CN111705012B - 一种具有促进消化作用的副干酪乳杆菌lc-37及应用 - Google Patents
一种具有促进消化作用的副干酪乳杆菌lc-37及应用 Download PDFInfo
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Abstract
本发明属于微生物发酵领域,具体涉及一种具有促进消化作用的副干酪乳杆菌(Lactobacillus paracasei)LC‑37及应用,提供的一种副干酪乳杆菌(Lactobacillus paracasei)LC‑37,已保藏于中国微生物菌种保藏管理委员会普通微生物中心,其保藏编号是CGMCC NO.14055。所述的副干酪乳杆菌LC‑37分离自广西巴马健康长寿老人肠道内容物,所述的副干酪乳杆菌LC‑37具有促进消化的作用,具体表现为对于消化不良人群腹痛、嗳气、泛酸、腹胀、食欲不振、腹泻或便秘等消化不良症状的改善,同时可以增加肠道中短链脂肪酸的含量。
Description
技术领域
本发明属于微生物发酵领域,具体涉及一种具有促进消化作用的副干酪乳杆菌(Lactobacillus paracasei)LC-37及应用。
背景技术
副干酪乳杆菌(Lactobacillus paracasei)是乳杆菌属(Lactobacillus) 中的种群之一,是重要的益生菌,常应用于乳制品、保健食品、饮料和冰淇淋等生产。已有众多研究表明副干酪乳杆菌具有调节肠道菌群、增强免疫力等功能。如中国专利文献CN110892989A中公开的一种具有缓解便秘功能的副干酪乳杆菌ET-22,其中介绍的是一种具有缓解便秘功能的副干酪乳杆菌ET-22,该菌株最主要的功能为提高肠道推进率,缩短肠道运转时间,从而起到缓解便秘的作用。再如中国专利文献CN110882280A中公开的一种副干酪乳杆菌K56的新应用,其中介绍的是副干酪乳杆菌CGMCC15139显著缓解和/或预防便秘的功效,能够提高肠道推进率、缩短排便时间。
随着社会的发展,生活节奏不断加快,工作压力变大,不能保证按时进餐、饮食快餐化等问题造成的消化不良问题逐渐凸显。食用具有促进消化作用的食品是改善消化不良问题的重要而便捷的手段。然而,目前对于副干酪乳杆菌的促进消化作用的研究较少,而且应用于乳酸菌饮料产品开发的益生菌主要依赖于进口,缺少我们的自主知识产权的菌株,并且缺少促进消化相关功能的有效评价。
发明内容
因此,本发明要解决的技术问题在于提供一种具有促进消化作用的副干酪乳杆菌(Lactobacillus paracasei)LC-37及应用。
本发明提供了一种副干酪乳杆菌(Lactobacillus paracasei)菌株LC-37,已保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC),保藏地址: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,邮政编码:100101,其保藏编号是CGMCCNO.14055,保藏日期是2017 年4月20日。
本发明提供了一种微生物制剂,所述微生物制剂是以所述副干酪乳杆菌LC-37、所述副干酪乳杆菌LC-37的灭活物或所述副干酪乳杆菌LC-37 的处理物为活性成分的微生物制剂;
所述副干酪乳杆菌LC-37的处理物为副干酪乳杆菌LC-37的培养物、破碎物和所述培养物的提取物中的至少一种。
所述的微生物制剂,还包括至少一种适合于微生物制剂的赋形剂;所述微生物制剂为粉末、丸剂、胶囊、颗粒剂、片剂、液体制剂或者凝胶剂。
