CN110878270B - 一株副干酪乳杆菌副干酪亚种及其应用 - Google Patents
一株副干酪乳杆菌副干酪亚种及其应用 Download PDFInfo
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- CN110878270B CN110878270B CN201911331794.9A CN201911331794A CN110878270B CN 110878270 B CN110878270 B CN 110878270B CN 201911331794 A CN201911331794 A CN 201911331794A CN 110878270 B CN110878270 B CN 110878270B
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Abstract
本发明公开了一株副干酪乳杆菌副干酪亚种,所述菌株保藏名称为副干酪乳杆菌副干酪亚种BB4,于2018年9月10日保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为:CGMCC No.16455。本发明还公开了该副干酪乳杆菌副干酪亚种在制备抑制创伤弧菌药物、保健食品或普通食品中的应用,以及在降解偶氮类人工合成色素中的应用。本发明提供的副干酪乳杆菌副干酪亚种BB4以及利用该菌株制备的发酵酸奶能够显著抑制致病菌创伤弧菌的生长;该菌株能够显著显著降解偶氮类人工合成食用色素。因此,本发明提供的副干酪乳杆菌副干酪亚种BB4具有较高的社会应用价值。
Description
技术领域
本发明涉及微生物菌株及其应用技术领域,具体涉及一株副干酪乳杆菌副干酪亚种及其应用。
背景技术
偶氮类合成色素因合成工艺简单、成本低、染色性能突出等优点,被广泛的应用在食品、药品和化妆品等领域。其种类有很多,熟知的有柠檬黄、日落黄、柠檬黄、苋菜红、胭脂红和诱惑红等。目前已有不少研究发现了偶氮类色素的毒性作用,如使肝细胞生长异常,影响酯酶同工酶的表达,以及致突变作用等,因此多数偶氮色素已被禁止添加到食物中。我国2014年最新颁布的《食品添加剂使用标准(GB276 0-2014)》中对允许使用的柠檬黄、日落黄、柠檬黄、苋菜红、胭脂红和诱惑红等11种偶氮类色素在食品中的限量及使用范围做了明确的规定,严禁超量或超范围添加。
副干酪乳杆菌(Lactobacillus paracasei)是我国批准的可用于普通食品的菌种之一。副干酪乳杆菌广泛存在于人体口腔及肠道,发酵乳制品和植物原料发酵物等诸多环境中,它具有干酪乳杆菌属调节肠道和增强免疫力等益生功能,其代谢产生的细菌素具有优良的抑菌性能。目前,副干酪乳杆菌的益生功能及其在食品、医疗保健等领域发挥的作用已受到食品届内学者的高度重视。副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)属于副干酪乳杆菌属中的一个亚种,目前为止,行业内未见将副干酪乳杆菌副干酪亚种开发用于抑制特定致病菌创伤弧菌的药物、保健食品或普通食品的相关报道,也未见将副干酪乳杆菌副干酪亚种用于降解偶氮类人工合成食用色素的相关报道。
发明内容
本发明的目的是提供一株副干酪乳杆菌副干酪亚种及其应用,开发了一株能够显著降解偶氮类人工合成食用色素,并对创伤弧菌抑制能力较强的菌株。
本发明的目的是通过以下技术方案实现的:一株副干酪乳杆菌副干酪亚种,所述菌株保藏名称为副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4,于2018年9月10日保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为:CGMCC No.16455。保藏单位地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101。
本发明提供的副干酪乳杆菌副干酪亚种BB4分离自人类母乳,菌落特征为:在MRS平板培养基上培养24~48 h后,菌落呈圆形,乳白色,表面光滑、***,边缘整齐,不透明,菌体呈杆状,大小为0.3~0.6 × 1.0~3.0 μm;能够生长的pH值为4.0~9.0,最适生长pH为5.5,能够生长的温度为20~40℃,最适生长温度37.5℃;能够利用的糖类包括:核糖、D-半乳糖、D-葡萄糖、D-果糖、D-甘露糖、甘露醇、山梨醇、N-乙酰葡萄糖胺、苦杏仁苷、熊果苷、七叶苷柠檬酸铁、水杨苷(柳醇)、D-纤维二糖、D-麦芽糖、D-乳糖、D-密二糖、D-蔗糖、海藻糖、菊粉、D-松三糖、D-棉子糖、D-龙胆二糖、D-塔格糖和葡萄糖酸盐;能够分泌白氨酸芳胺酶、缬氨酸芳胺酶、胱氨酸芳胺酶、β-葡萄糖苷酶、萘酚-AS-BI-磷酸水解酶、β-半乳糖苷酶、α-葡萄糖苷酶、酸性磷酸酶、酯酶和类脂肪酶。该菌株及其发酵酸奶对致病菌创伤弧菌的生长具有显著的抑制作用;该菌株能够明显降解偶氮类人工合成食用色素。
