)10 g,加入至90mL生理盐水混匀,经10倍梯度稀释,选择合适稀释度涂布于含0.5%碳酸钙的MRS固体培养基,37℃培养箱中培养48 h;挑选菌落为圆形、表面光滑、乳白、凸起、有溶钙圈的疑似乳酸菌菌落;经5次平板划线,得到纯化的单菌落,对其进行革兰氏染色和过氧化氢酶实验,选择革兰氏阳性、过氧化氢酶阴性菌株于20vol%甘油管保藏。
将保存的菌株按2vol%比例接种至新鲜MRS肉汤,37℃培养24h后测定MRS肉汤pH,选择产酸量最大的菌株。经DNA提取试剂盒提取分离菌株的基因组,采用通用引物27F(5'-AGAGTTTGATCMTGGCTCAG-3')和1492R(5'-GGTTACCTTGTTACGACTT-3')扩增其16S rDNA,将PCR扩增产物送至生工生物工程(上海)股份有限公司测序。将测序结果通过NCBI BLAST分析工具(http://www.ncbi.nlm.nih.gov/blast)比对,经分析鉴定得到植物乳植杆菌(Lactiplantibacillus plantarumG8),
其16S rDNA序列如下SEQ ID NO:1所示:
GTCAAATGCGGCGTCCTATAATGCAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCTAATGGTGACAGAGGG;
其革兰氏染色图见图1,并将其保藏于中国典型培养物保藏中心,保藏日期为2023年04月28日,保藏编号为CCTCC M 2023667。
实施例2
黑麦麸皮-小麦复合面包的制作,包括下述步骤:
(1)黑麦麸皮酸面团的制备:
将实施例1筛选得到的Lactiplantibacillus plantarumG8在MRS肉汤中培养至对数生长期,5000 g离心10 min收集菌体,用无菌生理盐水连续洗涤菌体两次,收集菌体。称取适量纯净水与菌体混合,加入黑麦麸皮内(酸面团的初始接种量为108CFU/g),DY=250,按公式(1)计算DY值,并将其置于30℃恒温恒湿培养箱内发酵24 h(后称“实施例”);
DY=(1)
式中:m粉表示面粉的质量(g),m水表示水的质量(g);
将按前述实施例1中同样筛菌方法筛选自三雅路菜市场、新鲜酸面团的同种乳酸菌另一株菌株Lactiplantibacillus plantarumG12(产酸能力中等)接种至黑麦麸皮制成的酸面团(后称“对照A”),未接种任何菌株的自然发酵黑麦麸皮酸面团作为对照组(后称“对照B”);
(2)黑麦麸皮酸面团面包的制备:
将80 g小麦粉、40 g水、6 g糖、1 g盐、1.5 g酵母及20 g黑麦麸皮酸面团(黄油除外)放入厨师机内,混合均匀,加入4 g黄油继续搅拌至面筋成型后取出。首先将面包面团置于30℃下发酵1 h;一次发酵结束后将面团搓圆整型,30℃下发酵1.5 h;在上/下火温度为170℃的烤箱内烘烤30 min。
对照A、对照B的黑麦麸皮酸面团面包的制作方法与上述步骤(2)完全相同,仅加入的酸面团不同;
得到三种黑麦麸皮-小麦复合面包,根据相应所采用的酸面团,分别为“实施例”、“对照A”和“对照B”。
实施例3
黑麦麸皮酸面团面包的质构测试
测试方法,参考Characteristics of sourdough bread fermented withPediococcus pentosaceus and Saccharomyces cerevisiae and its bio-preservativeeffect against Aspergillus flavus。
测试结果如下表1
表1不同面包的硬度、胶着性、咀嚼性及弹性
从表1可以看出,接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包(实施例)的硬度、胶着性及咀嚼性均最小,与两个对照组相比均有显著性差异。同时,接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包的弹性值最大,整体而言,Lactiplantibacillus plantarumG8能够改善面包的质构特性,获得更柔软的面包屑结构。
实施例4
黑麦麸皮酸面团面包的烘焙损失率测试
测试方法:对烘烤前的面包面团及烘烤后的面包进行称重,按如下公式计算各组面包的烘焙损失率。
烘焙损失率(%)=;
结果见下表2:
表2不同面包的烘焙损失率
从表2可以看出,接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包(实施例)的烘焙损失率最小,与两个对照组相比均有显著性差异。
实施例5
黑麦麸皮酸面团面包的防霉效期测试
测试方法:将冷却至室温的新鲜黑麦麸皮酸面团面包样品切成长宽高分别为100mm、100 mm、15 mm的切片,置于无菌采样袋内,并在室温下存储。
每天监测各组面包片表面的霉菌菌落,以评估面包的防霉效期。结果如表3所示,
表3不同面包的防霉效期