本发明提供了一种副干酪乳杆菌LC-37菌粉的制备方法,由副干酪乳杆菌LC-37的发酵液离心,收集菌体,向所得菌体中加入冻干保护剂,真空冷冻干燥,得到冻干粉。
在所述的制备方法中,副干酪乳杆菌LC-37的发酵步骤如下:将副干酪乳杆菌LC-37的种子液以2%~5%(v/v)的接种量接种至发酵培养基中,在33~40℃下培养,搅拌转速100~200rpm,培养基保持pH 5.0~6.5,发酵 14~20小时。
在所述的制备方法中,离心条件为:转速为8000~10000rpm,离心1~2 小时。
在所述的制备方法中,冻干保护剂包括如下重量份的原料:海藻糖1~3 份,脱脂乳粉1~3份、麦芽糊精2~4份和抗坏血酸钠0.1~0.5份。
冻干保护剂加入量与离心所得菌体的质量比为0.2~0.9:1,真空冷冻干燥的条件为40-60小时内温度为-50℃升至30℃,真空度为3~50Pa。
在所述的制备方法中,还包括种子培养液的制备步骤:将副干酪乳杆菌LC-37冻存菌种接种至10~50mL培养基中,放置于33~40℃培养12~24 小时。
在所述的制备方法中,还包括向所述冻干粉中加入辅料,辅料可以是麦芽糊精、乳清粉、低聚果糖或抗性淀粉等。
本发明提供了一种由上述的副干酪乳杆菌LC-37菌粉的制备方法制备得到的副干酪乳杆菌LC-37菌粉。
本发明提供了一种所述的副干酪乳杆菌LC-37、所述的微生物制剂、所述的方法制备得到的菌粉或所述的菌粉在制备具有促进消化作用的药物或食品中的用途。
本发明中,药物可以根据给药途径不同而制备成不同形式,例如,可制备成散剂、片剂、颗粒剂、胶囊剂、溶液剂、乳剂、混悬剂等形式;药物中还可包括药学上可接受的佐剂。食品包括任意类型的食品,例如固体饮料、豆制品、果汁、乳制品、冰淇淋、糖果、饼干等;食品中还可含有常规的添加剂、营养强化剂、原辅料,例如香精香料、稳定剂、增稠剂、防腐剂、矿物质、维生素、麦芽糊精等。
本发明提供了一种含副干酪乳杆菌LC-37的乳酸菌饮品的制备方法,包括使用所述的副干酪乳杆菌LC-37、所述的微生物制剂、所述的方法制备得到的菌粉或所述的菌粉。
在上述的制备方法中,在饮品基料的发酵步骤中接种副干酪乳杆菌 LC-37菌粉,接种量为104-107CFU/mL,发酵温度为33-40℃,发酵时间为 72-100小时。
在上述的制备方法中,制备得到的乳酸菌饮品中副干酪乳杆菌LC-37 的活菌数范围为≥5×107CFU/mL。
在上述的制备方法中,所述饮品基料包括如下重量份的原料:脱脂乳粉110-140份,葡萄糖15-25份,和白砂糖35-50份。
在上述的制备方法中,所述饮品基料在发酵前经过杀菌褐变,杀菌褐变条件为90-97℃,2-5小时。
在上述的制备方法中,还包括在发酵步骤获得的发酵乳中加入辅料。所述辅料可以为糖液和/或香精等。
本发明提供了一种如所述的方法制备得到的含副干酪乳杆菌LC-37的乳酸菌饮品。
本发明相对现有技术具有如下优点:
1.本发明提供的一种副干酪乳杆菌(Lactobacillus paracasei)LC-37,已保藏于中国微生物菌种保藏管理委员会普通微生物中心,其保藏编号是 CGMCC NO.14055。所述的副干酪乳杆菌LC-37分离自广西巴马健康长寿老人肠道内容物,所述的副干酪乳杆菌LC-37具有改善消化不良的作用,具体表现为对于消化不良人群腹痛、嗳气、泛酸、腹胀、食欲不振、腹泻或便秘等消化不良症状的改善,同时可以增加肠道中短链脂肪酸的含量。
2.本发明提供的含副干酪乳杆菌LC-37的乳酸菌饮品,利用副干酪乳杆菌菌株LC-37作为发酵菌种,制备得到的乳酸菌饮品具有促进消化的作用,能够改善腹痛、嗳气、泛酸、腹胀、食欲不振、腹泻或便秘等消化不良症状,同时可以增加肠道中短链脂肪酸的含量。