本发明所述的副干酪乳杆菌副干酪亚种在制备抑制创伤弧菌药物、保健食品或普通食品中的应用;所述药物、保健食品或普通食品中所含副干酪乳杆菌副干酪亚种BB4的活菌数为1.0×106-5.0×1011cfu/mL或1.0×106-5.0×1011cfu/g,优选活菌数为2.0×106-2.0×108cfu/mL或2.0×106-2.0×108cfu/g,最优选活菌数为2.0×107 cfu/mL或2.0×107cfu/g。
本发明所述的副干酪乳杆菌副干酪亚种在降解偶氮类人工合成食用色素的应用,具体是将所述副干酪乳杆菌副干酪亚种BB4添加到含有偶氮类人工合成食用色素的培养基中,所述基质中所含副干酪乳杆菌副干酪亚种BB4的活菌数为1.0×106 ~5.0×1010 cfu/mL或1.0×106 ~5.0×1010 cfu/g,优选活菌数为活菌数为1.0 ×10 6 ~ 1.0×109 cfu/mL或1.0 ×10 6 ~ 1.0×109 cfu/g,最优选活菌数1.0×109 cfu/mL或1.0×109 cfu/g。
本发明的有益效果在于:本发明提供的副干酪乳杆菌副干酪亚种BB4分离自人类母乳,安全性高,该菌株能够显著抑制致病菌创伤弧菌的生长;该菌株还能够显著降解偶氮类人工合成食用色素,为偶氮类人工合成食用色素的降解制剂提供了新的选择。因此,本发明提供的副干酪乳杆菌副干酪亚种BB4具有较高的社会应用价值。
附图说明
图1为副干酪乳杆菌副干酪亚种BB4的菌落形态。
图2为副干酪乳杆菌副干酪亚种BB4的扫描电镜图。
图3为副干酪乳杆菌副干酪亚种BB4在不同温度下的生长曲线。
图4为副干酪乳杆菌副干酪亚种BB4在不同pH值下的生长曲线。
具体实施方式
以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,若未特别指明,实施例中的培养基及实验条件均为常规培养基和实验条件。实施例中所用的试验材料、试剂等,如无特殊说明,均可从商业途径得到。
实施例1 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4的分离与鉴定
1、菌株分离
从一名来自中国的健康妇女获取母乳,将获取的新鲜初乳置于超净工作台中用无菌的PBS溶液(浓度为10 mM)稀释,将未稀释的乳汁和稀释5倍的乳汁分别分配到BBL平板培养基中,在37℃下恒温厌氧培养,直至菌落出现,用接种针挑取边缘生长良好的细菌,接种于新的BBL平板培养基上,待新接种的菌长成菌落后,再挑取其边缘的内生菌划线培养,如此反复得到纯化,直到得到单菌落,命名其为菌株BB4。
其中BBL平板培养基配方为:蛋白胨15.0 g/L、酵母粉2.0 g/L、葡萄糖20.0 g/L、可溶性淀粉0.5 g/L、NaCl 5.0 g/L、半胱氨酸 10.0 mL/L、西红柿浸出液 400 mL/L、吐温80 1.0 mL/L、肝提取液 80.0 mL/L、琼脂20.0 g/L、蒸馏水520.0 mL/L,pH值为6.0,在121℃灭菌20 min。
2、菌株鉴定
(1)菌落及菌体的形态观察:将菌株BB4在MRS平板培养基上进行划线培养,在37℃恒温厌氧培养24~48 h后,菌落呈圆形,乳白色,表面光滑、***,边缘整齐,不透明,菌落形态见图1。在扫描电子显微镜下观察,菌体呈杆状,大小为0.3~0.6 × 1.0~3.0 μm其扫描电镜图为图2。
其中MRS平板培养基配方为:胰蛋白胨10.0 g/L、牛肉粉5.0 g/L、酵母粉4.0 g/L、葡萄糖20.0 g/L、吐温80 1.0 mL/L、K2HPO4•3H2O 1.5 g/L、无水乙酸钠3.0 g/L、柠檬酸铵2.0 g/L、MgSO4•7H2O 0.2 g/L、MnSO4•H2O 0.04 g/L,琼脂15.0 g/L。pH值为6.0,在121℃灭菌20 min。
