CN116478863A - 副干酪乳酪杆菌yys-k1及其应用 - Google Patents
副干酪乳酪杆菌yys-k1及其应用 Download PDFInfo
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Abstract
本发明属于微生物技术领域,具体涉及一种副干酪乳酪杆菌YYS‑K1及其应用,副干酪乳酪杆菌(Lacticaseibacillus paracasei)YYS‑K1于中国微生物菌种保藏管理委员会普通微生物中心的保藏编号为CGMCC No.26405,可应用于制备益生菌产品,具有改善肠道菌群结构、解酒、减肥等保健功能。
Description
技术领域
本发明属于微生物技术领域,具体涉及一种副干酪乳酪杆菌YYS-K1及其应用。
背景技术
酒在我国具有几千年的历史,其风味口感独特,是一种非常具有魅力的传统饮品。随着人类文明的不断发展,酒文化也得到了丰富和发展,常见的有啤酒、白酒、果酒、奶酒、茶酒等,具有巨大的社会效益和经济效益。适量饮酒具有提高食欲,帮助肠道消化,缓解疲劳忧虑,促进人体新陈代谢等功能。但若长期大量摄入,不仅对身体无益,还会造成许多疾病,例如酒精性肝炎、肝硬化、酒精性脂肪肝等。同时,也会对大脑产生影响,最终导致大脑反应迟钝,记忆力下降等不同程度的脑损伤。另外,过量饮酒还会强烈刺激人体的消化系统,造成食欲减退、营养不良等疾病,又或是引发其它器官的病变,严重时甚至会呼吸肌麻痹而导致猝死,对人体健康构成巨大威胁。
乙醇,也叫酒精,是各类酒饮品中的主要成分。乙醇在进入人体后80%以上是由肝脏代谢,而酒精性肝损伤的主要形成原因就是肝脏代谢产物造成的肝细胞紊乱,在嗜酒人群中具有较高的发病率。随着科技和社会的发展进步,以及人类对健康意识的逐渐加强,人们对过量饮酒的处理越来越重视,因此,对乙醇降解机理的探索成为了科学研究的热点。
我国具有非常丰富的中药资源,科研人员利用中药作为研究对象研制出了大量解酒产品,具有一定疗效。目前市售的大部分解酒产品主要都是以植物为原料制得,在服用后能够影响乙醇在人体内的代谢或者抑制人体对乙醇的吸收,从而达到醒酒的作用,但实质上并没有直接降解乙醇。部分产品虽具有一定的功效,但有可能会对人体产生毒副作用,不利于人体健康。并且市面上许多醒酒产品并没有实质的解酒作用,只是能缓解头痛,解酒效果并不理想。因此,研究一种高效、没有毒副作用且价格便宜的生物解酒药是非常必要的。
碳水化合物是微生物生长过程所必需的,如果微生物能将乙醇作为生长必需的碳源之一,就能达到降解乙醇的效果,但目前关于这方面的研究报道还比较少。
文献“一株乙醇降解菌株的筛选、鉴定及其培养条件的优化[J].酿酒科技,2011(06):17-20”从酸败的酒糟中筛选出一株降解乙醇能力相对较高的菌株,经鉴定为巴氏醋杆菌(Acetobacter pasteurianus)Ⅶ,其乙醇降解能力为0.72mL/d。
文献“乙醇降解菌种的筛选及其发酵乳产品解酒功效评价[J].食品科学,2020,41(02):107-113”从10株发酵性能良好的乳酸菌中筛选出一株乙醇降解效果较好的菌种,经鉴定为嗜酸乳杆菌(Lactobacillus acidophilus),培养24h后对乙醇降解率约为50%。
论文“快速解酒厌氧微生物的筛选鉴定及其解酒能力研究[D].浙江大学,2016”在韩国泡菜中筛得一株菌,该菌在20%乙醇浓度培养条件培养1h后对乙醇的降解率为21.1%,经测序鉴定该菌为海氏肠球菌(Enterococcus hirae)。
申请号为201911202646.7的中国发明专利公开了一种解酒护肝组合物及产品,包含植物提取物和益生菌,益生菌包含嗜酸乳杆菌NCFM、副干酪乳杆菌Lpc-37、乳双歧杆菌Bi-07、乳双歧杆菌Bl-04和乳双歧杆菌HN019,但其属于组合产品,其中的植物成分有大量文献证明具有解酒功效,不能证明其中的益生菌起到了解酒作用,且并未说明其使用的菌种任一单独使用能起到良好的解酒作用。