3.本发明提供的副干酪乳杆菌LC-37菌粉,具有促进消化的作用,能够改善腹痛、嗳气、泛酸、腹胀、食欲不振、腹泻或便秘等消化不良症状,同时可以增加肠道中短链脂肪酸的含量。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案,下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图是本发明的一些实施方式,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1是本发明实验例中受试者的腹痛症状的评分变化;
图2是本发明实验例中受试者的嗳气症状的评分变化;
图3是本发明实验例中受试者的泛酸症状的评分变化;
图4是本发明实验例中受试者的腹胀症状的评分变化;
图5是本发明实验例中受试者的食欲症状的评分变化;
图6是本发明实验例中受试者的腹泻及腹泻症状的评分变化;
图7是本发明实验例中受试者的消化不良总临床症状积分的变化;
图8是本发明实验例中受试者饮用前后乙酸含量的变化;
图9是本发明实验例中受试者饮用前后丙酸含量的变化;
图10是本发明实验例中受试者饮用前后丁酸含量的变化;
图11是本发明实验例中受试者饮用前后总酸含量的变化;
图12是本发明实验例中标准曲线图。
具体实施方式
提供下述实施例是为了更好地进一步理解本发明,并不局限于所述最佳实施方式,不对本发明的内容和保护范围构成限制,任何人在本发明的启示下或是将本发明与其他现有技术的特征进行组合而得出的任何与本发明相同或相近似的产品,均落在本发明的保护范围之内。
实施例中未注明具体实验步骤或条件者,按照本领域内的文献所描述的常规实验步骤的操作或条件即可进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规试剂产品。
以下通过具体实施例来说明本发明的实施方式,除非另外说明,本发明中所公开的实验方法均采用本技术领域常规技术,实施例中所用到的试剂和原料均可由市场购得。
实施例1副干酪乳杆菌菌株LC-37的获得
1.副干酪乳杆菌菌株LC-37的分离
采集广西巴马健康长寿老人肠道内容物,也即粪便,使用稀释液对广西巴马健康长寿老人肠道粪便样品进行梯度稀释,使用MRS培养基(乳酸细菌培养基)分离乳杆菌,挑取不同形态菌落,镜检选取革兰氏阳性的杆菌,继续使用倾注平板法纯化菌株,直到分离得到菌株的纯培养物。经过上述分离过程,得到一株乳杆菌。
上述稀释步骤中使用的稀释液的配制方法:磷酸二氢钠4.5g,磷酸氢二钠6.0g,L-半胱氨酸0.5g,琼脂0.5g,吐温80为0.5mL,蒸馏水1000 mL,加热溶解,调pH至7.4-7.6,121℃下灭菌15min,备用。
所述倾注平板法为:挑取不同形态菌落接种到液体MRS培养基中培养 12-16h,取1mL菌液使用稀释液连续进行7次10倍梯度稀释,取1mL稀释后菌液加入无菌平皿中,倒入适量MRS固体培养基,待培养基凝固后,倒置放入37℃培养箱中培养48h,挑取单菌落获得菌株的纯培养物。
2.鉴定和保藏
将上述分离得到的一株乳杆菌经16S rRNA鉴定为副干酪乳杆菌,命名为副干酪乳杆菌LC-37,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,邮政编码:100101,保藏编号为CGMCCNo.14055,保藏日期是 2017年4月20日。
实施例2副干酪乳杆菌LC-37菌粉
本实施例提供一种副干酪乳杆菌LC-37菌粉的制备方法,具体包括以下步骤:
1.菌种的扩大培养
