CN111690571B - 一株可清除丙烯酰胺的植物乳杆菌及其应用 - Google Patents
一株可清除丙烯酰胺的植物乳杆菌及其应用 Download PDFInfo
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- CN111690571B CN111690571B CN202010671250.3A CN202010671250A CN111690571B CN 111690571 B CN111690571 B CN 111690571B CN 202010671250 A CN202010671250 A CN 202010671250A CN 111690571 B CN111690571 B CN 111690571B
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Abstract
本发明公开了一株可清除丙烯酰胺的植物乳杆菌及其应用,属于微生物技术领域以及医药技术领域。本发明提供了一株植物乳杆菌(Lactobacillus plantarum)CCFM1124,此植物乳杆菌CCFM1124能够清除丙烯酰胺,具体体现在:将此植物乳杆菌CCFM1124添加至丙烯酰胺溶液中培养2h,即可使丙烯酰胺溶液中丙烯酰胺的清除率达28.8%,可见,植物乳杆菌(Lactobacillus plantarum)CCFM1124在制备丙烯酰胺清除剂中具有巨大的应用前景。
Description
技术领域
本发明涉及一株可清除丙烯酰胺的植物乳杆菌及其应用,属于微生物技术领域以及医药技术领域。
背景技术
丙烯酰胺(CAS号79-06-1)为无色透明片状晶体,无臭,有毒。其相对密度1.122,熔点为84~85℃。溶于水、乙醇,微溶于苯、甲苯。极易升华,易聚合。固体在室温下稳定,在熔融时可猛烈聚合。
由于丙烯酰胺是一个具有亲电基团的有机小分子,水溶性极强,因此,其可通过皮肤、黏膜、呼吸道、胃肠道等进入人体。例如,食物中的丙烯酰胺可通过肠道被人体完整的吸收,空气中的丙烯酰胺约25%可通过皮肤被人体吸收。由于丙烯酰胺具有神经毒性、免疫毒性、生殖毒性和致癌性,这些被吸收的丙烯酰胺通过血液循环系统广泛分布于人体各个组织时,会对人体造成严重的损害,因此,为保持健康,人群需避免暴露于存在丙烯酰胺的环境中。
但是,丙烯酰胺在环境中是广泛存在的,其来源主要为烹饪不当的食品,尤其是油炸膨化食品。可见,人们在日常生活中几乎无法避免的会暴露于存在丙烯酰胺的环境中。2011年,FAO/WHO食品添加剂联合专家委员会(Joint FAO/WHO Expert Committee on FoodAdditives,JECFA)对除非洲以外世界范围内8个代表国家中丙烯酰胺膳食摄入量进行评估,结果表明普通人群的日摄入量平均约为1μg/(kgbw·d),最高摄入量约为4μg/(kgbw·d)。英国最新公布的日摄入量为0.61μg/(kgbw·d),法国为0.43μg/(kgbw·d),而中国在最新膳食研究中得出的摄入量为0.319μg/kgbw·d)。
因此,急需找到一种可有效清除环境中丙烯酰胺的产品来避免人群遭到丙烯酰胺暴露的伤害。
发明内容
[技术问题]
本发明要解决的技术问题是提供一株可清除丙烯酰胺的植物乳杆菌(Lactobacillus plantarum)。
[技术方案]
为解决上述问题,本发明提供了一株植物乳杆菌(Lactobacillus plantarum)CCFM1124,所述植物乳杆菌CCFM1124保藏于广东省微生物菌种保藏中心,保藏编号为GDMCCNo:61018,保藏日期为2020年05月06日。
所述植物乳杆菌CCFM1124来源于新疆省昌吉军户农场的人群肠道内容物样本,该菌株经测序分析,其16SrDNA序列如SEQ ID NO.1所示,将测序得到的序列在GeneBank中进行核酸序列比对,结果显示菌株为植物乳杆菌,命名为植物乳杆菌CCFM1124。
所述植物乳杆菌CCFM1124的形态特征为:圆端直杆菌。
所述植物乳杆菌CCFM1124的菌落特征为:圆形白色凸起,表面光滑。
本发明还提供了上述植物乳杆菌在制备预防和/或治疗丙烯酰胺暴露的药品中的应用。
本发明的一种实施方式中,所述药品中,上述植物乳杆菌CCFM1124的活菌数为不低于1×106CFU/mL或1×106CFU/g。
本发明的一种实施方式中,所述药品含有上述植物乳杆菌CCFM1124、药物载体和/或药用辅料。
