CN108113001B - 一种缓解胃溃疡的益生菌冻干粉 - Google Patents
一种缓解胃溃疡的益生菌冻干粉 Download PDFInfo
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Abstract
本发明公开了一种治疗胃溃疡的益生菌冻干粉，其是以鼠李糖乳杆菌、乳双岐杆菌、干酪乳杆菌、嗜酸乳杆菌为益生菌菌种，将其经活化、扩大培养后，接种入由莼菜汁、山药粉、刺玫果粉、黑糖为原料制得的培养基中进行发酵，再经冷冻干燥制得。本发明所得益生菌冻干粉不仅具有健胃消食的功效，还对幽门螺旋杆菌引起的胃溃疡有明显疗效。
Description
技术领域
本发明属于食品技术领域，具体涉及一种缓解胃溃疡的益生菌冻干粉。
背景技术
现有研究表明，胃溃疡的发生与幽门螺旋杆菌有密切关系。胃幽门螺旋杆菌可黏附于胃黏膜上皮细胞，而避免随食物被胃排出；幽门螺旋杆菌分泌的氧化氢酶、蛋白水解酶及毒素等均会损伤胃黏膜上皮细胞；此外，幽门螺旋杆菌还可透过尿素酶水解尿素产生氨，在菌体周围形成“氨云”保护层，以抵抗胃酸的杀菌作用。
目前对幽门螺旋杆菌的治疗主要是采用抗生素，但抗生素对幽门螺旋杆菌的清除效果有限，且其在杀灭有害细菌的同时，也会抑制或杀死胃肠道内的一些有益菌。益生菌是一类对宿主有益的活性微生物，其定植于人体肠道、生殖***内，具有维护宿主胃肠道内菌群生态平衡、减少病菌毒素对胃肠道的损伤等多种有益效果。因此，采用益生菌剂治疗幽门螺旋杆菌引起的胃溃疡是目前研究的热点。
发明内容
本发明的目的在于提供一种缓解胃溃疡的益生菌冻干粉，其不仅具有健胃消食的功效，还对幽门螺旋杆菌引起的胃溃疡有明显疗效。
为实现上述目的，本发明采用如下技术方案：
一种缓解胃溃疡的益生菌冻干粉，其制备方法包括以下步骤：
1）益生菌的活化：将鼠李糖乳杆菌、乳双岐杆菌、干酪乳杆菌、嗜酸乳杆菌四种菌种分别接种到四个MRS斜面培养基上，于30-37℃培养10-16h，得到活化的斜面菌种；
2）扩大培养：将所得斜面菌种分别接种于MRS液体培养基中，30-37℃培养10-16h，得到液体菌种；然后将四种液体菌种分别按0.2-0.5%(v/v)的接种量接入灭菌的培养液中，30-37℃培养1-2d，得菌种扩大液；
3）发酵：以莼菜汁、山药粉、刺玫果粉、黑糖为原料制备发酵培养基，将其灭菌后按3-5%(v/v)的接种量接种菌种扩大液，30-37℃密封培养1-2d，至pH降至3.5-4.0时终止发酵；
4）冷冻干燥：将发酵产物过滤，所得发酵液经冷冻干燥制得所述益生菌冻干粉。
步骤2）所述培养液中含葡萄糖20g/L、蛋白胨8g/L、酵母膏6g/L、苜蓿提取物2g/L；
所述苜蓿提取物是在苜蓿中加入其重量3-5倍的水，100℃煎煮50-80min后过滤，然后在滤液中加入其体积60-80%的无水乙醇，静置8-10h后取沉淀烘干制得。
步骤3）发酵培养基中所用原料按重量份计包括：莼菜汁50-80份、山药粉20-30份、刺玫果粉10-25份、黑糖10-15份。
所得益生菌冻干粉中活菌数不低于6×109cfu/g。
