CN110025014A - 混合菌株发酵制备富含益生菌和nk细胞的基底液的方法 - Google Patents
混合菌株发酵制备富含益生菌和nk细胞的基底液的方法 Download PDFInfo
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Abstract
本发明公开了一种混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，包括以下步骤：（1）菌株混合：选取八种菌株混合，加入甘油，放在室内细胞培养皿，得混合菌株；加入冷冻保护剂，低温保存于菌株库；（2）发酵：选取特选糖类作为发酵媒介（其在发酵过程中可完全消耗），将混合菌株和发酵媒介倒入烧瓶内，摇匀培养；转移至生物反应器，连续培养；最终转化为高生物活性乳酸菌；（3）NK细胞培养：通过五阶段生物发酵得NK细胞培养。有益效果：本发明制备工艺制得的肌底液帮助人体快速修复受损细胞，提高人体免疫机能，增强有益细胞活性；调节肠道益生菌，增强人体NK细胞的活性，使细胞活化，提高新陈代谢。
Description
技术领域
本发明属于保健品领域，具体涉及一种混合菌株发酵制备富含益生菌和NK细胞的基底液的方法。
背景技术
为了防止病原体的侵入和恶性肿瘤的发生等对机体不利的病态，机体通过天然免疫和适应性免疫这2类免疫体系来应对。天然免疫中，对于侵入的病原体，已经存在的NK细胞和巨噬细胞等吞噬细胞或者补体系统等通过非特异性活化通路在数小时内感知并迅速将其排除。适应性免疫中，对于特定的病原体或异常细胞，抗原特异性的效应T细胞被诱导并伴有增殖，变成具有细胞杀伤活性的T细胞，并且促进与抗原相对应的抗体生成，需要4天以上的时间来排除侵入的病原体。
在天然免疫中起到重要作用的NK细胞占外周血淋巴细胞的10％左右，进行免疫学监视，即迅速察觉机体的异常并进行应对。NK细胞在形态学上是具有特征性细胞内颗粒的大型的淋巴细胞，是无需像细胞杀伤性T细胞那样经过抗原刺激后的分化就可发挥细胞杀伤活性的细胞。因此，不具有像T细胞那样的抗原特异性，对主要组织相容性复合体(majorhistocompatibility complex，以下称为MHC)消失了的某种病毒感染细胞或癌细胞显示细胞杀伤活性。另一方面，NK细胞存在与像正常细胞那样表达有MHC-Ⅰ类分子的细胞结合的抑制性受体，因此对于正常细胞不发挥细胞杀伤活性。
除了非特异性的细胞杀伤活性之外，NK细胞还可通过特有的机制杀伤靶细胞。大多数NK细胞在细胞表面上表达有针对IgG分子的Fc部分的受体(CD16)。NK细胞介以CD16与结合于靶细胞的细胞表面的IgG的Fc部分结合而活化，发挥杀伤靶细胞的抗体依赖性细胞介导的细胞毒作用(ADCC)活性。
益生菌具有调整肠道菌群，改善 人体微生态平衡，抑制有害的病原微生物在人体中的生长繁殖等功效。目前常见的益生菌有乳酸菌，双歧杆菌，保加利亚乳杆菌，嗜热链球菌等。 注意，乳酸菌包括很多种菌属，双歧和保加利亚和嗜熱都是乳酸菌。
目前市售的益生菌食用产品大多是由单一菌种的菌粉和辅料经过复配后加工而成，菌粉经过多次加工后，菌种活力下降，不足以达到预期的保健效果。
发明内容
针对现有技术中的不足之处，本发明提供一种混合菌株发酵制备富含益生菌和NK细胞的基底液的方法。
为了达到上述目的，本发明技术方案如下：
混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，包括以下步骤：
（1）菌株混合：选取八种菌株混合，加入甘油，放在室内细胞培养皿，得混合菌株；加入冷冻保护剂，低温保存于菌株库；
（2）发酵：选取特选糖类作为发酵媒介（其在发酵过程中可完全消耗），将混合菌株和发酵媒介倒入烧瓶内，摇匀培养；转移至生物反应器，连续培养；最终转化为高生物活性乳酸菌；
（3）NK细胞培养：通过五阶段生物发酵得NK细胞培养。
进一步的，步骤（2）中生物反应器的容积为5L。
进一步的，步骤（2）中连续培养从5L到1000L生物反应器，称重与分配，喂养培养基与接种物。
进一步的，八种菌株为嗜酸乳杆菌、干酪乳杆菌、植物乳杆菌、保加利亚乳酸杆菌、发酵乳杆菌、乳酸双歧杆菌、两歧双歧杆菌和酵母菌。
进一步的，发酵媒介还可选取番木瓜果实及叶子、生糖、RO纯净水的混合物。
有益效果：本发明制备工艺制得的肌底液帮助人体快速修复受损细胞，提高人体免疫机能，增强有益细胞活性；调节肠道益生菌，增强人体NK细胞的活性，使细胞活化，提高新陈代谢。
附图说明
图1为本发明的制备工艺流程框图。
具体实施方式
以下参照具体的实施例来说明本发明。本领域技术人员能够理解，这些实施例仅用于说明本发明，其不以任何方式限制本发明的范围。
请参阅图1，混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，包括以下步骤：
（1）菌株混合：选取八种菌株混合，再加入甘油，放在室内细胞培养皿，得混合菌株，备用；
（2）发酵：选取糖作为发酵媒介（其在发酵过程中可完全消耗），将混合菌株和发酵媒介倒入烧瓶内，摇匀培养；转移至生物反应器，连续培养；最终转化为高生物活性乳酸菌；
（3）NK细胞培养：通过五阶段生物发酵得NK细胞培养。
其中，步骤（2）中生物反应器的容积为5L。
其中，步骤（2）中连续培养从5L到1000L生物反应器，称重与分配，喂养培养基与接种物。
其中，八种菌株为嗜酸乳杆菌、干酪乳杆菌、植物乳杆菌、保加利亚乳酸杆菌、发酵乳杆菌、乳酸双歧杆菌、两歧双歧杆菌和酵母菌。
其中，发酵媒介还可选取番木瓜果实及叶子、生糖、RO纯净水的混合物。
Claims (5)
1.混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，其特征在于，包括以下步骤：
（1）菌株混合：选取八种菌株混合，加入甘油，放在室内细胞培养皿，得混合菌株；加入冷冻保护剂，低温保存于菌株库；
（2）发酵：选取特选糖类作为发酵媒介（其在发酵过程中可完全消耗），将混合菌株和发酵媒介倒入烧瓶内，摇匀培养；转移至生物反应器，连续培养；最终转化为高生物活性乳酸菌；
（3）NK细胞培养：通过五阶段生物发酵得NK细胞培养。
2.如权利要求1所述的混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，其特征在于，步骤（2）中生物反应器的容积为5L。
3.如权利要求1所述的混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，其特征在于，步骤（2）中连续培养从5L到1000L生物反应器，称重与分配，喂养培养基与接种物。
4.如权利要求1所述的混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，其特征在于，八种菌株为嗜酸乳杆菌、干酪乳杆菌、植物乳杆菌、保加利亚乳酸杆菌、发酵乳杆菌、乳酸双歧杆菌、两歧双歧杆菌和酵母菌。
5.如权利要求1所述的混合菌株发酵制备富含有益生菌和NK细胞的基底液的方法，其特征在于，发酵媒介还可选取番木瓜果实及叶子、生糖、RO纯净水的混合物。
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