CN114847475B - 一种促进akk菌生长的发酵槐米粉的应用 - Google Patents
一种促进akk菌生长的发酵槐米粉的应用 Download PDFInfo
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Abstract
本发明公开了一种促进AKK菌生长的发酵槐米粉的应用，所述发酵槐米粉是槐米粉经发酵过的植物乳酸杆菌发酵而成；经过验证发现，AKK菌能显著降低血清的甘油三酯、谷丙转氨酶（ALT）、谷草转氨酶（AST）和碱性磷酸酶（ALP）的含量，并可观察到肝脏的脂肪沉积显著降低，还具有减肥的效果。所以该发酵槐米粉可用于制备减轻脂肪肝和肥胖症状的食品或药品，安全性高，无毒副作用，显效快。
Description
技术领域
本发明涉及食品原料领域，具体涉及一种促进AKK菌生长的发酵槐米粉的应用。
背景技术
脂肪肝是一种较长时间形成的疾病。脂肪肝危害巨大，需及早预防。形成脂肪肝的因素还需要进一步分析证明，有研究显示可能与长时间高血压、血糖异常、肥胖、饮食生活不良的习惯等因素有关。
脂肪肝是肝脏病变的早期临床表现，如果不及时治疗，将进一步演变为肝炎、肝纤维化、肝硬化甚至肝癌。临床研究中脂肪肝主要分为酒精性脂肪肝和非酒精性脂肪肝，酒精性脂肪肝的发病原因主要是乙醇的过量积累，导致酒精在肝脏内蓄积，后续发生脂质过氧化反应，导致脂肪肝。非酒精性脂肪肝的发病原因是除酒精以外的其他因素导致肝脏中三酰甘油的异常积累，导致发生脂肪肝。目前，临床中对于脂肪肝的治疗还没有研发出特效药，且药物治疗费用昂贵且副作用较多。
发明内容
为了克服现有技术的不足，本发明的目的在于提供一种促进AKK菌生长的发酵槐米粉的应用，该槐米粉能上调AKK菌的菌群含量，能减轻脂肪肝和肥胖的症状，而且安全性高，无毒副作用，显效快。
本发明的目的采用如下技术方案实现：
一种促进AKK菌生长的发酵槐米粉的应用，所述发酵槐米粉是槐米粉经发酵过的植物乳酸杆菌发酵而成；所述发酵槐米粉用于制备减轻脂肪肝和肥胖症状的食品或药品。优选地，所述发酵槐米粉的用于制备能减少腹部脂肪的食品或药品，即所述发酵槐米粉的减肥效果主要体现在减少腹部脂肪。
进一步，所述发酵过的植物乳酸杆菌的制备方法为：从酸奶中筛选得植物乳酸杆菌，将筛选得到的植物乳酸杆菌经液体培养基培养发酵，得到发酵过的植物乳酸杆菌。
再进一步，所述液体培养基包括以下组分：蛋白胨、牛肉膏、酵母粉、葡萄糖、磷酸氢二钾、乙酸钠、柠檬酸氢二铵、硫酸镁和硫酸锰。
进一步，所述发酵槐米粉的制备方法为：将所述发酵过的植物乳酸杆菌接种到发酵底物中，调节水分后进行发酵，得到发酵产物进行烘干粉粹，得到发酵槐米粉。
再进一步，所述发酵底物包括以下按重量百分比计的原料：槐米粉80~98%、环糊精2~20%，鼠李糖苷酶0.01~1%、磷酸氢二钾0.03~1%和硫酸镁0.01~0.5%。
进一步，所述发酵过的植物乳酸杆菌的接种量为0.5~3wt%。
再进一步，调节水分至30~60%，混合均匀后在25~35℃后发酵2~4天。
进一步，所述食品或药品的剂型为片剂、胶囊剂、颗粒剂、粉剂或口服液中的一种。
相比现有技术，本发明的有益效果在于：
（1）本发明的发酵槐米粉能提高AKK菌的菌种含量，经过验证发现，AKK菌能显著降低小鼠血清的甘油三酯、谷丙转氨酶（ALT）、谷草转氨酶（AST）和碱性磷酸酶（ALP）的含量，并可观察到肝脏的脂肪沉积显著降低；还具有减肥的效果。所以该发酵槐米粉可用于制备减轻脂肪肝和肥胖症状的食品或药品，而且安全性高，无毒副作用，显效快。
