CN111807939A - 一种具有变形链球菌抑制能力的羊栖菜多酚及制备方法 - Google Patents
一种具有变形链球菌抑制能力的羊栖菜多酚及制备方法 Download PDFInfo
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Abstract
本发明公开了一种具有变形链球菌抑制能力的羊栖菜多酚及其制备方法。该多酚类化合物包括5个苯环和14个酚羟基,且分子量为654u。其制备方法包括步骤:将干燥的羊栖菜粉碎后使用石油醚萃取12小时,利用滤纸过滤后收集固体;向所述固体中加入50%乙醇水溶液,利用摇床在25℃,转速60转/分钟条件下提取,提取结束后收集提取液;将提取液在35℃条件下旋转蒸馏8小时;加入至具有亲水、亲脂性的色谱柱中,用洗脱剂A洗脱除去分子量小于500Da的小分子酚类物质,再用洗脱剂B洗脱并收集组分,干燥后即得所述多酚类化合物。本发明中涉及的多酚类化合物,既能有效抑制变形链球菌的生长,又能减少生物膜的形成,因而可以对龋齿起到有效的治疗作用。
Description
技术领域
本发明涉及生物医药技术领域,特别涉及一种由羊栖菜中分离得到的多酚类化合物,以及制备该多酚类化合物的方法。
背景技术
我国人群的口腔健康状况不容乐观,尤其龋齿患病率一直呈现逐年上升的趋势。由龋齿引发的牙周炎、口臭等并发症严重影响了人们的身体健康和社会交往。早在1924年,人们就在龋齿患者口腔中发现了一种能在不同培养基中呈现出不同形态的细菌,并将其命名为变形链球菌。直到上世纪八十年代左右,才逐渐明确变形链球菌引发龋齿的详细途径(Thaís Manzano P.et al.Early Childhood Caries and Mutans Streptococci:ASystematic Review.Oral Health&Preventive Dentistry,2010,8(1):59-70)。
龋齿发生的全部过程可以概括如下:首先,变形链球菌粘附在牙釉质表面;随后,变形链球菌自身通过代谢分泌出细胞外基质,将饮食中的碳水化合物代谢为有机酸并形成生物膜;变形链球菌持续生长,生物膜持续酸化,将牙釉质脱矿并伴随牙齿有机质的降解,最终发生龋齿(MaiaRibeiro L.G.et al.The effect of different formulations ofchlorhexidine in reducing levels of mutans streptococci in the oral cavity:Asystematic review of the literature.Journal of Dentistry,2007,35(5):359-370.)。由此可见,龋齿的发生过程包含两个关键因素:1)变形链球菌的增殖和代谢;2)生物膜的蓄积和增长。
针对龋齿的防治,目前主要采取物理、生物或者化学手段,达到清除生物膜或抑制变形链球菌生长的目的。物理清除法包括刷牙、低水平直流电协同、光动力疗法(PDT)等,但物理疗法因疗程长且疗效受到患者依从性影响,因而实际实施效果受到限制。生物疗法是利用益生菌与致病菌之间的拮抗作用,达到抑制致病菌增殖的目的。已知有效的益生菌包括:乳杆菌属(包括嗜酸乳杆菌、干酪乳杆菌、副干酪乳杆菌等),双歧杆菌属(包括两歧双歧杆菌、短双歧杆菌、婴儿双歧杆菌、长双歧杆菌等)、酵母菌属(布拉酵母菌)、链球菌属(唾液链球菌嗜热亚种)、以及其他属(中间链球菌、粪肠球菌)微生物(李佳桐,等.益生菌防治口腔感染性疾病的研究进展,口腔疾病防治,2019,27(9):598-602)。但由于益生菌的增殖与其所处的环境条件密切相关,因而生物抑菌方法的实施效果非常容易受到患者饮食、吸烟、饮酒等日常生活习惯的影响。
化学清除是目前临床最常用且效果最为显著的方法,主要利用人工合成抗生素或具有抗菌功能的天然活性物质清除牙釉质表面的细菌或生物膜。口腔清洁用品和临床治疗中最常用的人工合成抗生素为氟化物和氯己定,但由于生物膜的存在,变形链球菌容易发生变异并逐渐对氟化物和氯己定产生耐药性。因此,人们逐渐将目光转向具有抑菌活性的天然产物。从黄岑、姜黄、蜂胶等中草药中发现的黄岑苷、黄岑素、姜黄素、tt-法尼醇等化合物,能够显著降低生物膜中胞外基质(包括蛋白质、多糖)的含量,除此以外,这些天然化合物还能降低变形链球菌生物膜产酸的能力,并起到破坏和消除生物膜的作用。椰子油、三羟基稀酚等化合物能够抑制细菌内ffh、gtfD、pdp基因片段的转录和表达,促使成熟时期的胞外生物膜发生脱落。部分酶类,尤其是脱氧核糖醛缩酶可以分解细胞外基质中的eDNA,抑制变形链球菌生物膜的形成,并减弱变形链球菌在牙齿表面的附着力。但这些天然物质都只能从减少生物膜的角度抑制龋齿的发生,而在直接杀灭变形链球菌方面,并没有明确的作用。
发明内容
本发明一方面公开了一种由羊栖菜中分离得到的多酚类化合物,其能够抑制、杀灭变形链球菌或减少生物膜形成。
一种多酚类化合物,是由羊栖菜中分离得到,其结构式如下所示:
该多酚类化合物以间苯三酚为结构单元,包括5个苯环和14个酚羟基,且分子量为654Da。利用核磁共振技术,明确该多酚类化合物在氘代甲醇为溶剂的条件下,该化合物的核磁共振碳谱(13CNMR)具有化学位移162ppm、159ppm、154ppm、152ppm、141ppm、140ppm、127ppm、126ppm、105ppm和98ppm;在氘代甲醇为溶剂条件下,该化合物的核磁共振氢谱(1HNMR)具有化学位移10.3ppm、9.6ppm、9.5ppm、8.7ppm和5.8ppm。
另一方面,本发明还公开了制备上述多酚类化合物的方法,具体包括步骤:
1)将干燥的羊栖菜粉碎后使用石油醚萃取12小时,利用滤纸过滤后收集固体;
