CN113930367A - 一株具有降胆固醇性能的乳酸菌及其应用 - Google Patents
一株具有降胆固醇性能的乳酸菌及其应用 Download PDFInfo
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- CN113930367A CN113930367A CN202111342225.1A CN202111342225A CN113930367A CN 113930367 A CN113930367 A CN 113930367A CN 202111342225 A CN202111342225 A CN 202111342225A CN 113930367 A CN113930367 A CN 113930367A
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Abstract
本发明提供一株具有降胆固醇性能的乳酸菌及其应用。本发明提供的食窦魏斯氏菌菌株为食窦魏斯氏菌LMLC4560‑1菌株(Weissella cibaria LMLC4560‑1),其于2021年7月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCC NO:M 2021811。本发明提供的食窦魏斯氏菌LMLC4560‑1菌株具有高效的降胆固醇能力和较强的抗氧化能力,同时具有良好的耐胃肠道逆环境性能。
Description
技术领域
本发明属于微生物领域,具体涉及一株具有降胆固醇性能的乳酸菌及其应用。
背景技术
魏斯氏菌属于乳酸菌,分布广泛,食物、土壤、胃肠道和呼吸道等环境中均含有多种魏斯氏菌,其中发酵食品是魏斯氏菌的丰富来源。发酵食品来源的一些魏斯氏菌由于基因组中含有编码铜氧化酶的sufI基因,能显著降低发酵食品中的生物胺。魏斯氏菌代谢产生细菌素可预防致腐微生物的生长,延长食品保质期的作用。魏斯氏菌还具合成低聚糖的能力,其产物可作为调味剂用于糖尿病人、肥胖病人的食物中,也可用于化妆品和饲料行业。研究表明,食窦魏斯氏菌有抗癌、免疫调节、抑制炎症水平和抗氧化的活性。
人类的高胆固醇血症已被证明是引起动脉粥样硬化,导致心脑血管疾病的主要原因。高胆固醇与许多代谢综合症相关,包括肥胖、高血脂、高血糖和高血压等。当人体内血清中总胆固醇浓度超过7.28mmol/L,急性冠心病发病危险程度也大幅增加。因此,通过膳食遏制来降低胆固醇的摄入并开发具有调节机体胆固醇含量的食品是更为健康有效的途径,而具备降低胆固醇功效的益生菌是其中颇有发展前景的功能性成分。
目前降低胆固醇主要依靠药物治疗,他汀类药物在降低血清胆固醇的同时,但也造成了机体肌肉痉挛以及横纹肌溶解等不良反应。另一种主要方式则是通过膳食食疗,虽取得一定的效果,但是其效果随着时间的延长会大大减弱,同时,长期的食疗是很难坚持下来的。因此,功能性食品便应运而生,既没有副作用也不需要长期的坚持,人们对这类产品的关注越来越高,这不仅仅是其本身的营养价值高,也迎合了公众的健康需求和食品工业的发展要求。所以,研究和利用具有降胆固醇益生菌开发研制降胆固醇功能食品及微生态制剂,对改善人们的健康水平有重要的意义。
发明内容
本发明的目的是提供一株具有高效降胆固醇能力的食窦魏斯氏菌菌株。
本发明提供的食窦魏斯氏菌菌株还具有较强的抗氧化能力和良好耐胃肠道逆环境。
本发明提供的食窦魏斯氏菌菌株从西藏牦牛奶中分离获得,具有食品属性。
本发明提供一株食窦魏斯氏菌菌株,所述食窦魏斯氏菌菌株为食窦魏斯氏菌LMLC4560-1菌株(Weissella cibaria LMLC4560-1),所述食窦魏斯氏菌LMLC4560-1菌株(Weissella cibaria LMLC4560-1)于2021年7月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCC NO:M 2021811。
优选地,所述食窦魏斯氏菌LMLC4560-1菌株的16S rDNA基因序列如SEQ ID NO.2所示。
本发明还提供一种食窦魏斯氏菌菌剂的发酵制备方法,所述方法包含如下步骤:培养所述的食窦魏斯氏菌菌株。
