CN109957530B - 一种植物乳杆菌及其应用 - Google Patents
一种植物乳杆菌及其应用 Download PDFInfo
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- CN109957530B CN109957530B CN201910207822.XA CN201910207822A CN109957530B CN 109957530 B CN109957530 B CN 109957530B CN 201910207822 A CN201910207822 A CN 201910207822A CN 109957530 B CN109957530 B CN 109957530B
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- lactobacillus plantarum
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Abstract
本发明公开了一种植物乳杆菌及其应用，属于微生物技术领域，该菌种命名为植物乳杆菌（Lactobacillus plantarum）HCS03‑001。该菌株已于2018年08月14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”，保藏号为CGMCC No.16258。本发明筛选的植物乳杆菌抑口腔致病菌能力强、具有通便和抗腹泻的功效。
Description
技术领域
本发明属于生物技术领域，特别涉及一株抑口腔致病菌能力强、具有通便和抗腹泻功效的植物乳杆菌及其应用。
本发明所述的所述菌株命名为植物乳杆菌（Lactobacillus plantarum）HCS03-001，该菌株已于2018年08月14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”，保藏号为CGMCC No.16258。
背景技术
当内在或外在因素变化造成口腔中的微生态系统失去平衡时，就易引起常见的口腔疾病，如龋齿、牙周炎、牙龋炎和口臭等。龋齿和牙周病是一种发病率极高的口腔疾病，主要由变异链球菌和牙龈卟啉单胞菌等口腔致病菌引起的。由于抗生素治疗口腔疾病易使口腔致病菌产生抗药性，所以利用具有抑菌能力并可以提高机体抵抗力的益生菌治疗口腔疾病已逐渐成为研究的热点。
便秘是临床常见的复杂症状，主要是指排便次数减少、粪便量减少、粪便干结、排便费力等，若不及时缓解会影响身体健康。腹泻是由多病原、多因素引起的以大便次数增多和大便性状改变为特点的消化道综合征，可出现脱水、电解质紊乱，严重者可有昏迷、惊厥等。益生菌是双重调节剂，具有缓解便秘和防治腹泻的功效。
植物乳杆菌是乳酸菌的一种，2010年被列入卫生部办公厅公布的《可用于食品的菌种名单》。植物乳杆菌能够通过其代谢过程中形成的乳酸杆菌素、有机酸、过氧化氢等多种天然抑菌物质对其他细菌作用，达到抑制病原菌的生长、调节肠道内菌群平衡、促进矿物质的吸收、降低胆固醇水平、缓解乳糖不耐症、增强免疫力及抑制肿瘤细胞的形成等生理功效。但目前几乎没有关于植物乳杆菌具有抑制口腔致病菌（牙龈卟啉单胞菌和变异链球菌）能力的相关报道。
发明内容
本发明目的是提供一株抑口腔致病菌能力强、具有通便和抗腹泻功效的植物乳杆菌。
本发明所述的植物乳杆菌，所述菌株命名为植物乳杆菌（Lactobacillus plantarum）HCS03-001，该菌株已于2018年08月14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”，保藏号为CGMCC No.16258。
上述植物乳杆菌分离自顺产健康8月龄的女婴粪便中，是一株抑口腔致病菌能力强、具有通便和抗腹泻功效的植物乳杆菌，具有植物乳杆菌的典型特征，即：革兰氏阳性，兼性厌氧，无芽孢，不运动；菌落较小且大小均一，边缘整齐；细胞呈短杆状，成对或成链排列；其理化特征是：接触酶阴性，氧化酶阴性，上述植物乳杆菌CGMCC No.16258的细胞形态和理化实验结果详见表1。
表1 植物乳杆菌CGMCC No.16258的细胞形态和理化实验结果