(2)菌株BB4的生长特征观察:将菌株BB4(菌体浓度约为1.0×107)按体积比3.0%的接种量接种于MRS液体培养基中,置于不同温度(10℃-50℃)下厌氧培养24 h,发现菌株在10℃或50℃下基本不能生长,在30℃~40℃环境下生长良好,最适生长温度为37.5℃(见图3);置于不同初始pH值(2.0-12.0)环境下厌氧培养24 h,发现菌株在pH值范围为3.0~11.0的环境中菌株能够生长,最适生长pH值为5.5(见图4)。
(3)菌株BB4的分子生物学鉴定:采用细菌全基因组快速抽提试剂盒,提取菌株BB4的全基因组,通过选用细菌16S rDNA通用引物27F和1492R进行PCR,然后测序分析。测序结果经BLAST比对,鉴定该菌株为副干酪乳杆菌副干酪亚种,基因序列如SEQ ID NO : 1所示,命名为:副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4,于2018年9月10日保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为:CGMCC No.16455。保藏单位地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101。
实施例2 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4对糖类的利用性
应用API50CHL 试剂盒(Biomerieux,法国)检测实施例1中分离的菌株BB4对糖类的利用性,结果(表1)显示:本发明副干酪乳杆菌副干酪亚种BB4能够利用核糖、D-半乳糖、D-葡萄糖、D-果糖、D-甘露糖、卫矛醇、甘露醇、山梨醇、N-乙酰葡萄糖胺、苦杏仁苷、熊果苷、七叶苷柠檬酸铁、水杨苷(柳醇)、D-纤维二糖、D-麦芽糖、D-乳糖、D-密二糖、D-蔗糖、海藻糖、菊粉、D-松三糖、D-棉子糖、D-龙胆二糖、D-塔格糖、D-土伦糖和葡萄糖酸盐。
表1 应用API50CHL 试剂盒检测菌株BB4对糖类的利用性结果
备注:+表示阳性结果,说明菌株能够利用受试底物;-表示阴性结果,说明菌株不能利用受试底物。
实施例3 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4的酶活性
应用APIZYM 试剂盒(Biomerieux,法国)检测实施例1 中分离的菌株BB4的酶活性,结果(表2)显示:本发明副干酪乳杆菌副干酪亚种BB4能够分泌白氨酸芳胺酶、缬氨酸芳胺酶、胱氨酸芳胺酶、萘酚-AS-BI-磷酸水解酶、β-半乳糖苷酶、α-葡萄糖苷酶、酸性磷酸酶、酯酶和类脂肪酶。
表2 应用APIZYM 试剂盒检测BB4的酶活性结果
备注:分值0-1表示为阴性反应;分值2表示反应微弱;分值为3-5表示阳性反应;颜色越深,分值越高。
实施例4 副干酪乳杆菌副干酪亚种BB4发酵酸奶的制备
将实施例1中所述副干酪乳杆菌副干酪亚种BB4作为发酵菌株接种于液体乳培养基中。所述液体乳培养基配方为:每100 mL蒸馏水中添加奶粉12 g、白砂糖8 g。经100 ℃水浴加热30 min后,冷却至室温,置于超净工作台,吸取驯化后的副干酪乳杆菌副干酪亚种BB4种子液(浓度约为1.0×107cfu/mL)按接种体积比10.0%的接种量接种于上述液体乳培养基中,充分混匀,经37 ℃发酵12 h,得到副干酪乳杆菌副干酪亚种BB4发酵酸奶,测其活菌数约为3.0×107 cfu/mL。
实施例5 副干酪乳杆菌副干酪亚种BB4及其发酵酸奶对创伤弧菌生长的抑制试验
实验方法:以创伤弧菌为指示菌,将指示菌株(浓度约为:1.0×106 cfu/mL)分别涂布于LB琼脂平板上,用无菌打孔器在平板上打直径为6 mm的加样孔,每组设置3个重复;在孔中加入100 μL的BB4菌液(4.0×107 cfu/mL)、菌液的培养上清液以及实施例3中所述的发酵酸奶,于37℃生化培养箱中培养24 h。观察并记录抑菌圈大小。
实验结果如下:
表3 本发明副干酪乳杆菌副干酪亚种BB4及其发酵酸奶对创伤弧菌的抗菌活性
表3结果显示,BB4菌液、培养上清液及其发酵酸奶对创伤弧菌的显抑菌圈大小分别为1.3 cm、1.3 cm和1.0 cm,表明副干酪乳杆菌副干酪亚种BB4、培养上清液及其发酵酸奶对创伤弧菌的生长具有明显的抑制效果。
实施例6 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4在降解日落黄色素中的应用