优选地,所述制备方法包括如下步骤:
(1)将所述的食窦魏斯氏菌菌株放大培养;
(2)将步骤(1)得到的产物加入到液体培养基中,在26-32℃条件下发酵培养。
本发明还提供一种菌剂,所述菌剂通过所述的制备方法得到。
本发明还提供所述的食窦魏斯氏菌菌株或所述的菌剂发酵获得的发酵产物。
优选地,所述发酵产物具有降胆固醇性能;
优选地,所述发酵产物具有自由基清除能力。
优选地,所述发酵产物作用于胆固醇,胆固醇的降低率大于87%;
或者,优选地,所述发酵产物对DPPH·自由基清除率大于90%。
本发明还提供所述的发酵产物的制备方法,所述制备方法通过包括培养所述的食窦魏斯氏菌菌株或所述的菌剂制备得到。
优选地,所述制备方法包括如下步骤:
(1)将所述的食窦魏斯氏菌菌株或所述的菌剂放大培养;
(2)将步骤(1)得到的产物加入到液体培养基中,在26-32℃条件下发酵培养得到所述发酵产物。
本发明还提供所述的食窦魏斯氏菌菌株或所述的菌剂在降低胆固醇、降脂、减肥、抗氧化或抗衰老产品中的应用。
优选地,所述产品为食品、化妆品、保健品或药品。
本发明提供的食窦魏斯氏菌LMLC4560-1菌株属于乳酸菌,并且来源于西藏地区牧民家牦牛奶,具有食品属性。本发明提供的食窦魏斯氏菌LMLC4560-1菌株具有高效的降胆固醇能力和较强的抗氧化能力,同时具有良好的耐胃肠道逆环境性能,补充了降胆固醇功能的益生菌的菌种资源。采用本发明提供的食窦魏斯氏菌发酵食品可以丰富食品的营养成分,还具有改善食品的适口性、消化性等作用。食品原料经本发明提供的食窦魏斯氏菌发酵后,能使蛋白质、脂肪和糖类分解为人体更易吸收的预消化状态,同时还能增加可溶性钙、磷、铁和某些B族维生素的含量,提高消化吸收性能和营养价值。
菌种保藏信息
本发明提供的食窦魏斯氏菌菌株为食窦魏斯氏菌LMLC4560-1菌株(Weissellacibaria LMLC4560-1)于2021年7月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCC NO:M 2021811,保藏地址:中国.武汉.武汉大学,邮政编码:430072;电话:027-68754052。
本发明实施例中的植物乳杆菌LP-23(Lactobacillus plantarum LP-23)于2020年6月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCC NO:M 2020175,保藏地址:中国.武汉.武汉大学,邮政编码:430072;电话:027-68754052。
附图说明
图1所示为实施例3中所述胆固醇标准曲线;
图2所示为食窦魏斯氏菌LMLC4560-1菌株菌落形态图;
图3所示为食窦魏斯氏菌LMLC4560-1菌株形态特征图。
具体实施方式
本发明提供的食窦魏斯氏菌LMLC4560-1菌株从西藏山南市加查县拉绥乡牦牛奶中分离获得。本发明提供的食窦魏斯氏菌LMLC4560-1菌株是将西藏牦牛奶中富集得到的菌株经放线菌酮和硫酸粘菌素筛选得到的具有产酸性能的革兰氏阳性菌株。本发明提供的食窦魏斯氏菌LMLC4560-1菌株具有食品属性,且具有高效的降胆固醇能力和较强的抗氧化能力,同时具有良好的耐胃肠道逆环境性能。用于发酵食品,可以丰富食品的营养成分,改善食品的适口性、消化性。
实施例中所用试剂具体来源列于下面的表1中。
表1本发明所用试剂及仪器来源信息
试剂/仪器 | 纯度/型号 | 生产厂商 |
酵母浸粉 | 99% | 安琪酵母股份有限公司 |
葡萄糖 | 99% | 沂水大地药业有限公司 |
石蕊牛乳培养基 | BR | 北京索莱宝生物科技有限公司 |
MRS肉汤 | BR | 北京奥博星生物技术有限公司 |
BL琼脂培养基 | BR | 北京索莱宝生物科技有限公司 |
革兰氏染色液 | 99% | 北京索莱宝生物科技有限公司 |
Super PCR Mix | 99.8% | 北京擎科新业生物技术有限公司 |
放线菌酮 | >94% | 湖南合众生物科技有限公司 |
硫酸粘菌素 | BR | 湖南合众生物科技有限公司 |
胆固醇 | AR | Sigma-Aldrich |