（2）本发明的发酵槐米粉的减肥作用进一步可以体现在减腹部脂肪上。
（3）本发明发酵槐米粉是槐米粉经发酵过的植物乳酸杆菌发酵而成，工艺简单，可用于大规模生产发酵槐米粉。
（4）本发明的发酵槐米粉可用于各种食品、药品场景，如饮料、糕点烘焙类、保健品及药用等领域。
附图说明
图1为各组肠道AKK菌的数量柱状图；
图2为各组小鼠ALP指标的柱状图；
图3为各组小鼠ALT指标的柱状图；
图4为各组小鼠AST指标的柱状图；
图5为各组小鼠的血液甘油三脂的柱状图；
图6为各组小鼠的肝脏组织切片示意图；
图7为各组小鼠的体重柱状图；
图8为各组小鼠的肠系膜脂肪重量柱状图。
具体实施方式
下面，结合附图以及具体实施方式，对本发明做进一步描述，需要说明的是，在不相冲突的前提下，以下描述的各实施例之间或各技术特征之间可以任意组合形成新的实施例。
实施例1
发酵槐米粉的制备方法，包括以下步骤：
1）菌种的制备：从植物乳杆菌的平板培养基中挑取一个单菌落，接中到液体培养基中，在30℃，200 rpm条件下震荡培养48 h，离心收集菌体并确定菌体湿重；
所述液体培养基包括以下组分：蛋白胨10.0 g/L，牛肉膏5.0 g/L，酵母粉5.0 g/L，葡萄糖20.0 g/L，磷酸氢二钾2.0 g/L，乙酸钠5.0 g/L，柠檬酸氢二铵2.0 g/L，硫酸镁0.2 g/L，硫酸锰0.054 g/L，pH 6.0，115℃下灭菌20 min。
2）发酵过程：将步骤1）获得的菌体以0.5wt%的接种量加入发酵底物中，调节水分至30%，充分混匀，在30℃下静置发酵2天；其中，所述发酵底物包括以下组分：槐米粉95%，环糊精4.5%，鼠李糖苷酶0. 1%，磷酸氢二钾0.3%，硫酸镁0.1%。
3）后处理将步骤2）发酵完成的槐米粉进行低温冷风烘干、粉碎并做一定的微生物灭菌处理，获得发酵槐米粉。
效果测试
1.小鼠的饲养及分组
雄性C57BL6小鼠（6-8周），并在23.2℃下进行12小时的光-暗循环。经过一周的适应后，雄性c57bl/6小鼠（20-25g体重）在屏障动物设施中驯化1w后，分别饲养在恒温（22±1℃）、光/暗循环（12h/12h）、湿度（40-60%）的特定无病原体室内。分别按照如下组进行喂养，期间小鼠被允许随意获得水和食物，小鼠以各自的饮食维持15周。所有小鼠随机分为4组（n=8）：（1）对照：标准正常饮食；（2）HFD：高脂肪饮食；（3）HFD/槐米提取物：高脂饮食+0.5%槐米提取物；（4）HFD/发酵槐米提取物：高脂饮食+0.5%发酵槐米提取物。其中，槐米提取物即为槐米粉，发酵槐米提取物即为上述发酵槐米粉。
2.实验观察、记录及检测
每周测量体重和食物摄入量，每日观察小鼠食欲、排便、尿液、活动等情况，每周记录小鼠体重，绘制小鼠体重变化曲线。15周后对小鼠称量体重、肠系膜脂肪重量并进行血清检测（观察血清中的碱性磷酸酶（ALP）、谷丙转氨酶（ALT）、谷草转氨酶（AST）、甘油三酯，检测方法按照试剂盒的说明书来完成），处死小鼠摘取肝脏，并制作组织切片（观察肝脏的组织切片HE染色和油红O染色），去肠内容物进行16s核糖体测序观察肠道AKK菌的数量变化。
3.实验结果
3.1肠道AKK菌的变化