肥胖是当今社会影响人类健康的重要原因之一,大多都是由于食物摄取过量导致脂肪堆积,从而体重增大,在人群中普遍存在,因此,对减肥产品的开发也是当下研究的热点。现有的减肥药物大多是作用于神经中枢,对人体有较强的副作用,而另一类减肥产品——脂肪酶抑制剂,可以不经人体神经系统或血管发挥作用,对人体健康没有大的安全隐患,是一种安全的新型减肥制剂。胰脂肪酶由胰腺分泌,是作用于脂肪消化过程的关键酶,而胰脂肪酶抑制剂可以抑制肠道中的胰脂肪酶对脂肪进行分解,减少小肠粘膜对脂肪酸的吸收,从而达到减肥效果。微生物是多种酶制剂的重要来源,微生物代谢产物已广泛应用于食品行业,具有很高的应用价值。
文献“脂肪酶抑制剂产生菌筛选[J].中国抗生素杂志,2002(11):641-643”从土壤样品里分离筛选出8株具有脂肪酶抑制作用的菌株,其中抑制率最高的菌株LF高达80.6%,经鉴定为芽孢杆菌属,不属于可食用益生菌种类。
发明内容
为了克服现有技术的缺陷,本发明提供一种副干酪乳酪杆菌及其应用,本发明的副干酪乳酪杆菌,在高效降解乙醇的同时对胰脂肪酶有较高的抑制效果,可应用于解酒或减肥产品的开发,具有重要的应用价值。
为了解决上述技术问题,本发明采用的技术方案为:副干酪乳酪杆菌YYS-K1,分类命名为:副干酪乳酪杆菌YYS-K1,拉丁文学名:Lacticaseibacillus paracasei,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号:CGMCC No.26405。保藏地址:北京市朝阳区北辰西路1号院3号,保藏日期:2023年1月6日。
本发明还提供上述的副干酪乳酪杆菌YYS-K1在制备益生菌产品中的应用。
本发明还提供上述的副干酪乳酪杆菌YYS-K1在制备耐酸和/或耐胆盐的食品或保健品中的应用。
本发明还提供上述的副干酪乳酪杆菌YYS-K1在制备解酒剂中的应用。
本发明还提供上述的副干酪乳酪杆菌YYS-K1在制备减肥产品中的应用。
本发明还提供一种包含上述副干酪乳酪杆菌YYS-K1的菌剂。
本发明的有益效果在于:本发明提供了一种副干酪乳酪杆菌YYS-K1,具有高乙醇降解能力和胰脂肪酶抑制率,在MRS培养基中添加20%的乙醇,其乙醇降解率为43.9%,在MRS培养基中添加40%的乙醇,其乙醇降解率为48.6%;其发酵液对胰脂肪酶抑制率为64.9%,同时YYS-K1具有良好的耐酸、耐胆盐性,其所表现的高乙醇降解能力、胰脂肪酶抑制能力和耐酸耐胆盐性使其能够应用于制备改善肠道菌群结构的益生菌产品,以及解酒或减肥产品,且不存在植物原料由于成分复杂易引起毒副作用的问题,为解酒或减肥产品的开发提供了新的方向和更多的可能。
附图说明
图1所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1分离过程中的溶钙圈;
图2所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1的菌落形态;
图3所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1在不同乙醇浓度下的活菌数;
图4所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1降解乙醇试验中乙醇标准曲线;
图5所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1抑制胰脂肪酶试验中4-硝基苯酚(PNP)标准曲线;
图6所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1的革兰氏染色结果;