本发明的一种实施方式中,所述药物载体包含微囊、微球、纳米粒和/或脂质体。
本发明的一种实施方式中,所述药用辅料包含赋形剂和/或附加剂。
本发明的一种实施方式中,所述赋形剂包含溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、吸收剂、稀释剂、絮凝剂、反絮凝剂、助滤剂和/或释放阻滞剂。
本发明的一种实施方式中,所述附加剂包含微晶纤维素、羟丙基甲基纤维素和/或精制卵磷脂。
本发明的一种实施方式中,所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
本发明还提供了一种的产品,所述产品含有上述植物乳杆菌CCFM1124。
本发明的一种实施方式中,所述产品中,上述植物乳杆菌CCFM1124的活菌数为不低于1×106CFU/mL或1×106CFU/g。
本发明的一种实施方式中,所述产品为食品、药品或丙烯酰胺清除剂。
本发明的一种实施方式中,所述药品含有上述植物乳杆菌CCFM1124、药物载体和/或药用辅料。
本发明的一种实施方式中,所述药物载体包含微囊、微球、纳米粒和/或脂质体。
本发明的一种实施方式中,所述药用辅料包含赋形剂和/或附加剂。
本发明的一种实施方式中,所述赋形剂包含溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、吸收剂、稀释剂、絮凝剂、反絮凝剂、助滤剂和/或释放阻滞剂。
本发明的一种实施方式中,所述附加剂包含微晶纤维素、羟丙基甲基纤维素和/或精制卵磷脂。
本发明的一种实施方式中,所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
本发明的一种实施方式中,所述食品为保健食品;或所述食品为使用含有上述植物乳杆菌CCFM1124的发酵剂生产得到的乳制品、豆制品或果蔬制品;或所述食品为含有上述植物乳杆菌CCFM1124的饮料或零食。
本发明的一种实施方式中,所述发酵剂的制备方法为将上述植物乳杆菌CCFM1124接种到培养基中进行培养,得到培养液;将培养液离心,得到菌体;将菌体用生理盐水或缓冲液清洗后用冻干保护剂重悬,得到重悬液;将重悬液采用真空冷冻法进行冻干,得到发酵剂。
在本发明的一种实施方式中,所述冻干保护剂包含100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
本发明还提供了上述植物乳杆菌或上述产品在清除丙烯酰胺中的应用。
有益效果:
(1)本发明提供了一株植物乳杆菌(Lactobacillus plantarum)CCFM1124,此植物乳杆菌CCFM1124能够清除丙烯酰胺,具体体现在:将此植物乳杆菌CCFM1124添加至丙烯酰胺溶液中培养2h,即可使丙烯酰胺溶液中丙烯酰胺的清除率达28.8%,可见,植物乳杆菌(Lactobacillus plantarum)CCFM1124在制备丙烯酰胺清除剂中具有巨大的应用前景。
(2)本发明提供了一株植物乳杆菌(Lactobacillus plantarum)CCFM1124,此植物乳杆菌CCFM1124能够缓解丙烯酰胺暴露,具体体现在:将此植物乳杆菌CCFM1124添加至丙烯酰胺溶液中培养2h,即可使丙烯酰胺溶液中丙烯酰胺的清除率达28.8%,可见,植物乳杆菌(Lactobacillus plantarum)CCFM1124在制备预防和/或治疗丙烯酰胺暴露的产品(如食品或药品等)中具有巨大的应用前景。
生物材料保藏
一株植物乳杆菌(Lactobacillus plantarum)CCFM1124,分类学命名为Lactobacillus plantarum,已于2020年05月06日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCCNo:61018,保藏地址为广州市先烈中路100号大院59号楼5楼。
附图说明
图1:不同植物乳杆菌清除水中丙烯酰胺的清除率。
具体实施方式
下述实施例中涉及的脱脂奶粉购自纽瑞兹食品有限公司;下述实施例中涉及的丙烯酰胺标准品购自美国sigma公司;下述实施例中涉及的植物乳杆菌(Lactobacillusplantarum)CCFM8661的保藏编号为CGMCCNo.5494,记载于公开号为CN102586148A的专利申请文本中;下述实施例中涉及的植物乳杆菌(Lactobacillus plantarum)CCFM8610的保藏编号为CGMCCNo.6077,记载于公开号为CN102827796A的专利申请文本中;下述实施例中涉及的植物乳杆菌(Lactobacillus plantarum)CCFM382的保藏编号为CGMCCNo.9734,记载于公开号为CN104357349A的专利申请文本中。
