CN115927096A - 一种具有降尿酸功效的药食同源酵素及其制备方法 - Google Patents
一种具有降尿酸功效的药食同源酵素及其制备方法 Download PDFInfo
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Abstract
本发明属于酵素技术领域，具体涉及一种具有降尿酸功效的药食同源酵素及其制备方法，一种具有降尿酸功效的药食同源酵素的制备方法包括以下步骤：菌种培养；接种发酵；陈酿：最后将发酵完的液体倒入储存罐里进行常温密闭陈酿180d，制得药食同源酵素。本发明利用微生物发酵技术，把益生菌和药食同源物质相结合，将药食同源物质中的有效成分进行发酵转换，真正做到了食药，同时本发明黄嘌呤氧化酶抑制率可达91.39％，它的降尿酸效果达到了0.3g/L别嘌呤醇药物的疗效，因此，不仅从真正意义上保证了降尿酸的效果，并且无毒副作用，可长期食用。
Description
技术领域
本发明涉及酵素技术领域，具体为一种具有降尿酸功效的药食同源酵素及其制备方法。
背景技术
酵素又称“酶”，随着生活水平的提高，人们的保健意识逐渐增强，酵素越来越受到人们的青睐，酵素存在于所有的活细胞内，是维持机体正常功能、修复组织等生命活动的必不可少的物质。酵素按用途可分为食用酵素、环保酵素、日化酵素、农用酵素及饲用酵素。其中，食用酵素是指以一种或多种新鲜蔬菜、水果、菌菇、药食同源中草药等为原料，经多种有益菌发酵而成的功能性产品，含有丰富的酶、维生素、矿物质、多糖类、寡糖类、蛋白质及多肽类、氨基酸类、维生素类及醇、酯、酸、酚类等营养成分，具有改善肠胃功能、增强机体免疫力、美白抗衰等功能。
随着我国经济的发展，人民生活水平、营养条件和工作压力不断提高，“第四高”——高尿酸正悄然威胁人们健康。近年来，高尿酸血症患病率逐年升高，正在逼近传统“三高”。在日本，1995年厚生省的调查结果显示，高尿酸血症在成男性中的患病率已占25.0％；在欧美地区高尿酸血症患病率为2.0～18.0％。20世纪80年代初，有学者在国内进行的人群调查显示，高尿酸血症患病率男性为1.4％，女性为1.3％；进入21世纪，高尿酸血症患病率已达到7.6％～21.8％，并随年龄增长而增长，中老年人患病率可达25.6％，即我国高尿酸血症患者已逼近1亿，数值紧追糖尿病0.92亿、高血压2亿、血脂异常2亿的发病率。高尿酸血症可引发多种危险疾病。高尿酸血症可引起关节痛风、会危害肾脏、促使动脉硬化、引起高血压，并常伴随高血糖、高血脂。
目前痛风治疗的药物主要包括三类，即传统中药、西药以及多种中药混合在一起形成的中成药。常见的西药包括别嘌呤醇、苯溴马隆及非布司他等，它们能够有效抑制XO酶活性，进而抑制尿酸的合成，但这些药物的使用，往往存在肠胃不适、皮肤发热、红肿及瘙痒等副作用，偶尔的使用可能并不会出现不适的反应，但长期的使用就会陷入“温水煮青蛙”的危险之中。因此治疗痛风的药食同源植物受到国内外研究者的重视。
药食同源，也称食药物质，是指传统作为食品，且列入《中华人民共和国药典》的物质。
发酵药食同源技术是将天然药食同源食材，通过微生物发酵的方法，对提取的有效成分进行生物学转化，将其中的大分子物质，经过微生物转化成为能够被人体肠道直接吸收的小分子成分，使其中的药效成分快速吸收、定量疗效的新型技术。
发明内容
针对相关技术中的问题，本发明提出的一种具有降尿酸功效的药食同源酵素及其制备方法，以克服现有相关技术所存在的上述技术问题，本发明利用微生物发酵技术，把益生菌和药食同源物质相结合，将药食同源物质中的有效成分进行发酵转换，真正做到了食药，同时本发明黄嘌呤氧化酶抑制率可达91.39％，它的降尿酸效果达到了0.3g/L别嘌呤醇药物的疗效，因此，不仅从真正意义上保证了降尿酸的效果，并且无毒副作用，可长期食用。
为实现上述目的，本发明提供如下技术方案：
本发明提供一种具有降尿酸功效的药食同源酵素的制备方法，包括以下步骤：
(1)菌种培养
①、菌种：嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119、植物乳杆菌植物亚种CICC20267、巴氏醋酸菌CICC 20002、异常威克汉姆酵母CICC 1312；
②、种子培养基：MRS肉汤培养基、麦芽汁培养基、葡萄糖琼脂培养基；
所述巴氏醋酸菌CICC 20002采用葡萄糖琼脂培养基在30℃的条件下有氧发酵培养30h,所述异常威克汉姆酵母CICC 1312采用麦芽汁培养基在28℃的条件下有氧发酵培养36h,所述嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119和植物乳杆菌植物亚种CICC20267均采用MRS肉汤培养基在37℃的条件下无氧发酵24h；