由图1可得，HFD组的小鼠AKK菌的数量很大量的下调，HFD/槐米提取物组的AKK菌的数量有少许回调，但是回调数量不多，而HFD/发酵槐米提取物组小鼠的AKK菌数量较HFD组来说其菌群数量大量回调，也比HFD/槐米提取物组好。因此可以推断经过发酵的槐米有调节肠道AKK菌的作用。
3.2肝功能指标观察
由图2-4可以看出，HFD组的所有小鼠的肝功能指标（ALP、AST、ALT）较control组显著上升，可见HFD组的小鼠肝功能显著下降，槐米提取物组的肝功能有所恢复，但恢复的不够大，发酵槐米提取物组的肝功能显著恢复，且其水平接近control组。可见发酵过的槐米具有保护治疗肝损伤的作用，且效果显著。
3.3血液甘油三脂的指标观察
由图5结果可得，HFD组的血液甘油三脂的值显著增加，槐米提取物组的有所降低，但其降低的量还没有发酵槐米提取物组的高。可见发酵后的槐米降脂的作用较强。
3.4治疗脂肪肝的作用
由图6肝脏的组织切片可见，肝脏的切片HE染色发现HFD组的肝组织可见大量的白色脂肪样变细胞，HFD/槐米提取物组也同样有数量不少的白色脂肪样变细胞，反观HFD/发酵槐米提取物组的肝脏组织似乎很少脂肪样变细胞。
进一步进行脂肪染色观察脂肪（油红O染色），发现HFD组和HFD/槐米提取物组大部分被染成红色，说明这两组出现大量的脂肪滴，有大量的脂肪沉积，即形成脂肪肝，而HFD/发酵槐米提取物组几乎很少红色的细胞，这个结果和HE染色观察到的结果是一致的，可见发酵后的槐米具有强大的治疗脂肪肝的作用。
3.5体重观察
由图7可见，HFD组小鼠体重显著升高，发酵槐米提取物组体重相对于HFD组显著的降低。可见发酵后的槐米具有降体重的作用。图8的结果可见，HFD组小鼠肠系膜脂肪重量显著升高，发酵槐米提取物组体重相对于HFD组显著的降低，可见发酵槐米粉的减肥效果进一步体现在减腹部脂肪上。
综上所述，本发明提供的槐米粉具有上调肠道AKK菌的作用，能提高肝功能，降低血脂，保护肝脏以及减轻脂肪肝的作用，同时还能减肥，更突出的是具有减腹部脂肪的效果。上述实施方式仅为本发明的优选实施方式，不能以此来限定本发明保护的范围，本领域的技术人员在本发明的基础上所做的任何非实质性的变化及替换均属于本发明所要求保护范围。
Claims (3)
1.一种发酵槐米粉在制备促进肠道AKK菌生长的产品中的应用，其特征在于，所述发酵槐米粉是槐米粉经发酵过的植物乳酸杆菌发酵而成；所述发酵过的植物乳酸杆菌的制备方法为：从酸奶中筛选得植物乳酸杆菌，将筛选得到的植物乳酸杆菌经液体培养基培养发酵，得到发酵过的植物乳酸杆菌；所述液体培养基包括以下组分：蛋白胨、牛肉膏、酵母粉、葡萄糖、磷酸氢二钾、乙酸钠、柠檬酸氢二铵、硫酸镁和硫酸锰；所述发酵槐米粉的制备方法为：将所述发酵过的植物乳酸杆菌接种到发酵底物中，调节水分后进行发酵，得到发酵产物进行烘干粉粹，得到发酵槐米粉；其中，所述发酵底物包括以下组分：槐米粉95%，环糊精4.5%，鼠李糖苷酶0. 1%，磷酸氢二钾0.3%，硫酸镁0.1%。
2.如权利要求1所述的发酵槐米粉在制备促进肠道AKK菌生长的产品中的应用，其特征在于，所述发酵过的植物乳酸杆菌的接种量为0.5~3wt%。
3.如权利要求1所述的发酵槐米粉在制备促进肠道AKK菌生长的产品中的应用，其特征在于，调节水分至30~60%，混合均匀后在25~35℃发酵2~4天。
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