下述实施例中涉及的培养基如下:
LFMATA固体培养基(g/L):蛋白胨10g/L、牛肉膏10g/L、酵母粉5g/L、吐温1mL/L、磷酸氢二钾3g/L、乙酸钠2g/L、柠檬酸二铵2g/L、七水硫酸镁0.1g/L、一水硫酸锰0.05g/L、碳源20g/L、万古霉素20×10-3g/L、链霉素0.256g/L、庆大霉素6.4×10-2g/L、L-半胱氨酸0.5g/L、琼脂18g/L。
MRS固体培养基(g/L):蛋白胨10g/L、牛肉膏10g/L、葡萄糖20g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K2PO4·3H2O2.6g/L、MgSO4·7H2O0.1g/L、MnSO40.05g/L、吐温801mL/L、琼脂20g/L、半胱氨酸氨酸盐0.5g/L,pH为6.8。
MRS液体培养基(g/L):蛋白胨10g/L、牛肉膏10g/L、葡萄糖20g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K2PO4·3H2O2.6g/L、MgSO4·7H2O0.1g/L、MnSO40.05g/L、吐温801mL/L、半胱氨酸氨酸盐0.5g/L,pH为6.8。
下述实施例中涉及的检测方法如下:
活菌数的检测方法:采用国标《GB4789.35-2016食品安全国家标准食品微生物学检测乳酸菌检测》。
下述实施例中涉及的植物乳杆菌菌体的制备方法如下:
将植物乳杆菌菌液划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于MRS液体培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到植物乳杆菌菌体。
实施例1:植物乳杆菌CCFM1124的获取
具体步骤如下:
1、分离纯化
(1)稀释涂布:以来源于新疆省昌吉军户农场的人群肠道内容物为样本,吸取0.5g保存于30%(v/v)甘油中的样本在无菌环境下加入装有4.5mL生理盐水的10mL离心管中,得到10-1稀释液,重复上述稀释步骤,依次得到10-2、10-3、10-4、10-5、10-6稀释液;
(2)涂布培养:分别吸取100μL步骤(1)获得的10-4、10-5、10-6三个梯度稀释液涂布于LFMATA固体培养基上,37℃培养48h,得到稀释涂布平板;
(3)一级纯化培养:取步骤(2)获得的菌落数在30~300区间的稀释涂布平板,在每个稀释涂布平板上随机挑选10个乳白色或白色,表面光滑,边缘整齐,大小不一的单菌落在MRS固体培养基上划线,37℃培养48h,得到单菌落;
(4)二级纯化培养:取步骤(3)获得的单菌落分别接种至MRS液体培养基中,37℃培养20h,得到菌液。
2、菌种鉴定
将分离纯化的各菌液所对应的各菌株编号后,参照教科书《微生物学》(沈萍,陈向东主编)中所记载的步骤进行菌株鉴定、革兰氏染色、生理生化等实验,选取具有植物乳杆菌典型特征的菌株,经实验获得一株菌,将此菌株命名为CCFM1124;
其中,菌株鉴定过程如下:
提取CCFM1124的基因组,将CCFM1124的16SrDNA进行扩增和测序(由上海生工生物工程股份有限公司完成),将测序分析得到的CCFM1124的16SrDNA序列(CCFM1124的16SrDNA序列如SEQ ID NO.1所示)在GenBank中进行比对,结果显示此菌株确为植物乳杆菌,命名为植物乳杆菌(Lactobacillus plantarum)CCFM1124;
植物乳杆菌(Lactobacillus plantarum)CCFM1124的形态特征为:圆端直杆菌;
植物乳杆菌(Lactobacillus plantarum)CCFM1124的菌落特征为:圆形白色凸起,表面光滑;
植物乳杆菌(Lactobacillus plantarum)CCFM1124的生理生化特征为:革兰氏阳性兼性厌氧菌,对多种抗生素敏感,耐酸耐胆盐。
实施例2:植物乳杆菌CCFM1124对丙烯酰胺的清除作用
具体步骤如下:
准确称取10mg的丙烯酰胺标准品溶于1mL无菌水中,充分混匀溶解后使用0.22μm微孔滤膜进行过滤除菌,得到浓度为10mg/mL的丙烯酰胺标准品母液;准确称取10mg/mL的丙烯酰胺标准品母液100μL用无菌水定容至100mL,得到浓度为10μg/mL的丙烯酰胺标准品母液;以不添加植物乳杆菌菌体的浓度为10μg/mL的丙烯酰胺标准品母液作为空白对照,将3×109CFU的植物乳杆菌QS6-12菌体、植物乳杆菌RS15-3菌体、植物乳杆菌CCFM1124菌体、植物乳杆菌VJLHD7-L1菌体、植物乳杆菌VJLHD7-L1菌体、植物乳杆菌VJLHD11-L1菌体、植物乳杆菌FJHLD57M1菌体、植物乳杆菌QHLJZD24-L1菌体、植物乳杆菌PCQYD1M3菌体、植物乳杆菌CCFM8661菌体、植物乳杆菌CCFM8610菌体、植物乳杆菌PCQDDK5M2菌体、植物乳杆菌FZJTZ16M7菌体、植物乳杆菌FHNMY13M5菌体、植物乳杆菌VJLHD14-L1菌体、植物乳杆菌FCQNA34M6菌体、植物乳杆菌PCQWS1M2菌体、植物乳杆菌FSCPS8-3菌体、植物乳杆菌QHLJZD16L2菌体、植物乳杆菌FGDLZ5M7菌体、植物乳杆菌DHuNHHMY9-L1菌体、植物乳杆菌DHuNHHMY10-L1菌体、植物乳杆菌DHuNHHMY13-L2菌体、植物乳杆菌CCFM382菌体、植物乳杆菌FSCPS8-3菌体、植物乳杆菌QHLJZD16L2菌体、植物乳杆菌FGDLZ5M7菌体、植物乳杆菌DHuNHHMY9-L1菌体、植物乳杆菌DHuNHHMY13-L2菌体依次分别与1mL浓度为10μg/mL的丙烯酰胺标准品母液混合后于37℃、150r/min的条件下共培养2h,得到培养液1~29;将培养液1~2912000rpm离心10min,取上清用0.22μm微孔滤膜进行过滤除菌,得到滤液1~29;其中,植物乳杆菌QS6-12、植物乳杆菌RS15-3、植物乳杆菌VJLHD7-L1、植物乳杆菌VJLHD7-L1、植物乳杆菌VJLHD11-L1、植物乳杆菌FJHLD57M1、植物乳杆菌QHLJZD24-L1、植物乳杆菌PCQYD1M3、植物乳杆菌PCQDDK5M2、植物乳杆菌FZJTZ16M7、植物乳杆菌FHNMY13M5、植物乳杆菌VJLHD14-L1、植物乳杆菌FCQNA34M6、植物乳杆菌PCQWS1M2、植物乳杆菌FSCPS8-3、植物乳杆菌QHLJZD16L2、植物乳杆菌FGDLZ5M7、植物乳杆菌DHuNHHMY9-L1、植物乳杆菌DHuNHHMY10-L1、植物乳杆菌DHuNHHMY13-L2、植物乳杆菌FSCPS8-3、植物乳杆菌QHLJZD16L2、植物乳杆菌FGDLZ5M7、植物乳杆菌DHuNHHMY9-L1、植物乳杆菌DHuNHHMY13-L2均是从人或动物粪便以及发酵制品中筛选得到的。
检测滤液1~29中丙烯酰胺的含量,并根据公式:清除率(%)=[(空白对照中丙烯酰胺的含量-滤液中丙烯酰胺的含量)/空白对照中丙烯酰胺的含量]×100%,计算滤液1~29中丙烯酰胺的清除率,检测结果见图1。
由图1可知,滤液3中苯并芘的含量最低(低至7.1μg)、苯并芘的清除率最高(高达28.8%)。可见,植物乳杆菌(Lactobacillus plantarum)CCFM1124可有效清除二甲基亚砜中的丙烯酰胺。
实施例3:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备菌粉,菌粉的具体制备过程如下:
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;将菌悬液在温度37℃下预培养60min后冻干,得到植物乳杆菌CCFM1124菌粉;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基;
保护剂的成分包含:100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
实施例4:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备牛乳,发酵乳的具体制备过程如下:
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;将菌悬液在温度37℃下预培养60min后冻干,得到植物乳杆菌CCFM1124菌粉;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基;
保护剂的成分包含:100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