上述植物乳杆菌CGMCC No.16258生长温度为36~38℃，最适生长温度为37℃。
上述植物乳杆菌CGMCC No.16258在MRS改良培养基中37℃静置培养17小时，pH值降至3.91，检测活菌数为4.0×109cfu/mL。
上述MRS改良培养基的配方：蛋白胨 10g/L，牛肉粉 3g/L，酵母粉 4g/L，磷酸氢二钾 2g/L，柠檬酸三铵 2g/L，乙酸钠 5g/L，葡萄糖 20g/L，七水合硫酸镁 0.58g/L，四水合硫酸锰 0.15g/L，吐温-80 0.6g/L，调pH至6.5；115℃，30min灭菌。
通过在美国生物工程信息中心（National Center for BiotechnologyInformation，NCBI）查阅国际相关的基因库，将本发明的植物乳杆菌CGMCC No.16258 16SrDNA基因序列与有关菌株16S rDNA基因序列比较，虽有相似性，但却不完全相同，说明本发明的菌株是首次分离鉴定，具有原创性。
表2 植物乳杆菌CGMCC No.16258的16S rDNA序列与GenBank提交菌株
序列相似性比较
将上述植物乳杆菌制备成发酵用的菌种冻存管，冻存的菌种复苏后，经过液体培养基一级、二级、三级培养，以及菌种四级发酵得到植物乳杆菌的发酵液，检测其活菌数大于109cfu/mL，发酵液经过离心，离心后的菌泥添加冻干保护剂，经过真空冷冻干燥制成植物乳杆菌冻干粉，检测其活菌数可达5.0×1011 cfu/g以上，存活率为90%。
本发明公开了一株抑口腔致病菌能力强、具有通便和抗腹泻功效的的植物乳杆菌，且该菌在通过含有胃酸和胆盐的消化道后依然保持较高活性，为其发挥功能特性提供了基础条件。
上述植物乳杆菌（Lactobacillus plantarum）HCS03-001具有以下优点：
1.本发明的植物乳杆菌（Lactobacillus plantarum）HCS03-001分离筛选自健康顺产8月龄女婴粪便中，确保其来源的安全性，而且更利于定植在婴幼儿的肠道中。通常益生菌可以更好的在与其来源相似的环境中发挥功效。
2.本发明的植物乳杆菌（Lactobacillus plantarum）HCS03-001对缓解便秘和防治腹泻有双重功效，尤其是对于那些由于胃肠道正常菌群紊乱引起的便秘和腹泻。
3.本发明的植物乳杆菌（Lactobacillus plantarum）HCS03-001具有较强的耐胃液、耐肠液的能力，可通过胃肠道的逆环境有效定植于肠道内，发挥其功能特性。
4.牙龈卟啉单胞菌是一种非酵解糖的革兰氏阴性厌氧球杆菌，属于重要的牙周致病菌，是引起牙周病的一种主要细菌。变异链球菌是口腔主要的致龋齿菌之一，常见于牙菌斑中，可通过代谢产生的酸性物质导致龋齿的发生。本发明的植物乳杆菌（Lactobacillus plantarum）HCS03-001具有较好的抑菌特性，尤其对口腔主要致病菌（牙龈卟啉单胞菌和变异链球菌）具有较强的抑制作用。
5.本发明的植物乳杆菌（Lactobacillus plantarum）HCS03-001制成冻干粉后活菌数可达5千亿cfu/g以上，存活率高，定植能力强。
本发明所述的抑口腔致病菌能力强、具有通便和抗腹泻功效的植物乳杆菌（Lactobacillus plantarum）HCS03-001，其已于2018年08月14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”（地址：北京市朝阳区北辰西路1号院3号），保藏号为CGMCC No.16258。
具体实施方式
实施例1植物乳杆菌（Lactobacillus plantarum）HCS03-001 CGMCC No.16258菌株的筛选及生理生化特性
