CN112410229B - 一种富含总黄酮的羊肚菌菌丝体的制备方法 - Google Patents
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Abstract
本发明提供了一种富含总黄酮的羊肚菌固态发酵方法。本发明取新鲜白果破碎后经UV‑C辐照后孵育，添加无机盐，杀菌后接种羊肚菌种子，固态发酵后制得；其调整培养基组成后接种羊肚菌，经发酵后分离菌丝体，得到富含总黄酮的羊肚菌菌丝体；该发明利用白果富集总黄酮，再经羊肚菌发酵转化，获得富含总黄酮的新型羊肚菌菌丝，该羊肚菌发酵周期较短，富含总黄酮的羊肚菌菌丝体。
Description
技术领域
本发明属于食品生物技术领域，涉及一种富含总黄酮的羊肚菌菌丝体的制备方法。
背景技术
羊肚菌（Morchella spp.）富含蛋白质及氨基酸，其中必需氨基酸占有较大的比例，脂肪含量较低，但多为对人体有利的不饱和脂肪酸；含有较高的多糖、纤维素、矿物质。羊肚菌中含有一定量的甾醇和黄酮类等功能性成分；还含有抑制肿瘤的活性成分，具有增强免疫力、抗病毒和抗疲劳等功效。羊肚菌在食品、药品和保健品等有着广阔的开发前景。
白果是银杏种仁的俗称，白果养生延年，在宋代被列为皇家贡品。据《本草纲目》记载：“熟食温肺、益气、定喘嗽、缩小便、止白浊；生食降痰、消毒杀虫”。现代科学证明：银杏种仁含有银杏黄酮、银杏酚酸等有效成分，具有降低血清胆固醇，扩张冠状动脉和抑菌杀菌作用，是一种药食两用资源。
现有的羊肚菌发酵一般以子实体为培养目标，发酵周期较长，限制了羊肚菌的开发利用。本研究发明中提出以菌丝体为目标，开发一种富含总黄酮的羊肚菌菌丝体，发酵周期短，该羊肚菌在食品、药品和保健品等领域有着广阔的应用前景。
发明内容
本发明的目的是针对现有技术的不足之处，提供了一种富含总黄酮的羊肚菌菌丝体的制备方法。
本发明以新鲜白果为原料，经破碎后打浆经紫外线UV-C辐照，孵育后，再添加无机盐，杀菌后接种羊肚菌种子培养基，固态发酵后分离菌丝体，得到富含总黄酮的羊肚菌菌丝体。
所述新鲜白果指采收后在-20℃保存并未做干燥处理的白果。
所述UV-C辐照10-30min，20-30℃孵育6-12h。
所述添加无机盐为氯化铵0.01%-0.1%，121℃灭菌20min。
所述羊肚菌种子培养基是在液体培养基中接种羊肚菌菌块，接种量6%-10%。培养4-6d得到。
所述液体培养基为：UV-C辐照后孵育的白果粉200-300g，葡萄糖10-20g，氯化铵0.5 -1.0g，蒸馏水1000-2000mL，121℃灭菌10-20min。
所述固态发酵中的羊肚菌固态发酵件为：接种量10%，发酵温度27℃，发酵时间12-18d；发酵结束后，经4层尼龙布过滤并洗涤，滤出物即为羊肚菌菌丝体，经烘干得到富含总黄酮的羊肚菌菌丝体。
有益效果
（1）新鲜白果经UV-C辐照孵育后，黄酮类化合物含量得到了富集；
（2）以UV-C辐照孵育后的白果为固态培养基，营养丰富，富含黄酮类化合物，稍作调整就可用于羊肚菌发酵；
（3）本发明中以氯化铵补充无机盐，有利于羊肚菌菌丝转化、富集总黄酮。
具体实施方案
下面结合实施例，对本发明进一步的说明。
实施例1
（1）按UV-C辐照后孵育的白果粉200g，葡萄糖10g，氯化铵0.5 g，蒸馏水1000mL配置液体培养基，在121℃灭菌15min。在该液态种子培养基中接种羊肚菌菌块，接种量为：羊肚菌菌块为液态种子培养基重量的6%。置于旋转摇床160rpm，培养温度为28℃，培养4d得到羊肚菌种子培养基。
（2）500g新鲜白果破碎后在后经UV-C辐照10min，28℃孵育6h，添加0.01%氯化铵，121℃灭菌20min，接种10%羊肚菌种子培养基，发酵温度27℃，发酵时间18d，发酵结束后，液体经4层尼龙布过滤并洗涤，滤出物即为羊肚菌菌丝体，经烘干得到富含总黄酮的羊肚菌菌丝体。
（3）上述羊肚菌菌丝体生物量达到0.32g/g，总黄酮含量达到40.08mg/g。
实施例2
（1）按UV-C辐照后孵育的白果粉150g，葡萄糖15g，氯化铵0.5 g，蒸馏水1000mL配置液体培养基，在100℃灭菌10min。在该液态种子培养基中接种羊肚菌菌块，接种量8%，置于旋转摇床140rpm，培养温度为26℃，培养5d得到羊肚菌种子培养基。