将脱脂奶在95℃热杀菌20min后冷却至4℃,得到原料;在原料中添加植物乳杆菌CCFM1124菌粉至浓度为不低于1×106CFU/mL,得到牛乳。
实施例5:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备豆奶,豆奶的具体制备过程如下:
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;将菌悬液在温度37℃下预培养60min后冻干,得到植物乳杆菌CCFM1124菌粉;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基;
保护剂的成分包含:100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
将大豆在温度80℃下浸泡2h后去除大豆皮,得到去皮大豆;将去皮大豆沥去浸泡水后加沸水磨浆,得到豆浆;将豆浆在高于80℃的温度条件下保温12min,得到熟豆浆;将熟豆浆用150目筛网过滤后离心分离,得到粗豆奶;将粗豆奶加热到温度140~150℃后迅速导入真空冷却室进行抽真空,使得粗豆奶中的异味物质随着水蒸汽迅速排出,得到熟豆奶;将熟豆奶降温至约37℃后在熟豆奶中添加植物乳杆菌CCFM1124菌粉至浓度为不低于1×106CFU/mL,得到豆奶。
实施例6:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备果蔬饮料,果蔬饮料的具体制备过程如下:
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;将菌悬液在温度37℃下预培养60min后冻干,得到植物乳杆菌CCFM1124菌粉;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基;
保护剂的成分包含:100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
将新鲜水果和蔬菜洗净后榨汁,得到果蔬汁;将果蔬汁在温度140℃下高温热杀菌2秒,得到杀菌后的果蔬汁;将杀菌后的果蔬汁降温至约37℃后在杀菌后的果蔬汁中添加植物乳杆菌CCFM1124菌粉至浓度为不低于1×106CFU/mL,得到果蔬饮料。
实施例7:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备胶囊制品,胶囊制品的具体制备过程如下
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;菌悬液添加至浓度为30g/L的海藻酸钠溶液中至浓度为2×109CFU/mL后,充分搅拌,使得植物乳杆菌CCFM1124的细胞均匀地分散于海藻酸钠溶液中,得到混合液;将混合液挤压到浓度为20g/L的氯化钙溶液中形成胶粒;待形成的胶粒静止固化30min后,过滤收集胶粒;将收集得到的胶粒进行冷冻干燥48h,得到粉剂;将粉剂装入到药用胶囊中,得到胶囊制品;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基。
实施例8:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备发酵乳,发酵乳的具体制备过程如下:
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;将菌悬液在温度37℃下预培养60min后冻干,得到植物乳杆菌CCFM1124菌粉;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基;
保护剂的成分包含:100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
将植物乳杆菌CCFM1124菌粉与商业干粉发酵剂保加利亚乳杆菌和商业干粉发酵剂嗜热链球菌按照质量比1:1:1的比例混合,得到发酵剂;将糖添加至鲜奶中至浓度为50g/L,得到混合液;将混合液在65℃、20MPa的条件下进行均质后在95℃下保温杀菌5min,得到发酵原料;将发酵原料降温至35℃后以0.03%(v/v)的接种量将发酵剂接种至发酵原料中,于35℃下保温发酵16h,得到发酵乳;将发酵乳于42℃下放置4h进行凝乳后,在4℃下冷藏24h进行后熟,得到发酵乳成品。