取健康顺产8月龄女婴粪便5g加到5mL缓冲蛋白胨液中，10倍系列稀释，选择合适稀释度，分别划线到MRS改良培养基平板上。37℃培养24~48h（平板放入自封袋中）；挑取具有目的菌株典型特征、菌落较大、活性较强的单菌落，于MRS改良培养基进行划线纯化培养，如此反复2~3次，直到划线平皿中菌落特征一致；每个纯化后的平皿挑取2个以上单菌落进行涂片、革兰氏染色，并且在显微镜下观察其颜色、菌体形状是否一致，从而确定该平皿中菌落是否为纯培养物。如镜下观察结果一致，则将得到的纯培养物（平皿菌落）作为疑似菌株，待鉴定；如镜下观察结果不一致，则继续进行上述操作。最终得到的疑似菌株进行糖发酵鉴定、API鉴定、16s rDNA全序列测序鉴定。最终筛选得到一株植物乳杆菌，命名为植物乳杆菌（Lactobacillus plantarum）HCS03-001，已于2018年08月14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”，保藏号为CGMCC No.16258。
上述植物乳杆菌（Lactobacillus plantarum）HCS03-001菌株为革兰氏阳性菌，兼性厌氧，无芽孢，不运动；菌落较小且大小均一，边缘整齐，表面光滑细密，白色或乳白色；菌体呈短杆状，成对或成链排列；其生理生化特征是：接触酶阴性，氧化酶阴性，可利用L-阿拉伯糖、D-核糖、D-半乳糖、D-葡萄糖、D-果糖、D-甘露糖、甘露醇、山梨醇、α-甲基-D-甘露糖甙、苦杏仁甙、熊果甙、七叶灵、水杨苷、纤维二糖、麦芽糖、乳糖、蜜二糖、蔗糖、海藻糖、松三糖、棉子糖、D-松二糖。
实施例2筛选的植物乳杆菌HCS03-001耐消化道逆环境实验
2.1 模拟胃液试验
将传代三次的菌液震荡摇匀，取菌悬液10mL离心（5000×g，10min，5℃）获得菌泥，用PBS缓冲液冲洗3次，获得的菌泥重悬于10 mL模拟胃液中，37℃条件下消化3h，取样测活菌数。
活化三代后的受试菌株在第三代培养液中的活菌数用N′表示，经在模拟胃液培养基中消化3h后计数测得的活菌数用N″表示，受试菌株模拟胃液试验存活率计算公式如下：
受试菌株模拟胃液试验存活率（%）=lg cfu N″/lg cfu N′ ×100%；
表3 HCS03-001模拟胃液试验数据表
HCS03-001菌株在模拟胃液中的存活能力较强，经3h处理，活菌对数值略有下降趋势，仍可达到8.6以上。
2.2模拟肠液试验
将传代三次的菌液震荡摇匀，取菌悬液10mL离心（5000×g，10min，5℃）获得菌泥，用PBS缓冲液冲洗2次，获得的菌泥重悬于10 mL的人工肠液中，37℃条件下培养，于2h、4h分别取样测活菌数。
活化三代后的受试菌株在第三代培养液中的活菌数用N′表示，经在模拟肠液培养液中培养2h、4h后计数测得的活菌数用N″表示，受试菌株模拟肠液试验存活率计算公式如下：
受试菌株模拟肠液试验存活率（%）=lg cfu N″/lg cfu N′ ×100%；
表4HCS03-001模拟肠液试验数据表
HCS03-001菌株在模拟肠液中的存存活能力较强，随处理时间延长，活菌对数值基本没有下降，4h后可达到9.4。
综上，说明该菌株能够有效抵抗胃肠液的影响，从而能够在通过消化道后依然维持较高活性。
实施例3菌株抑菌功能特性实验
指示菌株准备：取指示菌株冻存管在37℃水浴锅中使其迅速溶化，用微量移液器取100μL菌液加入10ml液体培养基中，大肠杆菌/金黄色葡萄球菌/鼠伤寒沙门氏菌/志贺氏菌/单增李斯特菌37℃恒温过夜培养；白色念珠菌28℃恒温摇床培养48h，转速100 rpm ；牙龈卟啉单胞菌厌氧37℃恒温过夜培养；变异链球菌厌氧30℃恒温过夜培养。
制备固体培养基，将培养至对数期的致病菌稀释至菌液浓度为105cfu/mL，取菌悬液100μL均匀涂布于固体培养基上，用镊子将无菌牛津杯竖于涂布有指示菌的平皿中，每个牛津杯中加入200μL受试样品。将平皿置于适宜温度的恒温培养箱中，培养适当时间后观察，并用游标卡尺计量抑菌圈直径。
实验结果见表5。表5中采用牛津杯法，牛津杯内直径6mm，外直径8mm；使用受试菌株发酵液上清，分别进行3倍、5倍浓缩以及直接取原上清进行试验。
表5抑菌谱记录表
通过试验结果可以得出结论，HCS03-001对胃肠道中主要致病菌有明显抑菌作用，尤其对口腔主要致病菌（牙龈卟啉单胞菌和变异链球菌）具有较强的抑制作用；对包括革兰氏阳性细菌和革兰氏阴性细菌，均有较强的抑制作用，但对于真菌（白色念珠菌）没有抑菌效果。
实施例4植物乳杆菌（Lactobacillus plantarum）HCS03-001 CGMCC No.16258制成植物乳杆菌冻干粉
包括如下步骤：
一、发酵用菌种的制备
1.1）菌种划线培养：植物乳杆菌菌粉用无菌水稀释后，于MC培养基上四区划线，划线结束后，平皿倒置，37℃培养箱恒温培养60小时；