邻苯二甲醛 | AR | 麦克林世试剂网 |
紫外分光光度计 | SP-754 | 上海光谱仪器有限公司 |
高速离心机 | Minispin | eppendorf |
PCR仪 | C1000Touch | Bio-rad |
生化培养箱 | SPX250BSH-II | 上海新苗医疗器械制造有限公司 |
实施例1革兰氏阳性杆菌的富集、分离和纯化
①菌株富集:取1ml西藏山南市加查县拉绥乡牦牛奶样品进行梯度稀释至10-3,取2ml稀释液加入到20ml石蕊牛乳培养基中,然后放到37℃培养箱培养,待培养基酸化凝固并呈现粉红色时取出。
②培养单菌落:将石蕊牛乳培养基菌液稀释浓度到10-5-10-6,接种到含有10ppm放线菌酮和10ppm硫酸粘菌素的BL琼脂培养基,在37℃培养24~48h,待菌落形成。
③筛选产酸菌种:将初筛得到的单菌落,在加0.2%CaCO3的BL琼脂培养基中划线培养,培养24h-48h,结束培养后观察有无透明圈。
④产酸菌种的纯化:选择透明圈大的单菌落,划线纯化。重复纯化3次。
⑤革兰氏染色初步鉴定:对纯化好的产酸菌株进行革兰氏染色,筛选菌体是紫色的革兰氏阳性杆菌进行后续研究。
实施例2制备革兰氏阳性杆菌的发酵液
①活化菌株:用移液枪吸取1ml甘油管保存的革兰氏阳性杆菌的菌液,接种到9mlMRS液体培养基中,37℃,活化培养24h。
②培养种子液:用移液枪吸取1ml革兰氏阳性杆菌活化培养液,接种到9mlMRS液体培养基中,37℃,培养24h。
③发酵:用移液枪吸取5ml革兰氏阳性杆菌种子培养液,接种到45mlMRS-CHOL(胆固醇MRS培养基)液体培养基中,37℃,培养48h。
④离心发酵液:将发酵48h的发酵液12000rpm离心10min,收集上清。即得到革兰氏阳性杆菌的发酵液上清。
实施例3革兰氏阳性杆菌降低胆固醇能力的测定
胆固醇标准曲线的绘制
取浓度为1mg/mL胆固醇储存液0,0.1,0.2,0.3,0.4,0.5mL分别于10mL干净试管中,60℃水浴,待乙醇挥发干净,加2mL现配的邻苯二甲醛显色剂,振荡均匀后,室温静置10min,将1mL浓硫酸缓慢加入,混匀,静置20min,560nm波长下比色。横坐标为胆固醇浓度,纵坐标为吸光值,作标准曲线。得到的标准曲线如图1所示。
测定实施例2制备的革兰氏阳性杆菌的发酵液上清降低胆固醇的能力,用邻苯二甲醛法测定发酵液胆固醇的含量。
以未接种革兰氏阳性杆菌的胆固醇培养基为空白对照组。然后按照如下公式计算革兰氏阳性杆菌的胆固醇降低率。结果如表2。
X=(1-A/B)*100%;
其中:X-革兰氏阳性杆菌的胆固醇降低率;
A-革兰氏阳性杆菌的发酵液上清中胆固醇含量;
B-空白对照组上清中胆固醇含量。
表2革兰氏阳性杆菌对胆固醇的降低率
菌株编号 | 胆固醇降低率 | 菌株编号 | 胆固醇降低率 |
LMLC4560-1 | 87.3% | LMLC4560-11 | 41.3% |
LMLC4560-2 | 18.0% | LMLC4560-12 | 39.0% |
LMLC4560-3 | 22.7% | LMLC4560-13 | 40.6% |
LMLC4560-4 | 20.1% | LMLC4560-14 | 20.6% |
LMLC4560-5 | 20.8% | LMLC4560-15 | 35.2% |
LMLC4560-6 | 5.6% | LMLC4560-16 | 30.5% |
LMLC4560-7 | 32.3% | LMLC4560-17 | 23.5% |
LMLC4560-8 | 29.6% | LMLC4560-18 | 35.2% |
LMLC4560-9 | 18.0% | LMLC4560-19 | 48.7% |
LMLC4560-10 | 22.7% | LP-23 | 36.5% |
如表2结果显示:牦牛奶中分离、筛选、纯化的19株革兰氏阳性杆菌和本实验室自有植物乳杆菌LP-23,共计20株菌株中降胆固醇能力比较强的10株分别是:LMLC4560-1、LMLC4560-7、LMLC4560-11、LMLC4560-12、LMLC4560-13、LMLC4560-15、LMLC4560-16、LMLC4560-18、LMLC4560-19、LP-23。