(2)接种发酵
将培养好的菌种(嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119、植物乳杆菌植物亚种CICC20267、巴氏醋酸菌CICC20002、异常威克汉姆酵母CICC 1312)接种到发酵培养基中，发酵条件为：有氧发酵3d、35℃、搅拌，接着转厌氧发酵3d、30℃；
(3)陈酿
最后将发酵完的液体倒入储存罐里进行常温密闭陈酿180d，制得药食同源酵素。
优选的，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花5-10g/L、嘉宝果5-10g/L、葵花盘肽5-10g/L、山药1-3g/L、沙棘3-5g/L、五指毛桃1-4g/L、百合0.5-1.5g/L、罗汉果1-2g/L、刺梨1-3g/L、蔗糖15-25g/L。
优选的，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花6-8g/L、嘉宝果6-8g/L、葵花盘肽6-8g/L、山药2-3g/L、沙棘3-4g/L、五指毛桃2-3g/L、百合0.6-0.9g/L、罗汉果1-2g/L、刺梨1-3g/L、蔗糖16-22g/L。
优选的，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花7g/L、嘉宝果6g/L、葵花盘肽8g/L、山药2g/L、沙棘3g/L、五指毛桃2g/L、百合0.7g/L、罗汉果2g/L、刺梨2g/L、蔗糖20g/L。
菊花是一种重要的药食同源植物,具有强黄嘌呤氧化酶抑制(XOI)活性以及改善、缓解高尿酸血症的潜力,是开发具有降尿酸活性植物性健康食品的重要来源。
嘉宝果：嘉宝果又名珍宝果、树葡萄，是桃金娘科拟爱神木属的植物。嘉宝果富含碳水化合物、膳食纤维、有机酸、维生素、矿质元素、色素等成分，研究表明，嘉宝果提取物具有降尿酸、降血脂等作用。并且嘉宝果气味芬芳，是酵素领域的优良原料。
葵花盘肽：葵花盘为除葵花子后向日葵盘，《河北中草药》曰：“葵花盘清热燥湿，舒气散结，治头痛、头晕、风火牙痛，目赤云翳，颈前腹痛，外用于蜂窝组织炎。”具有明显消炎、利尿、降压的作用。葵花盘肽是葵花盘中具有降尿酸和溶解痛风石的成分，但是传统中药加工方法不能使葵花盘有效成分充分发挥药效，我们采用的是酶水解法制备的葵花盘小分子肽。
本发明提供还一种具有降尿酸功效的药食同源酵素，通过以上的任一项所述的制备方法获得。
与现有技术相比，本发明的有益效果是：
(1)本发明为一种具有降尿酸功效的药食同源酵素及其制备方法，本发明利用微生物发酵技术，把益生菌和药食同源物质相结合，将药食同源物质中的有效成分进行发酵转换，真正做到了食药；
(2)本发明为一种具有降尿酸功效的药食同源酵素及其制备方法，本发明的药食同源酵素，其黄嘌呤氧化酶抑制率可达91.39％，它的降尿酸效果达到了0.3g/L别嘌呤醇药物的疗效，因此，不仅从真正意义上保证了降尿酸的效果，并且无毒副作用，可长期食用。
附图说明
图1是本发明的空白组的高效液相色谱图；
图2-图7是本发明不同浓度的尿酸标准溶液的高效液相色谱图；
图8是图2-7所得出的尿酸标准曲线；
图9和图10是药食同源酵素的高效液相色谱图；
图11和图12是0.1g/L别嘌呤醇标准液的高效液相色谱图；
图13和图14是0.3g/L别嘌呤醇标准液的高效液相色谱图；
图15是图5的质控高效液相色谱图。
具体实施方式
下面将结合本发明实施例，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其它实施例，都属于本发明保护的范围。
实施例
本发明提供一种具有降尿酸功效的药食同源酵素的制备方法，包括以下步骤：
(1)菌种培养
①、菌种：嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119、植物乳杆菌植物亚种CICC20267、巴氏醋酸菌CICC 20002、异常威克汉姆酵母CICC 1312；
②、种子培养基：MRS肉汤培养基、麦芽汁培养基、葡萄糖琼脂培养基；
所述巴氏醋酸菌CICC 20002采用葡萄糖琼脂培养基在30℃的条件下有氧发酵培养30h,所述异常威克汉姆酵母CICC 1312采用麦芽汁培养基在28℃的条件下有氧发酵培养36h,所述嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119和植物乳杆菌植物亚种CICC20267均采用MRS肉汤培养基在37℃的条件下无氧发酵24h；
(2)接种发酵