实验方法:将菌株BB4(菌体浓度约为1.0×107 cfu/mL)按体积比5%的接种量分别接种于MRS液体培养基中,置于37℃下厌氧培养24 h后,离心收集菌体沉淀,经适当稀释调整,使菌体浓度分别为1.0×109 cfu/mL、1.0×108 cfu/mL、1.0×107 cfu/mL和1.0×106cfu/mL,同时根据《食品添加剂使用标准(GB276 0-2014)》规定,日落黄色素在食品中的最大允许使用量为0.05 g/kg,设置日落黄色素在含BB4菌体的MRS液体培养基中的浓度分别为0.05、0.025和0.0125 g/L,于不同培养时间(3 h ~ 48 h)下取培养液,与1.5 g/L乙酸铵溶液按1:1比例稀释,在480 nm下测吸光值,按照下述公式计算降解率。
降解率(%)=(OD 0 h-OD不同培养时间)/OD 0 h ×100。
实验结果如下:
表4 不同培养时间下副干酪乳杆菌副干酪亚种BB4菌株对日落黄色素的降解效果
表4结果显示,副干酪乳杆菌副干酪亚种BB4菌株对日落黄色素具有非常明显的降解效果,在培养时间为3 h,菌体浓度为1.0×109 cfu/mL,日落黄色素浓度为0.025 g/L时,BB4菌株对日落黄色素的降解率已经高达73.69%;在培养时间为24 h时,1.0×106 cfu/mL ~1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.05 g/L浓度范围内的日落黄色素的降解率均在80%以上,最高达94.38%;在培养时间为36 h时,1.0×106 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.05 g/L浓度范围内的日落黄色素的降解率均在85%以上,最高达100%;在培养时间为48 h时,1.0×106 cfu/mL ~ 1.0×109cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.05 g/L浓度范围内的日落黄色素的降解率均在95%以上,最高达100%。
实施例7 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4在降解柠檬黄色素中的应用
实验方法:将菌株BB4(菌体浓度约为1.0×107 cfu/mL)按体积比5%的接种量分别接种于MRS液体培养基中,置于37℃下厌氧培养24 h后,离心收集菌体沉淀,经适当稀释调整,使菌体浓度分别为1.0×109 cfu/mL、1.0×108 cfu/mL、1.0×107 cfu/mL和1.0×106cfu/mL,同时根据《食品添加剂使用标准(GB276 0-2014)》规定,柠檬黄色素在食品中的最大允许使用量为0.05 g/kg,设置柠檬黄色素在含BB4菌体的MRS液体培养基中的浓度分别为0.05、0.025和0.0125 g/L,于不同培养时间(3 h ~ 48 h)下取培养液,与1.5 g/L乙酸铵溶液按1:1比例稀释,在426 nm下测吸光值,按照下述公式计算降解率。
降解率(%)=(OD 0 h-OD不同培养时间)/OD 0 h ×100。
实验结果如下:
表5 不同培养时间下副干酪乳杆菌副干酪亚种BB4菌株对柠檬黄色素的降解效果
表5结果显示,副干酪乳杆菌副干酪亚种BB4菌株对柠檬黄色素具有一定的降解效果,在培养时间为6 h,菌体浓度为1.0×109 cfu/mL,柠檬黄色素浓度为0.0125 g/L时,BB4菌株对柠檬黄色素的降解率高于50%,达54.44%;在培养时间为24 h时,1.0×108 cfu/mL ~1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.025 g/L 浓度范围内的柠檬黄色素的降解率达70%以上,最高达84.11;在培养时间为36 h时,1.0×108 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.025 g/L 浓度范围内的柠檬黄色素的降解率为90%左右,最高达94.60%;在培养时间为48 h时,1.0×106 cfu/mL ~ 1.0×109cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.025 g/L浓度范围内的柠檬黄色素的降解率均在95%以上,最高达100%。
实施例8 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4在降解苋菜红色素中的应用