该LP-23即植物乳杆菌LP-23(Lactobacillus plantarum LP-23),于2020年6月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCC NO:M2020175。
该植物乳杆菌LP-23由如下方式分离纯化获得:
①菌株富集:取1ml中国湖北省宜昌市某牛奶厂牛奶样品进行梯度稀释至10-3,取2ml稀释液加入到20ml石蕊牛乳培养基中,然后放到37℃培养箱培养,待培养基酸化凝固并呈现粉红色时取出;
②培养单菌落:将石蕊牛乳培养基菌液稀释浓度到10-5-10-6,接种到含有10ppm放线菌酮和10ppm硫酸粘菌素的BL琼脂培养基,在37℃培养24~48h,待菌落形成;
③筛选产酸菌种:将初筛得到的单菌落,在加0.2%CaCO3的BL琼脂培养基中划线培养,培养24h-48h,结束培养后观察有无透明圈;
④产酸菌种的纯化:选择透明圈最大的单菌落,即植物乳杆菌LP-23菌株。
对植物乳杆菌LP-23菌株进行16SrDNA测序及菌种鉴定。采用的引物为:正向引物27F:5'-AGAGTTTGATCATGGCTCAG-3';反向引物1492R:5'-ACGGCTACCTTGTTACGACTT-3'。PCR反应体系是:25μlMix;1μl样品;20μl ddH2O;2μl 27F;2μl 1492R;PCR反应温度:98℃,2min;98℃,10s,58℃,10s,72℃,7s,35x;72℃,2min;4℃保存。对PCR产物剂型测序,测得的植物乳杆菌LP-23菌株16SrDNA基因序列SEQ ID NO.1如下所示:
实施例4革兰氏阳性杆菌的人工消化液的耐性测定
测定实施例3中筛选的降胆固醇能力较大的10株革兰氏阳性杆菌对人工消化液的耐性。
①人工消化液的配制
人工胃液:NaCl 0.20g/100mL,胃蛋白酶0.35g/mL,用1mol/L的HCl调整pH值为2.5后,过滤除菌备用。
人工肠液:将下述A液和B液以2:1混合即为人工肠液。A.胰腺液:重碳酸钠1.1g/100mL,NaCl 0.2g/100mL,胰蛋白酶0.1g/100mL,调整pH值为8.0,过滤除菌备用。B.胆汁液:胆汁酸盐1.8g/100mL,调整pH值为6.8,过滤除菌备用。
②人工消化液耐性测定
挑取革兰氏阳性杆菌的单菌落在MRS液体培养基中培养过夜后,取培养液1mL分别接种于9mL的人工胃液或人工肠液中,37℃培养2小时,测定其活菌数。接种于9mL MRS液体培养基的革兰氏阳性杆菌作为阳性对照。
存活率(%)=(人工消化液中活菌数量/对照中活菌数量)*100%
表3革兰氏阳性杆菌经人工胃液和人工肠液处理后存活率
如上表3中所示,革兰氏阳性杆菌LMLC4560-1在人工胃液和人工肠液中均具有较高的存活率,分别达到96.35%和92.36%。
实施例5革兰氏阳性杆菌发酵液上清对DPPH·自由基清除率的测定
测定实施例3中筛选的降胆固醇能力较大的10株革兰氏阳性杆菌发酵液上清对DPPH·自由基清除率。测定方法如下:
①活化菌株:用移液枪吸取1ml甘油管保存的革兰氏阳性杆菌菌液,接种到9mlMRS液体培养基中,37℃,活化培养24h。
②培养种子液:用移液枪吸取1ml活化培养液,接种到9mlMRS液体培养基中,37℃,培养24h。
③发酵:用移液枪吸取5ml种子培养液,接种到45mlMRS液体培养基中,37℃,培养24h。
④制备发酵液上清:将发酵24h的发酵液5000rpm离心4min,收集上清。即得到革兰氏阳性杆菌的发酵液上清。
⑤取3mL待测发酵液与等体积的0.16mmol/L的DPPH溶液混匀(A1管);取等体积的无水乙醇与0.16mmol/L的DPPH溶液混匀(A2管);取等体积的无水乙醇与待测液混匀(A3管);30℃避光反应30min后,以蒸馏水为空白组,在519nm下测A1、A2、A3管的吸光度值,分别记作ODA1、ODA2和ODA3。DPPH自由基清除率根据如下公式计算:
DPPH·清除率=(ODA2+ODA3-ODA1)/ODA2×100%。
⑥实验结果列于表4中。