将培养好的菌种(嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119、植物乳杆菌植物亚种CICC20267、巴氏醋酸菌CICC20002、异常威克汉姆酵母CICC 1312)接种到发酵培养基中，发酵条件为：有氧发酵3d、35℃、搅拌，接着转厌氧发酵3d、30℃；
(3)陈酿
最后将发酵完的液体倒入储存罐里进行常温密闭陈酿180d，制得药食同源酵素。
进一步的，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花5-10g/L、嘉宝果5-10g/L、葵花盘肽5-10g/L、山药1-3g/L、沙棘3-5g/L、五指毛桃1-4g/L、百合0.5-1.5g/L、罗汉果1-2g/L、刺梨1-3g/L、蔗糖15-25g/L。
进一步的，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花6-8g/L、嘉宝果6-8g/L、葵花盘肽6-8g/L、山药2-3g/L、沙棘3-4g/L、五指毛桃2-3g/L、百合0.6-0.9g/L、罗汉果1-2g/L、刺梨1-3g/L、蔗糖16-22g/L。
进一步的，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花7g/L、嘉宝果6g/L、葵花盘肽8g/L、山药2g/L、沙棘3g/L、五指毛桃2g/L、百合0.7g/L、罗汉果2g/L、刺梨2g/L、蔗糖20g/L。
本发明提供一种具有降尿酸功效的药食同源酵素，通过以上的任一项所述的制备方法获得。
降尿酸高效液相色谱实验
降尿酸高效液相色谱实验委托广东省中药研究所检测中心进行检测实验。本实验通过高效液相色谱仪测定药食同源酵素的黄嘌呤氧化酶的抑制率，通过抑制率的大小判断降尿酸的效果，同时对照组采用不同浓度的别嘌呤醇(痛风常用药)。
仪器参数：高效液相色谱仪(GDTCMEA01-02),色谱柱：SyncronisC18250×4.6mm 5μm(GDTCM-Z-Y-15-C18)；柱温条件：25℃；流动相：15mmol/L磷酸二氢铵-甲醇(95：5)；检测器：二极管阵列检测器(DAD)；波长：290nm。进样量：20μl；流速：1.0ml/min。
样品反应条件(总体积3600μL)：10mL离心管中依次加入200μL样品，50μL的0.03U/mL XO酶液，混匀，37℃下温浴10min，加入400μL0.42mM的黄嘌呤溶液，2800μLTris-HCl缓冲液启动反应，30℃下保温20min，加150μL1MHCl终止反应。使用去离子水代替样品做空白实验。过0.22μm有机滤膜过滤反应液后进行高相液相检测。
使用不同浓度的尿酸标准溶液建立标准曲线，Y＝1.3538X(R2＝0.9999)(其中X代表尿酸浓度单位为μg/mL，Y表示尿酸的峰面积)。XO抑制活性计算方法为：
抑制率(％)＝(C0-C)/C0*100％(C0空白浓度、C测试浓度)
进样名称S1-S6是不同浓度的尿酸标准溶液色谱图；
202209101F001-1和202209101F001-2是药食同源酵素样品；
202209101F002-1和202209101F002-2是0.1g/L别嘌呤醇样品；
202209101F003-1和202209101F003-2是0.3g/L别嘌呤醇样品。
药食同源酵素的黄嘌呤氧化酶的抑制率为91.39％，0.1g/L的别嘌呤醇的黄嘌呤氧化酶的抑制率为85.53％，0.3g/L的别嘌呤醇的黄嘌呤氧化酶的抑制率为97.89％，与别别嘌呤醇相比较，药食同源酵素不仅具有同样高的降尿酸效果，并且药食同源酵素不是药品，可以长期服用，没有副作用。
色谱图请参阅图1所示。
| 进样名称：S1 | 运行时间(min)：10.00 |
| 瓶号：GA2 | 进样量：20.00 |
| 进样类型:校正标准品 | 通道：UV-VIS-1 |
| 校正级别：1 | 波长：n.a. |
| 仪器方法：尿酸-10min | 带宽：n.a. |
| 处理方法：防腐剂处理方法 | 稀释因子：1.0000 |
| 进样时间：2022/09/3020：10 | 样品重量：1.0000 |
色谱图请参阅图2所示。
色谱图请参阅图3所示。
色谱图请参阅图4所示。
色谱图请参阅图5所示。
色谱图请参阅图6所示。
色谱图请参阅图7所示。
校正图请参阅图8所示。
色谱图请参阅图9所示。
色谱图请参阅图10所示。
色谱图请参阅图11所示。
色谱图请参阅图12所示。
色谱图请参阅图13所示。
色谱图请参阅图14所示。
色谱图请参阅图15所示。
通过上述实验可得知，本发明利用微生物发酵技术，把益生菌和药食同源物质相结合，将药食同源物质中的有效成分进行发酵转换，真正做到了食药；本发明的药食同源酵素，其黄嘌呤氧化酶抑制率可达91.39％，它的降尿酸效果达到了0.3g/L别嘌呤醇药物的疗效，因此，不仅从真正意义上保证了降尿酸的效果，并且无毒副作用，可长期食用。