图7所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1的16S rRNA扩增产物琼脂糖凝胶电泳图;
图8所示为本发明具体实施方式的副干酪乳酪杆菌YYS-K1基于16S rRNA序列的系统发育树。
具体实施方式
为详细说明本发明的技术内容、所实现目的及效果,以下结合实施方式并配合附图予以说明。
本发明提供一种副干酪乳酪杆菌(Lacticaseibacillus paracasei)YYS-K1,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号:CGMCC No.26405,保藏地址:北京市朝阳区北辰西路1号院3号,保藏日期:2023年1月6日。
实施例1:副干酪乳酪杆菌YYS-K1的分离
从农家自制杨梅酒中无菌采样,采用涂布平板法,取5g杨梅酒样品放入无菌均质袋中,做好标记,加入45mL 0.85%的生理盐水后完全拍打混匀,得到样品。然后吸取100μL样品进行10倍系列的梯度稀释,分别吸取稀释倍数为10-4、10-5、10-6的样品100μL,涂布于含2.5% CaCO3的MRS平板上,37℃倒置培养24h。挑取长势良好,溶钙圈大的菌落(参见图1),通过平板划线分离法反复分离纯化,直至得到单菌落(参见图2),将该分离菌株命名为YYS-K1,编号,加入甘油于-80℃菌库保藏。
实施例2:副干酪乳酪杆菌YYS-K1降解乙醇试验
2.1乙醇耐受能力分析
将副干酪乳酪杆菌YYS-K1菌种活化至二代,按3%的接种量接种至添加量为0%、10%、20%、30%、40%、50%无水乙醇的MRS培养基中,每组三个平行,37℃培养24h,培养结束后采用平板倾注法测定活菌数,分析其乙醇耐受能力,结果参见图3所示。
2.2溶液配制
乙醇标准液:精密吸取无水乙醇,加入蒸馏水中,配制成体积分数分别为0%、10%、30%、50%、70%、90%的乙醇标准液。
重铬酸钾-浓硫酸溶液:称取重铬酸钾1g溶于25mL蒸馏水中,边搅拌边缓慢加入4mL浓硫酸(95%~98%%)。
2.3乙醇标准曲线绘制
向试管中分别加入0%、10%、30%、50%、70%、90%的乙醇标准液1.0mL,每个试管中加入重铬酸钾-浓硫酸溶液3.0mL,充分摇匀,盖好盖子;将试管置于沸水浴中10min,使重铬酸钾-浓硫酸溶液充分氧化;取出试管,于波长610nm处测定重铬酸钾-浓硫酸溶液的吸光值。以乙醇浓度为横坐标,吸光值为纵坐标,绘制标准曲线(参见图4)。
2.4待测样品的检测分析
分别配置乙醇体积分数为20%、40%的乙醇-MRS液体培养基,取5mL液体培养基于试管中,加入活化至二代的YYS-K1菌液200μL,混匀后放于37℃摇床振荡培养24h。以不加菌液的乙醇-MRS液体培养基为对照,利用重铬酸钾-浓硫酸法测定乙醇残留量,分析菌种的乙醇降解能力。每个样品做3个平行,610nm测定吸光值。
由测得的标准曲线获得乙醇浓度与OD610 nm吸光值之间的线性关系,乙醇降解率=(乙醇原始浓度-乙醇剩余浓度)/乙醇原始浓度×100%,样品检测得出YYS-K1在乙醇浓度为20%时,其乙醇降解率可达到43.9%,在乙醇浓度为40%时,其乙醇降解率可达到48.6%。
MRS液体培养基:葡萄糖20.0g,胰蛋白胨10.0g,牛肉膏10.0g,酵母浸粉5.0g,吐温80 1.0mL,磷酸氢二钾2.0g,柠檬酸铵2.0g,无水乙酸钠5.0g,硫酸镁0.5g,一水硫酸锰0.25g,去离子水1L,pH 6.5(加1.5%琼脂为MRS固体培养基)。
本发明提供的副干酪乳酪杆菌YYS-K1具有良好的降解乙醇能力,可应用于制备解酒剂。
实施例3:副干酪乳酪杆菌YYS-K1胰脂肪酶抑制试验
3.1溶液配制