表4革兰氏阳性杆菌发酵液上清对DPPH·自由基的清除率
如上表4所示,革兰氏阳性杆菌LMLC4560-1发酵得到的上清液对DPPH·自由基具有较高的清除率,达到90.35%。
实施例6菌株鉴定
将获得的革兰氏阳性菌LMLC4560-1菌株接种到BL固体培养基上划线培养,观察菌落形态。在BL培养基上,菌落呈淡黄色,扁平圆形,边缘整齐,不透明,1.5mm±0.3mm。菌落形态如图2。
挑取单菌落溶于1ml无菌水,在显微镜下观察其细胞形态。鉴定结果显示:得到的革兰氏阳性菌LMLC4560-1菌株不运动,无芽孢,无鞭毛,无荚膜,菌体呈一端钝圆、一端较小的短杆状,成对存在。菌株形态特征如图3。
对LMLC4560-1菌株进行16SrDNA测序及菌种鉴定。采用的引物为:正向引物27F:5'-AGAGTTTGATCATGGCTCAG-3';反向引物1492R:5'-ACGGCTACCTTGTTACGACTT-3'。PCR反应体系是:25μlMix;1μl样品;20μl ddH2O;2μl 27F;2μl 1492R;PCR反应温度:98℃,2min;98℃,10s,58℃,10s,72℃,7s,35x;72℃,2min;4℃保存。对PCR产物剂型测序,测得的LMLC4560-1菌株16SrDNA基因序列SEQ ID NO.2如下所示:
将LMLC4560-1菌株命名为食窦魏斯氏菌LMLC4560-1菌株(Weissella cibariaLMLC4560-1),并对其进行保藏,该食窦魏斯氏菌LMLC4560-1菌株(Weissella cibariaLMLC4560-1)于2021年7月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCCNO:M 2021811。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
序列表
<110> 湖北安琪生物集团有限公司
<120> 一株具有降胆固醇性能的乳酸菌及其应用
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1386
<212> DNA
<213> 植物乳杆菌(Lactobacillus plantarum )
<400> 1
ctaatacatg caagtcgaac gaactctggt attgattggt gcttgcatca tgatttacat 60
ttgagtgagt ggcgaactgg tgagtaacac gtgggaaacc tgcccagaag cgggggataa 120
cacctggaaa cagatgctaa taccgcataa caacttggac cgcatggtcc gagcttgaaa 180
gatggcttcg gctatcactt ttggatggtc ccgcggcgta ttagctagat ggtggggtaa 240
cggctcacca tggcaatgat acgtagccga cctgagaggg taatcggcca cattgggact 300
gagacacggc ccaaactcct acgggaggca gcagtaggga atcttccaca atggacgaaa 360
gtctgatgga gcaacgccgc gtgagtgaag aagggtttcg gctcgtaaaa ctctgttgtt 420
aaagaagaac atatctgaga gtaactgttc aggtattgac ggtatttaac cagaaagcca 480
cggctaacta cgtgccagca gccgcggtaa tacgtaggtg gcaagcgttg tccggattta 540
ttgggcgtaa agcgagcgca ggcggttttt taagtctgat gtgaaagcct tcggctcaac 600
cgaagaagtg catcggaaac tgggaaactt gagtgcagaa gaggacagtg gaactccatg 660