实施例9:植物乳杆菌CCFM1124的应用
具体步骤如下:
植物乳杆菌CCFM1124可用于制备片剂,片剂的具体制备过程如下:
植物乳杆菌CCFM1124划线于MRS固体培养基上,37℃条件下培养48h,得到单菌落;挑取单菌落接种于MRS液体培养基中,37℃条件下培养18h进行活化,连续活化两代,得到活化液;将活化液按2%(v/v)的接种量接种于培养基中,37℃条件下培养18h,得到菌液;将菌液经5000g离心15min,得到菌泥;将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×1010CFU/mL,得到菌悬液;将菌悬液在温度37℃下预培养60min后冻干,得到植物乳杆菌CCFM1124菌粉;
其中,培养基的制备方法为:使用以培养基总重量计87.7%的水将10%酶水解脱脂乳、0.5%葡萄糖、1.5%胰蛋白胨与0.3%酵母浸膏溶解,然后调整其pH为6.8,得到培养基;
保护剂的成分包含:100g/L脱脂奶粉、30mL/L甘油、100g/L麦芽糊精、150g/L海藻糖和10g/LL-谷氨酸钠。
称取植物乳杆菌CCFM1124菌粉25.7重量份、淀粉55.0重量份、纤维素衍生物4.5重量份、羧甲基淀粉钠12.0重量份、滑石粉0.8重量份、蔗糖1.0重量份与水1.0重量份,得到原材料;将原材料混合,得到湿颗粒;将湿颗粒用中南制药机械厂的压片机进行压片后使用青州市益康中药机械有限公司的小型药物干燥机进行干燥,得到片剂。
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
序列表
<110> 江南大学
<120> 一株可清除丙烯酰胺的植物乳杆菌及其应用
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 467
<212> DNA
<213> 植物乳杆菌
<400> 1
agcatgctga caacccgtta tttagatatg cgatatgctt attctagcaa ccgctttaag 60
acggacacga tccagacctg ctagttgtct cccccaatcg acgacccact ctcccaacca 120
aaataccgca tgtttttatt taacaattca ccaaccttaa cggagaaaat ccaactaatt 180
tcctgactaa gtacagtccc gaacccccga cagtttgacg actagtccac aacgacaacc 240
acataccatt cagctatggc gtaatactaa cccagtaacg ttttagtagc gatcgtcatg 300
ggctactttt ggcataaacc tactatagta atacttaatt aatcgacgcc ttcgttaacc 360
gaccacaatt gactacacga tgcgcaactt ctatcaatta tctgctttac cgaaaaataa 420
ttagtcagtg ccacgtctcc aagcaataaa atgactagtt aagtgtc 467
Claims (8)
1.一株植物乳杆菌(Lactobacillus plantarum),其特征在于,所述植物乳杆菌保藏于广东省微生物菌种保藏中心,保藏编号为GDMCC No:61018,保藏日期为2020年05月06日。
2.权利要求1所述植物乳杆菌在制备预防和/或治疗丙烯酰胺暴露诱发的疾病的药品中的应用。
3.一种产品,其特征在于,所述产品含有权利要求1所述植物乳杆菌;所述产品为食品、药品或丙烯酰胺清除剂。
4.如权利要求3所述的产品,其特征在于,所述产品中,权利要求1所述植物乳杆菌的活菌数为不低于1×106CFU/mL或1×106CFU/g。
5.如权利要求3所述的产品,其特征在于,所述药品含有权利要求1所述植物乳杆菌、以及药物载体和/或药用辅料。
6.如权利要求5所述的产品,其特征在于,所述药物载体为微囊、微球、纳米粒和/或脂质体。
7.如权利要求6所述的产品,其特征在于,所述药用辅料为赋形剂和/或附加剂。
8.如权利要求7所述的产品,其特征在于,所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
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