将副干酪乳杆菌LC-37冻存菌种接种至10mL MRS培养基中,放置于 37℃培养12小时,得到适于接种的种子培养液,镜检菌株形态与菌株特有形态相符,无可见杂菌。
2.发酵
将上述得到的种子培养液以2%~5%(v/v)(在本实施例中选择2%v/v) 的接种量接种至发酵培养基中,在本实施例中发酵培养基为MRS培养基在33℃下培养,搅拌转速200rpm,培养基pH保持pH 5.0,发酵20小时。发酵结束后使用碟片式离心机对培养液进行离心,离心机转速为8000rpm,离心2小时,收集菌体。
3.冻干
向收集的菌体中加入冻干保护剂,在本实施例中冻干保护剂包括如下重量份的原料:海藻糖1kg,脱脂乳粉3kg、麦芽糊精2kg、抗坏血酸钠0.5kg。冻干保护剂加入量与离心所得菌体的质量比例为0.9:1,放入冷冻干燥盘中,真空冷冻干燥条件为40小时内温度为-50℃升至30℃,真空度为3Pa,使最终菌粉的水分活度低于0.25,真空冷冻干燥结束。随后进行粉碎和包装,形成副干酪乳杆菌LC-37冻干粉,根据副干酪乳杆菌LC-37冻干粉的活菌数,按比例混入麦芽糊精,使得最终得到副干酪乳杆菌LC-37菌粉中的活菌数为5×109CFU/g。
实施例3副干酪乳杆菌LC-37菌粉
本实施例提供一种副干酪乳杆菌LC-37菌粉的制备方法,具体包括以下步骤:
1.菌种的扩大培养
将副干酪乳杆菌LC-37冻存菌种接种至10mL MRS培养基中,放置于 37℃培养12小时,得到适于接种的种子培养液,镜检菌株形态与菌株特有形态相符,无可见杂菌。
2.发酵
将上述得到的种子培养液以2%~5%(v/v)(在本实施例中选择5%v/v) 的接种量接种至发酵培养基中,在本实施例中发酵培养基为MRS培养基,在40℃下培养,搅拌转速100rpm,培养基pH保持pH6.0,发酵14小时。发酵结束后使用碟片式离心机对培养液进行离心,离心机转速为10000rpm,离心1小时,收集菌体。
3.冻干
向收集的菌体中加入冻干保护剂,在本实施例中冻干保护剂包括如下重量份的原料:海藻糖3kg,脱脂乳粉1kg、麦芽糊精4kg、抗坏血酸钠0.1kg,冻干保护剂加入量与离心所得菌体的比例为0.5:1,放入冷冻干燥盘中,真空冷冻干燥条为60小时内温度为-50℃升至30℃,真空度为50Pa,使最终菌粉的水分活度低于0.25,真空冷冻干燥结束。随后进行粉碎和包装,形成副干酪乳杆菌LC-37冻干粉,根据副干酪乳杆菌LC-37冻干粉的活菌数,按比例混入麦芽糊精,使得最终得到副干酪乳杆菌LC-37菌粉中活菌数为5 ×1010CFU/g。
实施例4副干酪乳杆菌LC-37菌粉
本实施例提供一种副干酪乳杆菌LC-37菌粉的制备方法,具体包括以下步骤:
1.菌种的扩大培养
将副干酪乳杆菌LC-37冻存菌种接种至10mL MRS培养基中,放置于 37℃培养12小时,得到适于接种的种子培养液,镜检菌株形态与菌株特有形态相符,无可见杂菌。
2.发酵
将上述得到的种子培养液以2%~5%(v/v)(在本实施例中选择3%v/v) 的接种量接种至发酵培养基中,在本实施例中发酵培养基为MRS培养基,在37℃下培养,搅拌转速150rpm,培养基pH保持pH 5.8,发酵17小时。发酵结束后使用碟片式离心机对培养液进行离心,离心机转速为9000rpm,离心1.5小时,收集菌体。
3.冻干
向收集的菌体中加入冻干保护剂,在本实施例中冻干保护剂包括如下重量份的原料:海藻糖2kg,脱脂乳粉2kg、麦芽糊精3kg、抗坏血酸钠0.3kg。冻干保护剂加入量与离心所得菌体的比例为0.2:1,放入冷冻干燥盘中,真空冷冻干燥条件为50小时内温度为-50℃升至30℃,真空度为25Pa,使最终菌粉的水分活度低于0.25,真空冷冻干燥结束。随后进行粉碎和包装,形成副干酪乳杆菌LC-37冻干粉,根据副干酪乳杆菌LC-37冻干粉的活菌数,按比例混入麦芽糊精,使得最终得到副干酪乳杆菌LC-37菌粉中的活菌数为5×1011CFU/g。