tgtagcggtg aaatgcgtag atatatggaa gaacaccagt ggcgaaggcg gctgtctggt 720
ctgtaactga cgctgaggct cgaaagtatg ggtagcaaac aggattagat accctggtag 780
tccataccgt aaacgatgaa tgctaagtgt tggagggttt ccgcccttca gtgctgcagc 840
taacgcatta agcattccgc ctggggagta cggccgcaag gctgaaactc aaaggaattg 900
acgggggccc gcacaagcgg tggagcatgt ggtttaattc gaagctacgc gaagaacctt 960
accaggtctt gacatactat gcaaatctaa gagattagac gttcccttcg gggacatgga 1020
tacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa 1080
cgagcgcaac ccttattatc agttgccagc attaagttgg gcactctggt gagactgccg 1140
gtgacaaacc ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgacctgggc 1200
tacacacgtg ctacaatgga tggtacaacg agttgcgaac tcgcgagagt aagctaatct 1260
cttaaagcca ttctcagttc ggattgtagg ctgcaactcg cctacatgaa gtcggaatcg 1320
ctagtaatcg cggatcagca tgccgcggtg aatacgttcc cgggccttgt acacaccgcc 1380
cgtcac 1386
<210> 2
<211> 1426
<212> DNA
<213> 食窦魏斯氏菌(Weissella cibaria)
<400> 2
aggttacccc accggctttg ggtgttacaa actctcatgg tgtgacgggc ggtgtgtaca 60
agacccggga acgtattcac cgcggcgtgc tgatccgcga ttactagcga ttccgacttc 120
atgtaggcga gttgcagcct acaatccgaa ctgagacgta ctttaagaga ttagctcacc 180
ctcgcgggtt ggcaactcgt tgtatacgcc attgtagcac gtgtgtagcc caggtcataa 240
ggggcatgat gatttgacgt catccccacc ttcctccggt ttgtcaccgg cagtctcact 300
agagtgccca actaaatgct ggcaactagt aataagggtt gcgctcgttg cgggacttaa 360
cccaacatct cacgacacga gctgacgaca accatgcacc acctgtcacc ttgtccccga 420
agggaacgct ccatctctgg agttgtcaag ggatgtcaag acctggtaag gttcttcgcg 480
ttgcttcgaa ttaaaccaca tgctccaccg cttgtgcggg tccccgtcaa ttcctttgag 540
tttcaacctt gcggtcgtac tccccaggcg gagtgcttaa tgcgttagct gcggcactta 600
agggcggaaa ccctcaaaca cctagcactc atcgtttacg gcgtggacta ccagggtatc 660
taatcctgtt tgctacccac actttcgagc ctcaacgtca gttacagwcc agaragccgc 720
cttcgccact ggtgttcytc catatatcta cgcatttcac cgctacacat ggaattccac 780