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。
Claims (5)
1.一种具有降尿酸功效的药食同源酵素的制备方法，其特征在于，包括以下步骤：
(1)菌种培养
①、菌种：嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119、植物乳杆菌植物亚种CICC20267、巴氏醋酸菌CICC 20002、异常威克汉姆酵母CICC 1312；
②、种子培养基：MRS肉汤培养基、麦芽汁培养基、葡萄糖琼脂培养基；
所述巴氏醋酸菌CICC 20002采用葡萄糖琼脂培养基在30℃的条件下有氧发酵培养30h,所述异常威克汉姆酵母CICC 1312采用麦芽汁培养基在28℃的条件下有氧发酵培养36h,所述嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119和植物乳杆菌植物亚种CICC20267均采用MRS肉汤培养基在37℃的条件下无氧发酵24h；
(2)接种发酵
将培养好的菌种(嗜酸乳杆菌CICC6096、青春双歧杆菌CICC6180、鼠李糖乳杆菌CICC23119、植物乳杆菌植物亚种CICC20267、巴氏醋酸菌CICC20002、异常威克汉姆酵母CICC 1312)接种到发酵培养基中，发酵条件为：有氧发酵3d、35℃、搅拌，接着转厌氧发酵3d、30℃；
(3)陈酿
最后将发酵完的液体倒入储存罐里进行常温密闭陈酿180d，制得药食同源酵素。
2.根据权利要求1所述的一种具有降尿酸功效的药食同源酵素的制备方法，其特征在于，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花5-10g/L、嘉宝果5-10g/L、葵花盘肽5-10g/L、山药1-3g/L、沙棘3-5g/L、五指毛桃1-4g/L、百合0.5-1.5g/L、罗汉果1-2g/L、刺梨1-3g/L、蔗糖15-25g/L。
3.根据权利要求2所述的一种具有降尿酸功效的药食同源酵素的制备方法，其特征在于，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花6-8g/L、嘉宝果6-8g/L、葵花盘肽6-8g/L、山药2-3g/L、沙棘3-4g/L、五指毛桃2-3g/L、百合0.6-0.9g/L、罗汉果1-2g/L、刺梨1-3g/L、蔗糖16-22g/L。
4.根据权利要求3所述的一种具有降尿酸功效的药食同源酵素的制备方法，其特征在于，所述步骤(2)中的发酵培养基的成分及其含量包括:菊花7g/L、嘉宝果6g/L、葵花盘肽8g/L、山药2g/L、沙棘3g/L、五指毛桃2g/L、百合0.7g/L、罗汉果2g/L、刺梨2g/L、蔗糖20g/L。
5.一种具有降尿酸功效的药食同源酵素，通过权利要求1-4任一项所述的制备方法获得。
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| CN116570640A (zh) * | 2023-07-12 | 2023-08-11 | 清枫链食苏打饮品(吉林)有限公司 | 葵花盘生物碱及衍生物在降尿酸溶解痛风石产品中的应用 |
| CN117099890A (zh) * | 2023-09-28 | 2023-11-24 | 泰昊乐生物科技有限公司 | 一种促进降尿酸、降血糖血脂的无色无味低聚肽液体饮料及其加工方法 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116570640A (zh) * | 2023-07-12 | 2023-08-11 | 清枫链食苏打饮品(吉林)有限公司 | 葵花盘生物碱及衍生物在降尿酸溶解痛风石产品中的应用 |
| CN116570640B (zh) * | 2023-07-12 | 2023-09-05 | 清枫链食苏打饮品(吉林)有限公司 | 葵花盘生物碱及衍生物在降尿酸溶解痛风石产品中的应用 |
| CN117099890A (zh) * | 2023-09-28 | 2023-11-24 | 泰昊乐生物科技有限公司 | 一种促进降尿酸、降血糖血脂的无色无味低聚肽液体饮料及其加工方法 |
| CN117099890B (zh) * | 2023-09-28 | 2024-03-26 | 泰昊乐生物科技有限公司 | 一种无色无味低聚肽液体饮料及其加工方法 |
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