本发明以鼠李糖乳杆菌、乳双岐杆菌、干酪乳杆菌、嗜酸乳杆菌为益生菌菌种，使所得益生菌冻干粉具有良好健胃消食的功效；同时，本发明在菌种扩大培养的培养液中加入苜蓿提取物，并以莼菜汁、山药粉、刺玫果粉、黑糖为原料制备用于菌种发酵的发酵培养基，可结合利用苜蓿、莼菜、山药、刺玫果等的活性成分，在提高益生菌活性的同时使各种活性成分充分融合、转化，以提高其对幽门螺旋杆菌的抑制效果，从而提高所得益生菌冻干粉对胃溃疡的治疗作用。其中，苜蓿提取物的使用可在一定程度上抑制胃酸的分泌，从而减少胃酸对胃黏膜的刺激；莼菜汁、山药粉的加入有助于减少幽门螺旋杆菌在胃黏膜上皮细胞的粘附；刺玫果粉的使用可进一步减轻幽门螺旋杆菌产生的毒素等对胃黏膜的刺激；而黑糖的使用有利于促进溃疡面的愈合。
具体实施方式
为了使本发明所述的内容更加便于理解，下面结合具体实施方式对本发明所述的技术方案做进一步的说明，但是本发明不仅限于此。
所用苜蓿提取物是在苜蓿中加入其重量3倍的水，100℃煎煮60min后过滤，然后在滤液中加入其体积60%的无水乙醇，静置10h后取沉淀烘干制得。
所用莼菜汁是将莼菜用水漂洗后，于100℃水中漂烫杀青6s，然后将杀青后的莼菜搅打成浆，再加入其重量10倍的水混合后，经胶体磨处理2次制得。
实施例1
1）益生菌的活化：将鼠李糖乳杆菌、乳双岐杆菌、干酪乳杆菌、嗜酸乳杆菌四种菌种分别接种到四个MRS斜面培养基上，于30℃培养16h，得到活化的斜面菌种；
2）扩大培养：将所得斜面菌种分别接种于MRS液体培养基中，30℃培养16h，得到液体菌种；然后将四种液体菌种分别按0.2%(v/v)的接种量接入灭菌的培养液中，30℃培养2d，得菌种扩大液；所述培养液中含葡萄糖20g/L、蛋白胨8g/L、酵母膏6g/L、苜蓿提取物2g/L；
3）发酵：以莼菜汁1.0kg、山药粉0.6kg、刺玫果粉0.5kg、黑糖0.3kg制备发酵培养基，将其灭菌后按3%(v/v)的接种量接种菌种扩大液，30℃密封培养2d，至pH降至3.5-4.0时终止发酵；
4）冷冻干燥：将发酵产物过滤，所得发酵液经冷冻干燥制得益生菌冻干粉。
实施例2
1）益生菌的活化：将鼠李糖乳杆菌、乳双岐杆菌、干酪乳杆菌、嗜酸乳杆菌四种菌种分别接种到四个MRS斜面培养基上，于35℃培养12h，得到活化的斜面菌种；
2）扩大培养：将所得斜面菌种分别接种于MRS液体培养基中，35℃培养12h，得到液体菌种；然后将四种液体菌种分别按0.3%(v/v)的接种量接入灭菌的培养液中，35℃培养2d，得菌种扩大液；所述培养液中含葡萄糖20g/L、蛋白胨8g/L、酵母膏6g/L、苜蓿提取物2g/L；
3）发酵：以莼菜汁1.5kg、山药粉0.5kg、刺玫果粉0.2kg、黑糖0.25kg制备发酵培养基，将其灭菌后按4%(v/v)的接种量接种菌种扩大液，35℃密封培养2d，至pH降至3.5-4.0时终止发酵；
4）冷冻干燥：将发酵产物过滤，所得发酵液经冷冻干燥制得益生菌冻干粉。
实施例3
1）益生菌的活化：将鼠李糖乳杆菌、乳双岐杆菌、干酪乳杆菌、嗜酸乳杆菌四种菌种分别接种到四个MRS斜面培养基上，于37℃培养10h，得到活化的斜面菌种；
2）扩大培养：将所得斜面菌种分别接种于MRS液体培养基中，37℃培养10h，得到液体菌种；然后将四种液体菌种分别按0.5%(v/v)的接种量接入灭菌的培养液中，37℃培养1d，得菌种扩大液；所述培养液中含葡萄糖20g/L、蛋白胨8g/L、酵母膏6g/L、苜蓿提取物2g/L；