tctcctcttc tgcactcaag tctcccagtt tccaatgacc ctccccggtt gagccggggg 840
ctttcacatc agacttaaga aaccgcctgc gctcgcttta cgcccaataa atccggacaa 900
cgcttgccac ctacgtatta ccgcggctgc tggcacgtag ttagccgtgg ctttctggtt 960
agataccgtc aagggatgaa cagttactct catccttgtt cttctctaac aacagagttt 1020
tacgatccga aaaccttctt cactcacgcg gcgttgctcg gtcagacttt cgtccattgc 1080
cgaagattcc ctactgctgc ctcccgtagg agtttgggcc gtgtctcagt cccaatgtgg 1140
ccgatcaccc tctcaggtcg gctatgcatc gtagccttgg tgagccgtta cctcaccaac 1200
tagctaatgc accgcgggtc catccatcag cgacacccga aagcgccttt caaatcaaaa 1260
ccatgcggtt tcgattgtta tacggtatta gcacctgttt ccaagtgtta tccccttctg 1320
atgggcaggt tacccacgtg ttactcaccc gttcgccact cttctttttc cggtggagca 1380
agctccggtg gaaaaagaag cgtacgactg catgtattag cacgcc 1426
Claims (12)
1.一株食窦魏斯氏菌菌株,所述食窦魏斯氏菌菌株为食窦魏斯氏菌LMLC4560-1菌株(Weissella cibaria LMLC4560-1),其于2021年7月2日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为CCTCC NO:M 2021811。
2.根据权利要求1所述的食窦魏斯氏菌菌株,其特征在于,所述食窦魏斯氏菌LMLC4560-1菌株的16S rDNA基因序列如SEQ ID NO.2所示。
3.一种食窦魏斯氏菌菌剂的发酵制备方法,其特征在于,所述方法包含如下步骤:培养权利要求1或2所述的食窦魏斯氏菌菌株。
4.根据权利要求3所述的制备方法,其特征在于,所述制备方法包括如下步骤:
(1)将权利要求1或2所述的食窦魏斯氏菌菌株放大培养;
(2)将步骤(1)得到的产物加入到液体培养基中,在26-32℃条件下发酵培养。
5.一种菌剂,其特征在于,所述菌剂通过权利要求3或4所述的发酵制备方法得到。
6.权利要求1或2所述的食窦魏斯氏菌菌株或权利要求5所述的菌剂发酵获得的发酵产物。
7.根据权利要求6所述的发酵产物,其特征在于,所述发酵产物具有降胆固醇性能;
优选地,所述发酵产物具有自由基清除能力。
8.根据权利要求6所述的发酵产物,其特征在于,所述发酵产物作用于胆固醇,胆固醇的降低率大于87%;
或者,优选地,所述发酵产物对DPPH·自由基清除率大于90%。
9.权利要求6-8任意一项所述的发酵产物的制备方法,其特征在于,所述制备方法通过包括培养权利要求1或2所述的食窦魏斯氏菌菌株或权利要求5所述的菌剂制备得到。
10.根据权利要求9所述的制备方法,其特征在于,所述制备方法包括如下步骤:
(1)将权利要求1或2所述的食窦魏斯氏菌菌株或权利要求5所述的菌剂放大培养;
(2)将步骤(1)得到的产物加入到液体培养基中,在26-32℃条件下发酵培养得到所述发酵产物。
11.权利要求1或2所述的食窦魏斯氏菌菌株或权利要求5所述的菌剂在降低胆固醇、降脂、减肥、抗氧化或抗衰老产品中的应用。
12.根据权利要求11所述的应用,其特征在于,所述产品为食品、化妆品、保健品或药品。
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