1.2）一级纯化培养：挑取平皿上溶钙圈较大的单菌落接种至装有5ml MRS液体培养基的试管中，试管密封，37℃培养箱恒温静置培养15小时；
1.3）二级纯化培养：按3%接种量，将一级纯化培养好的菌悬液接种至装有100mlMRS液体培养基的250ml三角瓶中，三角瓶密封，37℃培养箱恒温静置培养20小时；
1.4）菌种保存：对二级纯化培养得到的100ml菌悬液进行离心，12000rpm离心10分钟后得到菌泥，将菌泥溶解于10ml混合液（混合液由5ml MRS液体培养基与5ml 50%甘油溶液混合均匀配制而成）中，得菌泥混合液，将菌泥混合液分装在已灭菌的1ml冻存管中，封口膜密封，冻存于-80℃低温冰箱内，得植物乳杆菌菌泥冻存管，备用；
注：1.1）—1.4）操作均必需在无菌操作台和无菌间内执行；
二、菌种的发酵
2.1）冻存菌种复苏：取存于低温冰箱内的植物乳杆菌菌泥冻存管，立即放入37℃水浴锅内进行菌种复苏，30～45s，至冻存管内液体全部融化；
2.2）菌种一级培养：将复苏好的1ml菌泥混合液直接接种至装有10ml基础培养基的50ml三角瓶中，三角瓶密封，37℃培养箱恒温静置培养20小时；
2.3）菌种二级培养：按照2%接种量，将一级培养结束的菌悬液接种至装有100ml基础培养基的250ml三角瓶中，按此接种4瓶，三角瓶密封，37℃培养箱恒温静置培养20小时；
2.4）菌种三级培养：按照5%接种量，将二级培养结束的菌悬液接种至装有7.2L基础培养基的12L发酵罐中，开启发酵罐搅拌桨，转速为100rpm，通气量为0，37℃恒温培养10小时；
2.5）菌种四级发酵：按照5%接种量，将三级培养结束的菌悬液接种至装有120L优化培养基的200L发酵罐中，开启发酵罐搅拌桨，转速为100rpm，通气量为0，37℃恒温培养，在发酵开始3小时后，通过流加食品级 NaOH溶液来维持恒定pH为5.10±0.1，培养至10小时时，监测到菌液OD600值停止增长，立即停止发酵；
其中，基础培养基的配方为：酵母蛋白胨 8.0 g/L、葡萄糖 25.0 g/L、酵母浸出物4.0 g/L、乙酸钠5.0 g/L、硫酸镁 0.3 g/L、吐温80 0.5 g/L、磷酸二氢钾 3.0 g/L、余量为无菌水，pH值6.20。制备：按照配方比例称量、加热溶解，用1mol/L食品级 NaOH溶液调培养基pH值至6.20， 115℃下灭菌30min；
优化培养基配方：酵母蛋白胨 18.0 g/L、葡萄糖 32.0 g/L、酵母浸出物 12.0 g/L、乙酸钠 10.0 g/L、硫酸镁 0.1 g/L、吐温80 0.5 g/L、磷酸二氢钾 3.0 g/L、余量为无菌水，pH值6.20。制备：按照配方比例称量、加热溶解，用1mol/L食品级 NaOH溶液调培养基pH值至6.20， 115℃下灭菌30min；
注：培养基中各组分原料均为食品级；
三、冻干粉的制备
3.1）发酵液离心：四级发酵结束后，将发酵罐罐体温度调低，当罐内发酵液温度低于20℃时准备离心，离心设备采用管式离心机，离心前对转鼓进行蒸汽灭菌30min，离心转速为12000rpm；
3.2）菌泥收集，离心结束后，收集到转鼓内的菌泥2.03kg，菌泥收率为1.69%；
3.3）冻干保护剂添加，菌泥收集完毕后，按照菌泥：冻干保护剂=1:2～3的质量体积比，添加冻干保护剂，并搅拌、得到混合均匀的菌粉。
按照冻干保护剂的配方配制5L冻干保护剂。
其冻干保护剂的配方：脱脂奶粉 75.0 g/L、麦芽糊精 25.0 g/L、海藻糖 150.0g/L、吐温80 0.6 g/L、甘油 10.0 g/L、谷氨酸钠1.2 g/L、抗坏血酸钠1.2 g/L、余量为无菌水。制备：按照比例对各组分进行称量、溶解，于115℃灭菌30min，待温度降至室温后，冷藏于-4℃冰箱内备用。
3.4）冷冻干燥：将混合均匀的菌粉，分装到冻干机托盘中，然后将冻干机托盘放入冻干机内进行菌粉的冻干，真空度0～1.0Pa。冷冻干燥机的温度和冻干时间按区段设置，具体为，第1区段：冻干时长2h，温度设定为-40℃；第2区段：冻干时长7h，温度设定为-20℃；第3区段：冻干时长12h，温度设定为-10℃；第4区段：冻干时长17h，温度设定为10℃；第5区段：冻干时长10h，温度设定为20℃；第6区段：冻干时长2h，温度设定为30℃。
