CN114668786A - 一种凝结芽胞杆菌发酵制备三七叶茶的方法 - Google Patents
一种凝结芽胞杆菌发酵制备三七叶茶的方法 Download PDFInfo
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Abstract
本发明公开了一种凝结芽胞杆菌发酵制备三七叶茶的方法，即三七叶经过萎凋、杀青、揉捻、晒青处理后，接种凝结芽胞杆菌（cillus coagulans），在40‑45℃、空气湿度65‑75%条件下发酵5‑8天后，干燥制得三七叶茶；本发明方法简单，易操作，本发明方法制得的产品相比于未接种凝结芽胞杆菌发酵制得的三七叶茶的总皂苷、总黄酮、总多酚明显增多，本发明为三七叶的利用提供了一种新的应用途径。
Description
技术领域
本发明属于三七叶茶制备技术领域，具体涉及一种利用凝结芽胞杆菌发酵制备三七叶茶的方法。
背景技术
三七系五加科人参属植物，作为传统中医常用名贵药材中国南方广泛种植，以干燥根及根茎入药，是我国名贵的中药材。具有滋补强壮、止血活血、活血化瘀和消肿止痛等功效。民间也常将其磨成三七粉冲服，以达到预防血栓的保健效果。然而，经过几百年的栽培，三七对环境的适应能力逐渐下降，且连作障碍问题日益凸显。同时，由于适宜三七生长的土壤环境较为局限，导致其药用资源不足。因此，三七叶的开发应用逐渐受到广大学者的关注。三七叶系五加科人参属植物三七的干燥茎叶，其性温，味辛，主要用于止血、消肿、定痛，治吐血、便血、外伤出血、痈肿毒疮、跌打肿痛、偏头痛等症。近年研究表明三七的茎叶可供药用，且毒副作用小。三七叶的活性成分三七叶皂苷属达玛烷型三萜皂苷，其对血液系统、心血管系统、神经系统和代谢系统作用与三七根皂苷相似。由于人们的传统习惯多以根入药，对三七茎叶资源综合利用研究很少，在三七连作障碍问题凸显的情况下，三七茎叶的高效利用具有重要的现实和长远意义。
凝结芽孢杆菌属于革兰氏阳性菌，兼性厌氧，具有极强的耐酸、耐盐、耐高温能力，最适生长温度约 45℃。与一般乳酸菌相比，凝结芽孢杆菌对营养底物需求低，培养温度高，不易染杂菌，产物中乳酸含量高，具有降低人体总胆固醇，改善高血脂症等特点。
目前未见利用凝结芽胞杆菌发酵制备三七叶茶的相关报道。
发明内容
本发明提供了一种凝结芽胞杆菌发酵制备三七叶茶的方法，该方法是将新鲜三七叶经过萎凋、杀青、揉捻、晒青处理后，接种凝结芽胞杆菌（cillus coagulans），在40-45℃、空气湿度65-75%条件下发酵5-8天后，干燥制得三七叶茶；本发明方法制得的三七叶茶清香浓郁，茶汤为金黄色，经凝结芽孢杆菌发酵后的三七叶的总皂苷含量、总酚、总黄酮含量明显增高，为三七叶的利用提供了理论基础，本发明产品在活血化瘀，预防心脑血管疾病，调整肠道菌群等方面具有潜在的应用前景。
上述方法的具体操作如下：将新鲜采摘的翠绿三七叶片放在簸箕里，然后置于阴凉通风处1.5-2.5h萎凋处理，把水汽晾干；将萎凋处理后的叶片置于250-300℃锅中翻炒至叶片含水率为40-50%；然后置于常温环境中反复人工轻揉，直至成条率在80%以上，将揉捻好的叶片平摊晾干；将凝结芽胞杆菌菌液喷洒至晾干的叶片上，混匀后在40-45℃、湿度65-75%条件下发酵5-8天后，干燥制得三七叶茶，含水率低于13%。
本发明凝结芽胞杆菌（cillus coagulans）为常规市售菌种，凝结芽胞杆菌的添加量为晒青后三七叶质量的0.01-0.02%，接种后三七叶含水率为40-60%。
本发明的优点和技术效果：
1、本发明采用凝结芽胞杆菌发酵三七叶，能够提高三七叶中的总皂苷、总黄酮、总多酚含量；具有降血脂、预防心脑血管疾病、调节肠道微生物的应用前景；
2、本发明制备工艺简单，易操作，适于工业化生产和市场推广应用，本发明为三七叶的资源化利用提供了新途径，对于三七种植业和加工企业的经济效益均具有重要的影响。
附图说明
图1为本发明方法制得的产品形态示意图。
具体实施方式
下面通过实施例对本发明作进一步详细说明，但本发明保护范围不局限于所述内容，下述实施例中使用的菌种购买自江苏微康生物科技有限公司；
实施例1：本凝结芽胞杆菌发酵制备三七叶茶的方法如下：
1、将新鲜采摘的翠绿三七叶片0.5kg放在簸箕里，然后置于阴凉通风处2h萎凋处理，把水汽晾干；将萎凋处理后的叶片置于250℃锅中翻炒至叶片含水率为50%；然后置于常温环境中反复人工轻揉，直至成条率在80%以上，将揉捻好的叶片平摊晾干；
2、凝结芽胞杆菌添加量为晾干三七叶质量的0.02%，将凝结芽胞杆菌菌粉用水溶解，将菌液均匀喷洒至晾干的叶片上，接种后三七叶含水率为40%，混匀后在45℃、湿度65-70%条件下发酵7天后，40℃干燥只含水率为10%，制得三七叶茶，三七叶茶形态见图1；
同时以没有接种凝结芽胞杆菌制备的三七叶茶作为对照，其他制备过程同上；