实验方法:将菌株BB4(菌体浓度约为1.0×107 cfu/mL)按体积比5%的接种量分别接种于MRS液体培养基中,置于37℃下厌氧培养24 h后,离心收集菌体沉淀,经适当稀释调整,使菌体浓度分别为1.0×109 cfu/mL、1.0×108 cfu/mL、1.0×107 cfu/mL和1.0×106cfu/mL,同时根据《食品添加剂使用标准(GB276 0-2014)》规定,苋菜红色素在食品中的最大允许使用量为0.025 g/kg,设置苋菜红色素在含BB4菌体的MRS液体培养基中的浓度分别为0.025、0.0125和0.00625 g/L,于不同培养时间(3 h ~ 48 h)下取培养液,与1.5 g/L乙酸铵溶液按1:1比例稀释,在518 nm下测吸光值,按照下述公式计算降解率。
降解率(%)=(OD 0 h-OD不同培养时间)/OD 0 h ×100。
实验结果如下:
表6 不同培养时间下副干酪乳杆菌副干酪亚种BB4菌株对苋菜红色素的降解效果
表6结果显示,副干酪乳杆菌副干酪亚种BB4菌株对苋菜红色素具有极其显著的降解效果,在培养时间为3 h,菌体浓度为1.0×109 cfu/mL,苋菜红色素浓度为0.025 g/L时,BB4菌株对苋菜红色素的降解率已经高达82.02%;在培养时间为6 h时,1.0×108 cfu/m ~1.0×109 cfu/mL浓度范围内的BB4菌体对0.00625 g/L ~ 0.025 g/L浓度范围内的苋菜红色素的降解率均在85%以上,最高达98.87%;在培养时间为12 h时,1.0×106 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.00625 g/L ~ 0.0125 g/L浓度范围内的苋菜红色素的降解率均在95%以上,最高达100%;在培养时间为24 h、36 h和48 h时,1.0×106 cfu/mL~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.00625 g/L ~ 0.025 g/L浓度范围内的苋菜红色素的降解均已彻底,降解率均达100%。
实施例9 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4在降解胭脂红色素中的应用
实验方法:将菌株BB4(菌体浓度约为1.0×107 cfu/mL)按体积比5%的接种量分别接种于MRS液体培养基中,置于37℃下厌氧培养24 h后,离心收集菌体沉淀,经适当稀释调整,使菌体浓度分别为1.0×109 cfu/mL、1.0×108 cfu/mL、1.0×107 cfu/mL和1.0×106cfu/mL,同时根据《食品添加剂使用标准(GB276 0-2014)》规定,胭脂红色素在食品中的最大允许使用量为0.05g /kg,设置胭脂红色素在含BB4菌体的MRS液体培养基中的浓度分别为0.05、0.025和0.0125 g/L,于不同培养时间(3 h ~ 48 h)下取培养液,与1.5 g/L乙酸铵溶液按1:1比例稀释,在505 nm下测吸光值,按照下述公式计算降解率。
降解率(%)=(OD 0 h-OD不同培养时间)/OD 0 h ×100。
实验结果如下:
表7 不同培养时间下副干酪乳杆菌副干酪亚种BB4菌株对胭脂红色素的降解效果
表7结果显示,副干酪乳杆菌副干酪亚种BB4菌株对胭脂红色素具有不错的降解效果,在培养时间为6 h,1.0×108 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.05 g/L浓度范围内的胭脂红色素的降解率均在70%以上,最高达88.35%;在培养时间为12 h时,1.0×108 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.05 g/L浓度范围内的胭脂红色素的降解率均在85%以上,最高达100%;在培养时间为24 h时,1.0×108 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125g/L ~ 0.05 g/L浓度范围内的胭脂红色素的降解率均在90%以上,最高达100%;在培养时间为36 h时,1.0×106 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~0.05 g/L浓度范围内的胭脂红色素的降解率均在90%以上,最高达100%;在培养时间为48 h时,1.0×106cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.0125 g/L ~ 0.05 g/L浓度范围内的胭脂红色素的降解已经完全,达100%。