3.5）冻干粉收集，检测活菌数为5.85×1011 cfu/g。
3.6）粉碎、包装，按质量要求进行粉碎后包装。
注：2.1—3.6）操作均需在10万级净化车间内进行。
实施例5植物乳杆菌（Lactobacillus plantarum）HCS03-001 CGMCC No.16258通便功效实验
5.1小肠运动实验
实验动物：4～5周龄清洁级昆明小鼠，18～22g，雄性，共50只。
实验内容：经口灌胃给予造模药物盐酸洛哌丁胺，建立小鼠小肠便秘模型，计算一定时间内小肠的墨汁推进率，来判断小肠蠕动功能。试验设置3个剂量组，一个溶剂对照组和一个模型对照组。受试物剂量组为0.75g/kg、1.5g/kg、3.0g/kg，每天灌胃一次，灌胃体积为10ml/kg，共给予21天。
判定标准：单位时间推进率明显高于模型对照组，可判定该项结果为阳性。
表6植物乳杆菌（Lactobacillus plantarum）HCS03-001通便功效（小肠运动实验）试验结果
用植物乳杆菌（HCS03-001）作为受试物进行了小鼠通便功效小肠运动试验。由表6结果可知，植物乳杆菌（HCS03-001）在0.75g/kg、1.5g/kg剂量与3.0g/kg剂量下,单位时间推进率明显高于模型对照组，对盐酸洛哌丁胺造模的便秘模型均有一定的通便效果，结果为阳性，且随添加剂量的增加通便效果更佳。
5.2排便粒数、排便时间实验
实验动物：4～5周龄清洁级昆明小鼠，18～22g，雄性，共40只。
实验内容：经口灌胃给予造模药物盐酸洛哌丁胺，建立小鼠小肠便秘模型，考察实验动物6h内排黑便粒数与首次排黑便时间，来反映模型排便情况。试验设置2个剂量组，一个溶剂对照组和一个模型对照组。受试物剂量组为1.5g/kg、3.0g/kg，每天灌胃一次，灌胃体积为10ml/kg，共给予8天。
判定标准：首粒排黑便时间明显短于模型对照组，排便时间项目结果可判定为阳性；
排便粒数明显高于模型对照组，排便粒数项目结果可判定为阳性。
表7植物乳杆菌（Lactobacillus plantarum）HCS03-001通便功效
（排便粒数、排便时间）试验结果
*p﹤0.05与模型对照组比较
用植物乳杆菌（HCS03-001）作为受试物进行了小鼠通便功效排便粒数、排便时间试验。由表7可知，植物乳杆菌（HCS03-001）在3.0g/kg剂量下，首粒排黑便时间明显短于模型对照组，排便粒数明显高于模型对照组，具有较好的通便功效。
综上，植物乳杆菌（HCS03-001）作为受试物进行了小鼠通便功效试验，小肠运动实验与排便粒数、排便时间实验的实验结果均为阳性，即可判断植物乳杆菌（HCS03-001）具有较好的通便功效。
实施例6植物乳杆菌（Lactobacillus plantarum）HCS03-001 CGMCC No.16258抗腹泻功效实验
实验动物：4～5周龄清洁级昆明小鼠，18～22g，雄性，共30只。
实验内容：经口灌胃给予造模药物番泻叶，建立小鼠腹泻模型，考察实验动物6h内腹泻次数，来反映模型排便情况。试验设置2个剂量组和一个模型对照组。受试物剂量组为1.5g/kg、3.0g/kg，每天灌胃一次，灌胃体积为10ml/kg，共给予7天。
表8 植物乳杆菌（HCS03-001）抗腹泻功效试验结果
*p﹤0.05与模型对照组比较
用植物乳杆菌（HCS03-001）作为受试物进行了小鼠抗腹泻功效试验。由表8可知，植物乳杆菌（HCS03-001）在3.0g/kg剂量下，腹泻次数明显少于模型对照组，具有较好的抗腹泻功效。
<110> 汉臣氏（沈阳）儿童制品有限公司
<120>一种植物乳杆菌及其应用
<160> 1
<170> PatentIn Version 2.1
<210> 1
<211> 1408
<212> DNA
<213>植物乳杆菌（Lactobacillus plantarum）HCS03-001
<400> 1
ACCCCACCGA CTTTGGGTGT TACAAACTCT CATGGTGTGA CGGGCGGTGT GTACAAGGCC 60
CGGGAACGTA TTCACCGCGG CATGCTGATC CGCGATTACT AGCGATTCCG ACTTCATGTA 120