实施例5含副干酪乳杆菌LC-37的乳酸菌饮品
本实施例提供了一种含副干酪乳杆菌LC-37的乳酸菌饮品的制备方法,包括如下步骤:
以脱脂乳粉(110kg)为原料,添加葡萄糖(25kg)、白砂糖(35kg),经杀菌、褐变,即于90℃下杀菌、褐变5小时,随后冷却至33℃,接种实施例2中制备的副干酪乳杆菌LC-37菌粉,接种量为107CFU/mL(即使原料液中副干酪乳杆菌LC-37的终浓度为107CFU/mL),发酵温度为33℃,经长时间发酵72小时,结束发酵,然后再以所得的发酵乳为主要原料,添加适量糖液、香精等辅料,使得最终制成含副干酪乳杆菌LC-37的乳酸菌饮料中活菌数含量:5×107CFU/ml。
实施例6含副干酪乳杆菌LC-37的乳酸菌饮品
本实施例提供了一种含副干酪乳杆菌LC-37的乳酸菌饮品的制备方法,包括如下步骤:
以脱脂乳粉(140kg)为原料,添加葡萄糖(15kg)、白砂糖(50kg),经杀菌、褐变,即于97℃下杀菌、褐变2小时,随后冷却至40℃,接种实施例3中制备的副干酪乳杆菌LC-37菌粉,接种量为104CFU/mL(即使原料液中副干酪乳杆菌LC-37的终浓度为104CFU/mL),发酵温度为40℃,经长时间发酵100小时,结束发酵,然后再以所得的发酵乳为主要原料,添加适量糖液、香精等辅料,使得最终制成含副干酪乳杆菌LC-37的乳酸菌饮料中活菌数含量:1×108CFU/ml。
实施例7含副干酪乳杆菌LC-37的乳酸菌饮品
本实施例提供了一种含副干酪乳杆菌LC-37的乳酸菌饮品的制备方法,包括如下步骤:
以脱脂乳粉(125kg)为原料,添加葡萄糖(20kg)、白砂糖(42kg),经杀菌、褐变,即于90-97℃下杀菌、褐变2-5小时,随后冷却至33-40℃,接种实施例4中制备的副干酪乳杆菌LC-37菌粉,接种量为105CFU/mL(使即原料液中副干酪乳杆菌LC-37的终浓度为105CFU/mL),发酵温度为37℃,经长时间发酵86小时,结束发酵,然后再以所得的发酵乳为主要原料,添加适量糖液、香精等辅料,使得最终制成含副干酪乳杆菌LC-37的乳酸菌饮料中的活菌数含量:5×108CFU/ml。
实验例
1.受试对象
1.1纳入者标准
选择功能性消化不良,伴有长期胃肠不适,主诉食欲不振,早饱、气多,胃肠胀满,呕吐,不明原因慢性腹泻或大便秘结等自愿受试者。
1.2排除者标准
(1)急性腹泻者。
(2)严重器质性病变引起的消化不良者。
(3)体质虚弱无法接受试验者。
(4)合并有心血管、肝、肾和造血系统等严重全身性疾病患者。
(5)未按要求服用受试样品,无法判断饮用结果者。
(6)乳糖不耐受或牛奶过敏者。
1.3试验设计
实验经过筛选,入组人群共计120人,分为4组,每组30人。其中男性55人,女性65人;年龄20-60岁,平均年龄为47.8岁,平均BMI指数为24.4。BMI指数:身体质量指数(BodyMass Index,简称BMI)。
1.4食用剂量及时间
样品置于2-10℃冰箱存放,每天午饭或晚饭后服用,A、C组每天服用副干酪乳杆菌LC-37菌粉一袋,B、D组每天服用一瓶含有副干酪乳杆菌 LC-37的乳酸菌饮品。连续服用28天。试验期间不改变原来的饮食习惯,正常饮食。
A组:副干酪乳杆菌LC-37菌粉1(实施例2制备的活菌数含量:5× 109CFU/g,每天摄入量:2g);
B组:含有副干酪乳杆菌LC-37的乳酸菌饮料1(实施例5制备的活菌数含量:5×107CFU/ml,每天摄入量:200ml)