实施例10 副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4在降解诱惑红色素中的应用
实验方法:将菌株BB4(菌体浓度约为1.0×107 cfu/mL)按体积比5%的接种量分别接种于MRS液体培养基中,置于37℃下厌氧培养24 h后,离心收集菌体沉淀,经适当稀释调整,使菌体浓度分别为1.0×109 cfu/mL、1.0×108 cfu/mL、1.0×107 cfu/mL和1.0×106cfu/mL,同时根据《食品添加剂使用标准(GB276 0-2014)》规定,诱惑红色素在食品中的最大允许使用量为0.025 g/kg,设置诱惑红色素在含BB4菌体的MRS液体培养基中的浓度分别为0.025、0.0125和0.00625 g/L,于不同培养时间(3 h ~ 48 h)下取培养液,与1.5 g/L乙酸铵溶液按1:1比例稀释,在500 nm下测吸光值,按照下述公式计算降解率。
降解率(%)=(OD 0 h-OD不同培养时间)/OD 0 h ×100。
实验结果如下:
表8 不同培养时间下副干酪乳杆菌副干酪亚种BB4菌株对诱惑红色素的降解效果
表8结果显示,副干酪乳杆菌副干酪亚种BB4菌株对诱惑红色素具有极其显著的降解效果,在培养时间为3 h,菌体浓度为1.0×109 cfu/mL,诱惑红色素浓度为0.025 g/L时,BB4菌株对诱惑红色素的降解率已经高于50%;在培养时间为12 h时,1.0×108 cfu/m ~1.0×109 cfu/mL浓度范围内的BB4菌体对0.00625 g/L ~ 0.025 g/L浓度范围内的诱惑红色素的降解率均在90%以上,最高达100%;在培养时间为24 h时,1.0×106 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.00625 g/L ~ 0.0125 g/L浓度范围内的诱惑红色素的降解率均在95%以上,最高达100%;在培养时间为36 h和48 h时,1.0×106 cfu/mL ~ 1.0×109 cfu/mL浓度范围内的BB4菌体对0.00625 g/L ~ 0.025 g/L浓度范围内的诱惑红色素的降解均已彻底,降解率均达100%。
综上所述,本发明提供的副干酪乳杆菌副干酪亚种BB4对致病菌创伤弧菌的生长具有明显的抑制作用,该菌株对偶氮类人工合成使用色素(柠檬黄、日落黄、柠檬黄、苋菜红、胭脂红和诱惑红)具有显著的降解效果。鉴于副干酪乳杆菌副干酪亚种是我国批准的可用于普通食品的菌种之一,且分离自人类母乳,安全性很高,所以本发明提供的副干酪乳杆菌副干酪亚种BB4具有很高的实际应用价值。以上仅是本发明的若干个具体实施例。
本发明不限于以上实施例,还可以有许多变形。本领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形,均应认为是本发明的保护范围。
SEQUENCE LISTING
<110> 福建省农业科学院农业工程技术研究所
<120> 一株副干酪乳杆菌副干酪亚种及其应用
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1433
<212> DNA
<213> 人工序列
<400> 1
tgcagtcgaa cgagttctcg ttgatgatcg gtgcttgcac cgagattcaa catggaacga 60
gtggcggacg ggtgagtaac acgtgggtaa cctgccctta agtgggggat aacatttgga 120
aacagatgct aataccgcat agatccaaga accgcatggt tcttggctga aagatggcgt 180
aagctatcgc ttttggatgg acccgcggcg tattagctag ttggtgaggt aatggctcac 240
caaggcgatg atacgtagcc gaactgagag gttgatcggc cacattggga ctgagacacg 300
gcccaaactc ctacgggagg cagcagtagg gaatcttcca caatggacgc aagtctgatg 360