GGCGAGTTGC AGCCTACAAT CCGAACTGAG AATGGCTTTA AGAGATTAGC TTACTCTCGC 180
GAGTTCGCAA CTCGTTGTAC CATCCATTGT AGCACGTGTG TAGCCCAGGT CATAAGGGGC 240
ATGATGATTT GACGTCATCC CCACCTTCCT CCGGTTTGTC ACCGGCAGTC TCACCAGAGT 300
GCCCAACTTA ATGCTGGCAA CTGATAATAA GGGTTGCGCT CGTTGCGGGA CTTAACCCAA 360
CATCTCACGA CACGAGCTGA CGACAACCAT GCACCACCTG TATCCATGTC CCCGAAGGGA 420
ACGTCTAATC TCTTAGATTT GCATAGTATG TCAAGACCTG GTAAGGTTCT TCGCGTAGCT 480
TCGAATTAAA CCACATGCTC CACCGCTTGT GCGGGCCCCC GTCAATTCCT TTGAGTTTCA 540
GCCTTGCGGC CGTACTCCCC AGGCGGAATG CTTAATGCGT TAGCTGCAGC ACTGAAGGGC 600
GGAAACCCTC CAACACTTAG CATTCATCGT TTACGGTATG GACTACCAGG GTATCTAATC 660
CTGTTTGCTA CCCATACTTT CGAGCCTCAG CGTCAGTTAC AGACCAGACA GCCGCCTTCG 720
CCACTGGTGT TCTTCCATAT ATCTACGCAT TTCACCGCTA CACATGGAGT TCCACTGTCC 780
TCTTCTGCAC TCAAGTTTCC CAGTTTCCGA TGCACTTCTT CGGTTGAGCC GAAGGCTTTC 840
ACATCAGACT TAAAAAACCG CCTGCGCTCG CTTTACGCCC AATAAATCCG GACAACGCTT 900
GCCACCTACG TATTACCGCG GCTGCTGGCA CGTAGTTAGC CGTGGCTTTC TGGTTAAATA 960
CCGTCAATAC CTGAACAGTT ACTCTCAGAT ATGTTCTTCT TTAACAACAG AGTTTTACGA 1020
GCCGAAACCC TTCTTCACTC ACGCGGCGTT GCTCCATCAG ACTTTCGTCC ATTGTGGAAG 1080
ATTCCCTACT GCTGCCTCCC GTAGGAGTTT GGGCCGTGTC TCAGTCCCAA TGTGGCCGAT 1140
TACCCTCTCA GGTCGGCTAC GTATCATTGC CATGGTGAGC CGTTACCCCA CCATCTAGCT 1200
AATACGCCGC GGGACCATCC AAAAGTGATA GCCGAAGCCA TCTTTCAAGC TCGGACCATG 1260
CGGTCCAAGT TGTTATGCGG TATTAGCATC TGTTTCCAGG TGTTATCCCC CGCTTCTGGG 1320
CAGGTTTCCC ACGTGTTACT CACCAGTTCG CCACTCACTC AAATGTAAAT CATGATGCAA 1380
GCACCAATCA ATACCAGAGT TCGTTCGA 1408
Claims (2)
1.一种植物乳杆菌，其特征在于，所述的植物乳杆菌命名为植物乳杆菌（Lactobacillus plantarum）HCS03-001，该菌株已于2018年08月14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”，保藏号为CGMCC No.16258。
2.权利要求1所述的一种植物乳杆菌在制备具有抑制口腔致病菌、通便、抗腹泻功效的植物乳杆菌冻干粉中的应用。
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