3、三七叶茶总酚总黄酮总皂苷的测定
将接种凝结芽胞杆菌制得的三七叶茶和未接种菌种制得的三七叶茶粉碎后，用乙醇提取2次，收集合并提取液，使用旋转蒸发仪蒸干有机溶剂，剩余液浓缩冻干，-30℃保存；
（1）总酚含量测定（TPC）
取4mg样品，用质量浓度80%的乙醇8mL溶解，溶解液离心取上清液，取1mL上清液与6mL去离子水混合，加0.5mL福林酚试剂，静置1min，加入质量浓度20%的Na2CO3溶液1.5mL，加蒸馏水定容到10mL，70℃水浴10min，冷却后于765nm处测吸光值；在相同实验条件下，以没食子酸为标准品，以标准品的浓度为横坐标，吸光度为纵坐标绘制标准曲线，根据标准曲线的回归方程计算样品总酚含量，最终结果以每克干样品中的总酚含量（mg/g）进行表示。
（2）总黄酮含量（TFC）
取8mg样品，用质量浓度80%乙醇4mL溶解，并稀释五倍，取1mL稀释液用质量浓度70%的乙醇定容至2.5mL，加入5% Na2CO3溶液0.15mL，摇匀静置5min，再加入5% NaNO2溶液0.15mL，摇匀静置5min，加入0.15mL 质量浓度10% 的Al（NO3）3溶液，摇匀静置6min，再加入1mL1mol/L NaOH溶液，混匀，用70%乙醇定容至5mL，静置30min，510nm处侧吸光值；在相同实验条件下，以芦丁为标准品，以标准品的浓度为横坐标，吸光度为纵坐标绘制标准曲线，根据标准曲线的回归方程计算样品总黄酮含量，最终结果以每克干样品中的总黄酮含量（mg/g）进行表示；
（3）总皂苷含量（TSC）
取2mg样品，用2mL 80%乙醇充分溶解，将1mL样品混合物放入管中，并使用miVac真空离心浓缩器（美国SP SCIENTIFIC）浓缩至干；再加入0.2mL质量浓度5%的香草醛-冰醋酸溶液，以及1mL高氯酸，70℃下水浴15min，然后迅速放入冰水中冷却2min，最后再加入2.8mL的冰醋酸稀释，混匀后，在510nm下测定吸光值；在相同实验条件下，以人参皂苷Re为标准品，以标准品的浓度为横坐标，吸光度为纵坐标绘制标准曲线，根据标准曲线的回归方程计算样品总皂苷含量，最终结果以每克干样品中的总皂苷含量（mg/g）进行表示；
检测结果见下表，由表可以看出相比于未接种凝结芽胞杆菌制得的三七叶茶，接种凝结芽胞杆菌制得的三七叶茶的总皂苷含量明显升高，总酚、总黄酮含量均增加；
实施例2：本凝结芽胞杆菌发酵制备三七叶茶的方法如下：
1、将新鲜采摘的翠绿三七叶片0.5kg放在簸箕里，然后置于阴凉通风处2h萎凋处理，把水汽晾干；将萎凋处理后的叶片置于300℃锅中翻炒至叶片含水率为40%；然后置于常温环境中反复人工轻揉，直至成条率在80%以上，将揉捻好的叶片平摊晾干；
2、凝结芽胞杆菌添加量为晾干三七叶质量的0.015%，将凝结芽胞杆菌菌粉用水溶解，将菌液均匀喷洒至晾干的叶片上，接种后三七叶含水率为60%，混匀后在45℃、湿度70-75%条件下发酵6天后，40℃干燥只含水率为5%，制得三七叶茶；
同时以没有接种凝结芽胞杆菌制备的三七叶茶作为对照，其他制备过程同上；
3、三七叶茶总酚总黄酮总皂苷的测定，方法同实施例1，结果见下表，由表可以看出相比于未接种凝结芽胞杆菌制得的三七叶茶，接种凝结芽胞杆菌制得的三七叶茶的总皂苷含量明显升高，总酚、总黄酮含量均增加；
上述结果显示本发明方法制得的三七叶茶具有活血化瘀、预防心脑血管疾病、调整肠道菌群等潜在的应用前景，本发明为三七叶的利用提供了一个新的途径。
Claims (5)
1.一种凝结芽胞杆菌发酵制备三七叶茶的方法，其特征在于：新鲜三七叶经过萎凋、杀青、揉捻、晒青处理后，接种凝结芽胞杆菌（cillus coagulans），在40-45℃、空气湿度65-75%条件下发酵5-8天后，干燥制得三七叶茶。
2.根据权利要求1所述的凝结芽胞杆菌发酵制备三七叶茶的方法，其特征在于：萎凋是在阴凉通风处放置1.5-2.5h。
3.根据权利要求2所述的凝结芽胞杆菌发酵制备三七叶茶的方法，其特征在于：杀青是将萎凋处理后的叶片置于250-300℃下翻炒至叶片含水率为40-50%。
4.根据权利要求3所述的凝结芽胞杆菌发酵制备三七叶茶的方法，其特征在于：揉捻是将杀青后叶片置于常温环境中反复人工轻揉，直至成条率在80%以上。
5.根据权利要求4所述的凝结芽胞杆菌发酵制备三七叶茶的方法，其特征在于：晒青是将揉捻好的茶叶平摊晾干。
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| CN108850350A (zh) * | 2018-07-09 | 2018-11-23 | 马卫军 | 一种三七叶保健茶的制备方法 |
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