Tris-HCl缓冲液:准确称取6.05g Tris,加入1L蒸馏水溶解,再加入0.1%阿拉伯树胶粉和0.2%脱氧胆酸钠,调节pH=8.0。
胰脂肪酶溶液(PPL):称取240mg胰脂肪酶,加入200mL Tris-HCl缓冲液溶解,混匀静置,6000r/min离心5min,取上清液,放于-20℃保存。
底物溶液:称取20mg 4-硝基苯棕榈酸酯(PNPP)溶于5mL异丙醇,再用Tris-HCl缓冲液定容至100mL。
3.2PNP标准曲线的绘制
准确称取4-硝基苯酚(PNP)标准品7.0mg,溶于二甲基亚砜(DMSO),定容至10mL,制成浓度为5mmol/L母液。使用Tris-HCl缓冲液将母液PNP稀释至0.05mmol/L、0.25mmol/L、0.1mmol/L、0.2mmol/L、0.3mmol/L,测定不同浓度的PNP在405nm下的吸光值。以PNP的浓度为横坐标,吸光值为纵坐标,绘制标准曲线(参见图5)。
3.3样品的检测分析
将副干酪乳酪杆菌YYS-K1活化至二代,8000r/min离心10min,上清液即为待测样品溶液。试验分为A、B、C、D四组,各反应物剂量见表1,使用标准96孔板,每组5个平行。实验条件模拟人体环境进行设计,依次加入pH=8.0的Tris-HCl缓冲液、待测样品溶液和胰脂肪酶溶液,混合均匀,于37℃水浴保温10min,结束后,取出96孔板,加入底物溶液(PNPP),充分混匀,于37℃水浴反应20min。PNPP在胰脂肪酶的作用下能水解产生PNP,PNP在405nm处有最大吸光值,测定其吸光值,计算副干酪乳酪杆菌YYS-K1对胰脂肪酶的抑制率,计算公式如下:
抑制率=[1-(C-D/A-B)]×100%
其中A为对照组吸光值,B为对照空白组吸光值,C为样品组吸光值,D为样品空白组吸光值。
由测得的标准曲线可知PNP与OD405nm吸光值之间的线性关系,样品检测得出YYS-K1对胰脂肪酶PPL的抑制率可达64.9%。
本发明提供的副干酪乳酪杆菌YYS-K1具有良好的胰脂肪酶抑制效果,可应用于减肥产品开发。
表1
实验组 | Tris-HCl(μL) | PPL(μL) | PNPP(μL) | 待测样品(μL) |
A | 50 | 50 | 100 | 0 |
B | 100 | 0 | 100 | 0 |
C | 30 | 50 | 100 | 20 |
D | 80 | 0 | 100 | 20 |
实施例4:副干酪乳酪杆菌YYS-K1的鉴定
4.1生理生化试验
将筛选纯化的菌株YYS-K1进行革兰氏染色(参见图6)及过氧化氢酶试验,并对其生理生化指标进行测定,试验结果对照《伯杰氏系统细菌学手册第八版》进行菌种的初步判定。试验得出该筛选菌株YYS-K1革兰氏染色为紫色,呈阳性。其细胞形状为杆状,接触酶和氧化酶为阴性,无芽孢形成。
4.2 16S rRNA鉴定
根据细菌基因DNA提取试剂盒的说明提取YYS-K1该未知菌株DNA,以DNA为模板进行16S rRNA基因的PCR扩增。
扩增引物使用通用引物:
27F:5′-AGAGTTTGATCCTGGCTCAG-3′(SEQ ID NO.1)。
1492R:5′-TACGGCTACCTTGTTACGACTT-3′(SEQ ID NO.2)。
PCR反应体系:DNA 2μL,27F 2μL,1492R 2μL,Premix Ex Taq 25μL,ddH2O 19μL。
PCR反应条件:94℃预变性3min;94℃变性30s,55℃退火30s,72℃延伸1min,30次循环;最后72℃延伸5min。
YYS-K1提取DNA基因组后16S rRNA扩增片段参见图7所示。然后将PCR扩增产物送去DNA测序(广州擎科生物工程有限公司)。测序序列结果在NCBI数据库中用Blast软件搜索近似序列,将所测得的序列和从基因库中获得的相关种属的16S rRNA基因序列进行比对,运用Mega7.0软件构建系统发育树,结果参见图8所示。