C组:副干酪乳杆菌LC-37菌粉2(实施例3制备的活菌数含量:5× 1010CFU/g,每天摄入量:2g);
D组:含有副干酪乳杆菌LC-37的乳酸菌饮料2(实施例7制备的活菌数含量:5×108CFU/ml,每天摄入量:200ml)
1.5实验流程
排空期共持续7天。期间,志愿者不可饮用任何发酵制品(包括酸奶、奶酪、活性乳酸菌饮料等),但可饮用牛奶。第7天为第一次粪便样品和血液采集时间,并做临床症状评分、填写肠道健康调查。饮用期共持续28天,第14天第二次采集粪便,做临床症状评分,填写健康调查表。第28天采集粪便样品和血液,做临床症状评分、填写肠道健康调查表。
1.6观察指标
1.6.1安全性指标
对饮用前后所采集的血液样本进行血常规、血生化指标的检测。各干预组血液样本中白细胞计数、血红蛋白、平均红细胞容积、平均红细胞血红蛋白浓度、血小板计数、血小板压积、淋巴细胞绝对值、中性粒细胞绝对值、嗜碱性粒细胞绝对值、单核细胞百分数、嗜酸性粒细胞百分数、红细胞计数、红细胞比容、平均红细胞血红蛋白量、红细胞体积分布宽度、单核细胞绝对值、嗜酸性粒细胞绝对值、淋巴细胞百分数、中性粒细胞百分数、嗜碱性粒细胞百分数、尿素、尿酸、谷草转氨酶、总蛋白、白球比、甘油三酯、肌酐、谷丙转氨酶、白蛋白、球蛋白、总胆固醇、血糖等血常规、血生化指标均在正常值范围内,说明副干酪乳杆菌LC-37及含有副干酪乳杆菌LC-37的乳酸菌饮料对人体均无不良影响。具体见下表1-表4:
表1 A组饮用前各组血常规和血生化
表2 B组饮用前各组血常规和血生化
表3 C组饮用前各组血常规和血生化
表4 D组饮用前各组血常规和血生化
1.6.2功效性观察
准确记录受试者试验前后消化不良的临床症状,按表5给予量化评分,比较试验前后消化不良症状积分的变化。
表5临床症状积分
将腹痛、嗳气、泛酸、腹胀、食欲、腹泻或便秘的症状评分相加获得消化不良临床症状积分。对人群的临床症状积分进行统计分析,结果如图 1-图7所示。图1-图6为各项消化不良症状的评分变化,图7为消化不良总临床症状积分的变化。由图1-图7可以看到,第0天时,各组人群腹痛、嗳气、泛酸等各项消化不良症状评分及消化不良症状总积分均相近,无显著差异,说明各组人群具有可比性。在干预期14天时,各组的消化不良评分均出现了显著的降低。而且,相比较而言,高剂量副干酪乳杆菌的效果要优于低剂量副干酪乳杆菌,含有同剂量副干酪乳杆菌的乳酸菌饮料优于副干酪乳杆菌菌粉。在干预期第28天时,各组的消化不良症状评分进一步降低,其中所有组别的食欲不振症状消失,D组腹痛、嗳气、泛酸症状消失。各组消化不良总症状积分由最初的4.18~4.71降低至0.17~0.71,出现了极显著的改善。
1.6.3粪便中短链脂肪酸含量检测
短链脂肪酸标准曲线的建立:用容量瓶配制含有5mmol/L乙酸、5 mmol/L丙酸、5mmol/L丁酸、5mmol/L庚酸的乙醚溶液,采用倍比稀释的方法,得到分别含有2.5,1.25,0.625,0.312,0.156mmol/L的乙酸、丙酸、丁酸、庚酸的标准溶液。分别移取1μL标准液作气相色谱分析,每个浓度重复测定3次,得到各浓度的峰面积与对应浓度,从而制作标准曲线,并求得回归方程和相关系数。
粪便中短链脂肪酸的提取:在1.5mL螺口收集管内加入适量玻璃珠 (2~3mm),加入1mL含有庚酸(浓度为1mmol/L)的乙醚溶液和50μL 的盐酸溶液(浓度为1mmol/L),称重。用镊子向收集管中小心加入受试者粪便样品(约0.1-0.2g),再称次重,记录加入粪便的重量。配平后用均质机匀浆1min,10000r/min离心2min。转移上清液有内插管的进样瓶中。因乙醚具有强挥发性,上述操作均需在冰浴或低温条件下快速进行。