gagcaacgcc gcgtgagtga agaaggcttt cgggtcgtaa aactctgttg ttggagaaga 420
atggtcggca gagtaactgt tgtcggcgtg acggtatcca accagaaagc cacggctaac 480
tacgtgccag cagccgcggt aatacgtagg tggcaagcgt tatccggatt tattgggcgt 540
aaagcgagcg caggcggttt tttaagtctg atgtgaaagc cctcggctta accgaggaag 600
cgcatcggaa actgggaaac ttgagtgcag aagaggacag tggaactcca tgtgtagcgg 660
tgaaatgcgt agatatatgg aagaacacca gtggcgaagg cggctgtctg gtctgtaact 720
gacgctgagg ctcgaaagca tgggtagcga acaggattag ataccctggt agtccatgcc 780
gtaaacgatg aatgctaggt gttggagggt ttccgccctt cagtgccgca gctaacgcat 840
taagcattcc gcctggggag tacgaccgca aggttgaaac tcaaaggaat tgacgggggc 900
ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 960
ttgacatctt ttgatcacct gagagatcag gtttcccctt cgggggcaaa atgacaggtg 1020
gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080
acccttatga ctagttgcca gcatttagtt gggcactcta gtaagactgc cggtgacaaa 1140
ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg 1200
tgctacaatg gatggtacaa cgagttgcga gaccgcgagg tcaagctaat ctcttaaagc 1260
cattctcagt tcggactgta ggctgcaact cgcctacacg aagtcggaat cgctagtaat 1320
cgcggatcag cacgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac 1380
catgagagtt tgtaacaccc gaagccggtg gcgtaaccct ttaggagcga gcc 1433
Claims (7)
1.一株副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)BB4,于2018年9月10日保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为:CGMCC No.16455。
2.根据权利要求1所述的副干酪乳杆菌副干酪亚种,其特征在于,菌株的形态特征为:菌体呈杆状;在MRS平板培养基上培养24~48 h后,菌落呈圆形,乳白色,表面光滑、***,边缘整齐,不透明,菌体呈杆状,大小为0.3~0.6 × 1.0~3.0 μm。
3.一种如权利要求1所述的副干酪乳杆菌副干酪亚种在制备抑制创伤弧菌药物或普通食品中的应用。
4.根据权利要求3所述的应用,其特征在于,所述药物或普通食品中所含副干酪乳杆菌副干酪亚种BB4的活菌数为1.0×106-5.0×1011 cfu/mL或1.0×106-5.0×1011 cfu/g。
5.根据权利要求3所述的应用,其特征在于,所述药物或普通食品中所含副干酪乳杆菌副干酪亚种BB4的活菌数为2.0×106-2.0×108 cfu/mL或2.0×106-2.0×108 cfu/g。
6.根据权利要求3所述的应用,其特征在于,所述药物或普通食品中所含副干酪乳杆菌副干酪亚种BB4的活菌数为2.0×107 cfu/mL或2.0×107 cfu/g。
7.一种如权利要求1所述的副干酪乳杆菌副干酪亚种在降解偶氮类人工合成色素中的应用。
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