（2）500g新鲜白果破碎后在后经UV-C辐照10min，25℃孵育6h，添加0.05%氯化铵，121℃灭菌20min，接种10%羊肚菌种子培养基，发酵温度27℃，发酵时间15d，发酵结束后，液体经4层尼龙布过滤并洗涤，滤出物即为羊肚菌菌丝体，经烘干得到富含总黄酮的羊肚菌菌丝体。
（3）上述羊肚菌菌丝体生物量达到0.36g/g，总黄酮含量达到41.25mg/g。
实施例3
（1）按UV-C辐照后孵育的白果粉300g，葡萄糖20g，氯化铵1.0g，蒸馏水2000mL配置液体培养基，在150℃灭菌20min。在该液态种子培养基中接种羊肚菌菌块，接种量10%。置于旋转摇床200rpm，培养温度为30℃，培养6d得到羊肚菌种子培养基。
（2）500g新鲜白果破碎后在后经UV-C辐照10min，30℃孵育12h，添加0.1%氯化铵，121℃灭菌20min，接种10%羊肚菌种子培养基，发酵温度27℃，发酵时间12d，发酵结束后，液体经4层尼龙布过滤并洗涤，滤出物即为羊肚菌菌丝体，经烘干得到富含总黄酮的羊肚菌菌丝体。
上述羊肚菌菌丝体生物量达到0.38g/g，总黄酮含量达到43.72mg/g。
鉴于羊肚菌和白果的食用和药用价值，将羊肚菌和白果相结合，采用固态发酵工艺生产羊肚菌菌丝，富集羊肚菌中总黄酮，同时缩短发酵周期，开发一种具有营养和保健于一体的羊肚菌菌丝体，提升了羊肚菌的加工利用价值，为白果的开发利用开辟新的途径。
本发明利用白果富集总黄酮，再经羊肚菌发酵转化，获得富含总黄酮的新型羊肚菌菌丝；该羊肚菌发酵周期较短，富含总黄酮的羊肚菌菌丝体，在食品、药品和保健品等领域有着广阔的应用前景。
Claims (4)
1.一种富含总黄酮的羊肚菌菌丝体的制备方法，其特征在于所述方法包括以下步骤：
取新鲜白果破碎后经UV-C辐照后孵育，添加无机盐，杀菌后接种羊肚菌种子培养基，固态发酵后制得；
所述羊肚菌种子培养基是在液体培养基中接种羊肚菌菌块, 接种量6%-10%，培养4-6d得到；
所述液体培养基为：UV-C辐照后孵育的白果粉200-300g，葡萄糖10-20g，氯化铵0.5 -1.0g，蒸馏水1000-2000mL，121℃灭菌10-20min；
所述固态发酵条件为：接种量10%，发酵温度27℃，发酵时间12-18d；发酵结束后，经4层尼龙布过滤并洗涤，滤出物即为羊肚菌菌丝体，经烘干得到富含总黄酮的羊肚菌菌丝体。
2.根据权利要求1所述的富含总黄酮的羊肚菌菌丝体的制备方法，其特征在于：所述新鲜白果指采收后在-20℃保存并未做干燥处理的白果。
3.根据权利要求1所述的富含总黄酮的羊肚菌菌丝体的制备方法，其特征在于：所述UV-C辐照10-30min，20-30℃孵育6-12h。
4.根据权利要求1所述的富含总黄酮的羊肚菌菌丝体的制备方法，其特征在于：所述添加无机盐为氯化铵0.01%-0.1%，121℃灭菌20min。
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| CN103952451B (zh) * | 2014-04-29 | 2016-05-11 | 甘肃省科学院生物研究所 | 采用黄芪提取液提高羊肚菌深层发酵菌丝体多糖含量的方法 |
| CN105838620A (zh) * | 2016-03-28 | 2016-08-10 | 北京七巧时代科技有限公司 | 一种灵芝培养基以及灵芝的培养方法 |
| CN105925638A (zh) * | 2016-06-06 | 2016-09-07 | 江苏大学 | 一种利用银耳同时制备银杏内酯b和槲皮素的方法 |
| CN107267403A (zh) * | 2017-08-02 | 2017-10-20 | 房玉玺 | 羊肚菌菌丝体的培养基、羊肚菌复合菌丝体及制备方法 |
| CN109504613A (zh) * | 2019-01-23 | 2019-03-22 | 上海市农业科学院 | 一种羊肚菌菌丝体的培养方法及其培养基 |
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