检测粪便样品中短链脂肪酸浓度的色谱条件:检测器为FID氢火焰离子检测器;色谱柱DB-FFAP色谱柱;升温程序:50℃保持1min,以10℃ /min升至140℃,保持1min,以30℃/min升至240℃,保持2min;载气 (N2)压力260kPa;氮气流速为40mL/min,氢气流速为40mL/min,空气流速为400mL/min;进样量2μL;检测温度230℃。N2000色谱工作站收集信号并检测。
按照上述气相色谱条件进行检测,计算对应短链脂肪酸的峰面积,采用外标法做出标准曲线(如图12所示),用内标庚酸浓度进行修正,计算出检测样品中乙酸、丙酸和丁酸浓度,并换算出其在粪便样品中的含量。
检测结果如图8-11所示,在食用副干酪乳杆菌LC-37菌粉及含有副干酪乳杆菌LC-37的乳酸菌饮料14天后,受试人群粪便中乙酸、丙酸、丁酸及总短链脂肪酸的含量无显著变化。在食用28天后,各组乙酸、丙酸、丁酸及总短链脂肪酸均出现了不同程度的上升。表明食用副干酪乳杆菌LC-37 菌粉和含有副干酪乳杆菌LC-37的乳酸菌饮料能够增加粪便中短链脂肪酸含量。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (11)
1.一种副干酪乳杆菌(Lactobacillus paracasei)LC-37,已保藏于中国微生物菌种保藏管理委员会普通微生物中心,其保藏编号是CGMCC NO.14055。
2.一种微生物制剂,其特征在于,所述微生物制剂是以权利要求1所述的副干酪乳杆菌LC-37为活性成分的微生物制剂。
3.根据权利要求 2 所述的微生物制剂,其特征在于,还包括至少一种适合于微生物制剂的赋形剂;所述微生物制剂为粉末、丸剂、胶囊、颗粒剂、片剂、液体制剂或者凝胶剂。
4.一种副干酪乳杆菌LC-37菌粉的制备方法,其特征在于,由权利要求1所述的副干酪乳杆菌LC-37的发酵液离心,收集菌体,向所得菌体中加入冻干保护剂,真空冷冻干燥,得到冻干粉。
5.一种由权利要求4所述的制备方法制备得到的副干酪乳杆菌LC-37菌粉。
6.权利要求1所述的副干酪乳杆菌LC-37、权利要求2或3所述的微生物制剂、权利要求4所述的方法制备得到的菌粉或权利要求5所述的菌粉在制备具有促进消化作用的药物中的用途。
7.权利要求1所述的副干酪乳杆菌LC-37、权利要求2或3所述的微生物制剂、权利要求4所述的方法制备得到的菌粉或权利要求5所述的菌粉在制备具有促进消化作用的食品中的用途。
8.一种含副干酪乳杆菌LC-37的乳酸菌饮品的制备方法,其特征在于,包括使用权利要求1所述的副干酪乳杆菌LC-37、权利要求2或3所述的微生物制剂、权利要求4所述的方法制备得到的菌粉或权利要求5所述的菌粉。
9.根据权利要求8所述的制备方法,其特征在于,包括如下步骤:
在饮品基料的发酵步骤中接种副干酪乳杆菌LC-37菌粉,接种量为104-107 CFU/mL,发酵温度为33-40℃,发酵时间为72-100小时,制备得到的乳酸菌饮品中副干酪乳杆菌LC-37的活菌数范围为≥5×107CFU/mL。
10.根据权利要求9所述的方法,其特征在于,所述饮品基料包括如下重量份的原料:脱脂乳粉110-140份,葡萄糖15-25份,和白砂糖35-50份;所述饮品基料在发酵前经过杀菌褐变,杀菌褐变条件为90-97℃,2-5小时。
11.一种如权利要求8-10任一项所述的方法制备得到的含副干酪乳杆菌LC-37的乳酸菌饮品。
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