16S rRNA基因序列测定结果如下:
GCAGTCGAACGAGTTCTCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAGGTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTTTTAGGGAGCGAGCCGTCTAA(SEQ ID NO.3)。
根据该菌株的细胞形态、生理生化特征、16S rRNA基因序列等数据综合分析,参考《伯杰氏系统细菌学手册第八版》,鉴定该菌株为副干酪乳酪杆菌(Lacticaseibacillusparacasei)。
实施例5:副干酪乳酪杆菌YYS-K1耐酸性和耐胆盐试验
5.1模拟胃液耐酸性试验
称取0.2g NaCl与0.35g胃蛋白酶溶解于适量蒸馏水中,共配制3份,用1.0mol/L盐酸分别调节pH值为1.5、2.5、3.5,用100mL容量瓶定容至刻度。将副干酪乳酪杆菌YYS-K1活化后培养24h的发酵液按2%的接种量分别接入不同pH值的模拟人工胃酸溶液中,37℃水浴恒温处理0、2、4h后,分别10倍梯度稀释,振荡均匀后取100μL菌液倒板,37℃下培养24h,计算YYS-K1活菌数,结果见表2所示。
表2
从表1可以看出在不同pH的模拟人工胃液中,副干酪乳酪杆菌YYS-K1活菌数不同,随着pH值的升高,活菌数在不断增多,存活率也在不断提高。而在相同pH的模拟人工胃液中,随着处理时间的增加,活菌数减少。但在不同pH的模拟人工胃液中,处理2h后活菌数均保持较高水平,表明YYS-K1在经过人体胃液消化后仍能有较高存活数量,对胃酸的耐受性优良。
5.2模拟肠液耐胆盐试验
将副干酪乳酪杆菌YYS-K1活化后培养24h的发酵液按2%接种量,分别接入质量浓度为0.03g/mL、0.3g/mL和0.5g/mL的猪胆盐溶液中,37℃水浴恒温处理0、2、4h后,分别10倍梯度稀释,振荡均匀后取100μL菌液倒板,37℃下培养24h后,计算YYS-K1活菌数,结果见表3所示。
表3
从表2可以看出副干酪乳酪杆菌YYS-K1在模拟人工肠液胆盐不同起始质量浓度下,其活菌数不同,存活率也不同。随着模拟人工肠液胆盐质量浓度的增加,起始活菌数在逐渐减少,处理时间越长,存活率也逐渐下降。但在最接近人体的胆盐浓度(人体小肠中的胆盐浓度一般为0.03~0.3g/mL)处理2h后,仍具有较高存活数量,表明YYS-K1在人体内经胃肠消化后仍能有较多活菌,以进行乙醇代谢。
本发明的副干酪乳酪杆菌YYS-K1具有良好的耐胃酸、耐胆盐作用,可以改善肠道菌群结构,用于制备耐酸和/或耐胆盐的食品或保健品,具有较高的商业应用价值。
实施例6:
一种菌剂(益生菌产品),包括副干酪乳酪杆菌YYS-K1和保护剂;保护剂由100g/L脱脂奶粉、30mL/L甘油、150g/L海藻糖与10g/L L-谷氨酸钠组成。
上述菌剂的制备方法为:
S1、将副干酪乳酪杆菌YYS-K1按照培养基质量的3%的接种量接种于121℃灭菌15min的培养基中,然后在温度37℃下培养20h后,于4℃下6000r/min离心20min,弃上清,加入pH=7.2磷酸盐缓冲液清洗2~4次,得到菌泥,再用保护剂重悬,得到菌液浓度为1010CFU/mL的悬浮液;
其中,培养基由培养基总质量0.5%葡萄糖、1.5%胰蛋白胨、0.3%酵母浸膏和余量水组成,pH=6.8;
S2、将S1的悬浮液在温度37℃下预培养60min,再进行冷冻干燥,得到含有大于1×1010CFU/g的活性副干酪乳酪杆菌YYS-K1的菌剂。
实施例7:
一种益生菌产品,包括副干酪乳酪杆菌YYS-K1与其他配料。益生菌产品中,副干酪乳酪杆菌YYS-K1的活菌数不低于1×106CFU/mL或1×106CFU/g。所述配料包含益生元、填充剂、酸味剂、溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、润滑剂、润湿剂、渗透压调节剂、稳定剂、助流剂、矫味剂、防腐剂、包衣材料、芳香剂、抗黏合剂、整合剂、渗透促进剂、pH值调节剂、缓冲剂、增塑剂、表面活性剂、消泡剂、增稠剂、包合剂、保湿剂、吸收剂、稀释剂、絮凝剂与反絮凝剂以及助滤剂中的一种或几种。
益生菌产品的剂型可以为固体饮料、液体饮料、压片糖果、颗粒剂、胶囊剂、片剂、丸剂或口服液。
本实施例中,提供一种剂型为固体饮料的益生菌产品(副干酪乳酪杆菌YYS-K1固体饮料),按重量份计,包括活性菌粉10份、木糖醇24.8份、全脂奶粉24份、发酵桑葚粉12份、麦芽糊精23份、低聚半乳糖5.6份、低聚果糖0.1份和无水柠檬酸0.5份。
该固体饮料的制备方法为:
S1、将所述副干酪乳酪杆菌YYS-K1菌种用液体培养液培养,收集和洗涤菌体,添加辅料(实施例6的保护剂),干燥制备成活性菌粉;
S2、将活性菌粉、木糖醇、全脂奶粉、发酵桑葚粉、麦芽糊精、低聚半乳糖、低聚果糖和无水柠檬酸混合均匀后得到副干酪乳酪杆菌YYS-K1活菌数不低于1×108CFU/g的固体饮料。
所有实施例中未明确指出的实验操作方法为本领域常规的实验操作方法,涉及的试剂或仪器均可从正规渠道商购获得。
综上所述,关于副干酪乳酪杆菌的解酒功能性或胰脂肪酶抑制性菌株,尚未见报道,本发明提供的副干酪乳酪杆菌YYS-K1,与巴氏醋杆菌、嗜酸乳杆菌同属不同种;与海氏肠球菌不同种属,其不同于现有益生菌解酒制剂,需要多种搭配使用,单独使用即可达到良好的降解乙醇的能力,在MRS培养基中添加20%的乙醇,其乙醇降解率为43.9%,在MRS培养基中添加40%的乙醇,其乙醇降解率为48.6%。同时副干酪乳酪杆菌YYS-K1具有较高的胰脂肪酶抑制活性和良好的耐酸、耐胆盐性,不但能够应用于制备改善肠道菌群结构的益生菌产品,将其应用于解酒产品或减肥产品时,也能发挥更长久和更稳定的作用;同时副干酪乳酪杆菌YYS-K1作为单一成分,不存在植物原料由于成分复杂或者复配后组分间作用不明,易引起毒副作用的问题,为解酒或减肥产品的开发提供了新的方向和更多的可能。
以上所述仅为本发明的实施例,并非因此限制本发明的专利范围,凡是利用本发明说明书及附图内容所作的等同变换,或直接或间接运用在相关的技术领域,均同理包括在本发明的专利保护范围内。
Claims (8)
1.副干酪乳酪杆菌YYS-K1,其特征在于,分类命名为:副干酪乳酪杆菌YYS-K1,拉丁文学名:Lacticaseibacillus paracasei,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号:CGMCC No.26405。
2.权利要求1所述的副干酪乳酪杆菌YYS-K1在制备益生菌产品中的应用。
3.权利要求1所述的副干酪乳酪杆菌YYS-K1在制备耐酸和/或耐胆盐的食品或保健品中的应用。
4.权利要求1所述的副干酪乳酪杆菌YYS-K1在制备解酒剂中的应用。
5.权利要求1所述的副干酪乳酪杆菌YYS-K1在制备减肥产品中的应用。
6.一种菌剂,其特征在于,包括权利要求1所述的副干酪乳酪杆菌YYS-K1。
7.根据权利要求6所述的菌剂,其特征在于,所述副干酪乳酪杆菌YYS-K1的活菌数不低于1×106CFU/mL或1×106CFU/g。
8.根据权利要求6所述的菌剂,其特征在于,还包括益生元、填充剂、酸味剂、溶剂、增溶剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、润滑剂、润湿剂、稳定剂、助流剂、矫味剂、防腐剂、包衣材料、芳香剂、抗黏合剂、整合剂、增稠剂和包合剂中的至少一种。
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