3）发酵：以莼菜汁1.6kg、山药粉0.4kg、刺玫果粉0.3kg、黑糖0.2kg制备发酵培养基，将其灭菌后按5%(v/v)的接种量接种菌种扩大液，37℃密封培养1d，至pH降至3.5-4.0时终止发酵；
4）冷冻干燥：将发酵产物过滤，所得发酵液经冷冻干燥制得益生菌冻干粉。
对比例1
实施例3步骤2）培养液中含葡萄糖20g/L、蛋白胨8g/L、酵母膏6g/L，其余步骤按实施例3进行制备。
对比例2
实施例3步骤3）发酵培养基配方为水1.6kg、山药粉0.4kg、刺玫果粉0.3kg、黑糖0.2kg，其余步骤按实施例3进行制备。
对比例3
实施例3步骤3）发酵培养基配方为莼菜汁1.7kg、山药粉0.6kg、黑糖0.2kg，其余步骤按实施例3进行制备。
对比例4
实施例3步骤3）发酵培养基配方为莼菜汁1.6kg、山药粉0.4kg、刺玫果粉0.3kg、葡萄糖0.2kg，其余步骤按实施例3进行制备。
取试验大鼠60只，雌雄各半，禁食12h后，腹腔注射3%戊巴比妥钠进行麻醉，用酒精棉签消毒皮肤后剖开腹腔，将于冰醋酸中浸透的圆形纸片（直径5mm）贴于胃部前壁窦体交界处浆膜面上，重复3次，关腹，缝合创口。术后常规喂养3天，于第4天将动物随机分为6组，试验组每天灌胃给予2mL实施例1所得益生菌冻干粉的水溶液（20mg/mL），对比组每天分别灌胃给予2mL对比例1-4所得益生菌冻干粉的水溶液（20mg/mL），空白组每天灌胃给予等量清水，连续10天，末次给药12h后脱颈椎处死，剖腹取胃，用生理盐水洗涤胃内容物，然后将胃固定于10%甲醛中，15min后将胃平展于玻璃表面皿上，量取溃疡面积，其结果见表1。
表1 不同处理对大鼠胃溃疡的影响
由表1可见，与空白组及对比组相比，本发明益生菌冻干粉可明显抑制胃黏膜损伤大鼠的溃疡指数。
以上所述仅为本发明的较佳实施例，凡依本发明申请专利范围所做的均等变化与修饰，皆应属本发明的涵盖范围。
Claims (1)
1.一种缓解胃溃疡的益生菌冻干粉的制备方法，其特征在于，包括以下步骤：
1）益生菌的活化：将鼠李糖乳杆菌、乳双歧杆菌、干酪乳杆菌、嗜酸乳杆菌四种菌种分别接种到四个MRS斜面培养基上，于30-37℃培养10-16h，得到活化的斜面菌种；
2）扩大培养：将所得斜面菌种分别接种于MRS液体培养基中，30-37℃培养10-16h，得到液体菌种；然后将四种液体菌种分别按0.2-0.5%v/v的接种量接入灭菌的培养液中，30-37℃培养1-2d，得菌种扩大液；
3）发酵：以莼菜汁、山药粉、刺玫果粉、黑糖为原料制备发酵培养基，将其灭菌后按3-5%v/v的接种量接种菌种扩大液，30-37℃密封培养1-2d，至pH降至3.5-4.0时终止发酵；
4）冷冻干燥：将发酵产物过滤，所得发酵液经冷冻干燥制得所述益生菌冻干粉；
步骤2）所述培养液中含葡萄糖20g/L、蛋白胨8g/L、酵母膏6g/L、苜蓿提取物2g/L；
所述苜蓿提取物是在苜蓿中加入其重量3-5倍的水，100℃煎煮50-80min后过滤，然后在滤液中加入其体积60-80%的无水乙醇，静置8-10h后取沉淀烘干制得；
步骤3）发酵培养基中所用原料按重量份计包括：莼菜汁50-80份、山药粉20-30份、刺玫果粉10-25份、黑糖10-15份。
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