从表3可以看出,接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包(实施例)的防霉效期最长,与两个对照组相比均有显著性差异,实施例的防霉效期延长至12 d,相较于自然发酵面包,延长了33.3%。
实施例6
黑麦麸皮酸面团面包的感官评价
评价方法:感官评价小组由20名经培训的成员组成。将不同面包样品放入相同类型的塑料盒内,随机呈现给感官评价小组成员,过程中提供纯净水漱口。采用1~9分的享乐量表(9:非常喜欢,1:非常不喜欢)评估面包的颜色、味道、气味、质地以及整体可接受度;结果见表4;
表4不同面包的感官评价
从表4可以看出,接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包(实施例)在气味及质地方面的感官评分与对照组有显著差异,得分最高。接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包的整体可接受度高于对照B,但比对照A稍低。
实施例7
黑麦麸皮酸面团面包的体外蛋白消化率测试
测试方法:
(1)体外消化
参考Minekus等的方法(A standardised static in vitro digestion methodsuitable for food-an international consensus),并稍作修改。具体操作如下,首先将2g面包与10 mL SSF,0.5 mL含1500 U/mL α-淀粉酶的SSF,25 μL 0.3 mol/L CaCl2及975 μL超纯水充分混合,37℃下反应2 min。将10 mL混合物与7.5 mL SGF及5μL 0.3 M CaCl2充分混合,使用1 mol/L HCl溶液将pH调整至3,并添加1.6 mL含25000 U/mL猪胃蛋白酶的SGF,将混合物置于37℃下反应2 h。反应结束后,将20 mL胃食糜,11 mLSIF及40 μL 0.3mol/L CaCl2充分混合,使用1 mol/L NaOH溶液将溶液pH调节至7,添加5 mL含800 U/mL猪胰蛋白酶的SIF及0.4 g胆汁盐(食糜中约10 mmol/L),37℃下反应2 h。一式三份。模拟消化结束后,在95℃下加热5 min以终止反应。
(2)蛋白质消化率
将胃和肠相反应溶液分别与三倍体积的乙醇混合,在4℃下放置12 h以沉淀蛋白质(参考参考 Real meat and plant-based meat analogues have different in vitroprotein digestibility properties)。将样品以10000 g,4℃下离心20 min,将分离出的沉淀物悬浮于5 mL蛋白质提取缓冲液中,并将混合物以4000 g,4℃离心5 min。通过BCA蛋白质测定试剂盒测定上清液中蛋白质的含量。按公式(8)计算各组面包的蛋白质消化率。
蛋白质消化率(%)=(8)
式中W1是未消化蛋白质的含量(g);W0是消化前样品的蛋白质含量(g);
结果见表5:
表5不同面包的体外蛋白消化率(%)
从表5可以看出,接种Lactiplantibacillus plantarumG8黑麦麸皮酸面团面包(实施例)的体外蛋白消化率最高,与两个对照组均有显著差异。
以上所述仅是本发明的优选实施方式,应当理解本发明并非局限于本文所披露的形式,不应看作是对其他实施例的排除,而可用于各种其他组合、修改和环境,并能够在本文所述构想范围内,通过上述教导或相关领域的技术或知识进行改动。而本领域人员所进行的改动和变化不脱离本发明的精神和范围,则都应在本发明所附权利要求的保护范围内。
Claims (5)
1.一株植物乳植杆菌,其特征在于,所述植物乳植杆菌的分类命名为Lactiplantibacillus plantarum G8,该菌株保藏于中国典型培养物保藏中心,保藏日期为2023年04月28日,保藏编号为CCTCC M 2023667。
2.权利要求1所述的植物乳植杆菌在制作面包中的应用。
3.根据权利要求2所述的应用,其特征在于:所述面包为黑麦麸皮-小麦复合面包。
4.根据权利要求3所述的应用,其特征在于,所述黑麦麸皮-小麦复合面包的制作方法为:
(1)黑麦麸皮酸面团的制备:
称取纯净水与所述Lactiplantibacillus plantarum G8菌体混合,然后加入黑麦麸皮内并将其置于25~35℃恒温恒湿培养箱内发酵12~36 h,酸面团的初始接种量为108CFU/g;
(2)黑麦麸皮酸面团面包的制备:
将小麦粉、水、糖、盐、酵母及步骤(1)所得的黑麦麸皮酸面团,黄油除外,放入厨师机内,混合均匀,再加入黄油继续搅拌至面筋成型后取出;然后将面包面团置于25~35℃下进行一次发酵;一次发酵结束后将面团搓圆整型,25~35℃下二次发酵,在上/下火温度为160~180℃的烤箱内烘烤,即得。
5.根据权利要求4所述的应用,其特征在于,步骤(2)中,所述小麦粉与黑麦麸皮酸面团的质量比为4:1。
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