2)向所述固体中加入50%乙醇水溶液,利用摇床在25℃,转速60转/分钟条件下提取,提取结束后收集提取液;
3)将提取液在35℃条件下旋转蒸馏8小时;
4)加入至具有亲水、亲脂性的色谱柱中,用洗脱剂A洗脱除去分子量小于500Da的小分子酚类物质,再用洗脱剂B洗脱并收集组分,干燥后即得所述多酚类化合物。
其中,步骤1所述粉碎是将羊栖菜粉碎成过20目筛的颗粒。羊栖菜粉干燥后的质量与石油醚的质量比为(0.2~2):1。
其中,步骤2所述羊栖菜粉干燥后的质量与50%乙醇水溶液的质量比为(0.2~2):1。所述摇床提取时间不少于12小时。
其中,步骤4所述色谱柱为Sephadex LH-20色谱柱,该色谱柱的填料体积为50~200mL,色谱柱径高比为1:(20~100)。所述洗脱剂A为浓度为10~80%的乙腈水溶液,洗脱剂A的用量为500~3000mL。所述洗脱剂B为浓度为10~80%的丙酮-水溶液,洗脱剂B的用量为100~2500mL。
与现有技术相比,本发明所述多酚类化合物中的羟基与苯环结构的含量明显高于其他已知结构的海藻多酚类化合物(袁圣亮,等.海藻多酚类化合物及其抗氧化活性研究进展.食品与发酵工业,2018.),正是这些酚羟基和苯环,赋予了羊栖菜多酚强烈的蛋白质吸附与结合能力,有效地抑制了微生物表面酶活性,从而起到了显著的变形链球菌抑制能力。
同时,该多酚类化合物的制备方法正是利用多酚“分子量越大,与蛋白质结合活性越强”的活性规律,结合分子筛与吸附-解吸附色谱原理,对分子量大的羊栖菜多酚组分进行分离纯化,,最终获得大分子量的多酚组分。
本发明中涉及的多酚类化合物,既能有效抑制变形链球菌的生长,又能减少生物膜的形成,因而可以对龋齿起到有效的治疗作用;
本发明以海产品-羊栖菜为原料,充分利用羊栖菜本身原料易得、价格低廉的特点,极为简便地制得了所述多酚类化合物,极大的降低了天然变形链球菌抑菌剂的生产成本。
附图说明
图1是实施例1所述的多酚类化合物的核磁共振碳谱;
图2是实施例1所述的多酚类化合物的核磁共振氢谱。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚,下面将结合附图对本发明作进一步地详细描述。
实施例1
将20份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为200mL,径高比为1:100),用洗脱剂A(浓度为80%的乙腈水溶液,用量为3000mL)洗脱除去简单酚,随后用洗脱剂B(浓度为40%的丙酮水溶液,用量为1000mL)洗脱,既得。产物进行核磁共振碳谱检测和核磁共振氢谱,结果分别如图1和图2所示。
实施例2
将60份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为110mL,径高比为1:50),用洗脱剂A(浓度为10%的乙腈水溶液,用量为2000mL)洗脱除去简单酚,随后用洗脱剂B(浓度为60%的丙酮水溶液,用量为2000mL)洗脱,既得。
实施例3
将200份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为170mL,径高比为1:30),用洗脱剂A(浓度为60%的乙腈水溶液,用量为1500mL)洗脱除去简单酚,随后用洗脱剂B(浓度为80%的丙酮水溶液,用量为200mL)洗脱,既得。
实施例4
将180份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为50mL,径高比为1:90),用洗脱剂A(浓度为20%的乙腈水溶液,用量为2000mL)洗脱除去简单酚,随后用洗脱剂B(浓度为50%的丙酮水溶液,用量为2000mL)洗脱,既得。
实施例5
将120份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为170mL,径高比为1:60),用洗脱剂A(浓度为40%的乙腈水溶液,用量为2500mL)洗脱除去简单酚,随后用洗脱剂B(浓度为10%的丙酮水溶液,用量为1000mL)洗脱,既得。
实施例6
将20至200份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为120mL,径高比为1:50),用洗脱剂A(浓度为50%的乙腈水溶液,用量为1000mL)洗脱除去简单酚,随后用洗脱剂B(浓度为70%的丙酮水溶液,用量为2500mL)洗脱,既得。
实施例7
将90份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为160mL,径高比为1:90),用洗脱剂A(浓度为60%的乙腈水溶液,用量为1500mL)洗脱除去简单酚,随后用洗脱剂B(浓度为50%的丙酮水溶液,用量为800mL)洗脱,既得。
实施例8
将160份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为90mL,径高比为1:60),用洗脱剂A(浓度为70%的乙腈水溶液,用量为2000mL)洗脱除去简单酚,随后用洗脱剂B(浓度为30%的丙酮水溶液,用量为200mL)洗脱,既得。
实施例9
将180份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为170mL,径高比为1:70),用洗脱剂A(浓度为80%的乙腈水溶液,用量为2500mL)洗脱除去简单酚,随后用洗脱剂B(浓度为60%的丙酮水溶液,用量为2300mL)洗脱,既得。
实施例10
将30份干燥的羊栖菜粉碎后,过20目筛,随后使用100份石油醚萃取12小时,去除脂类物质,利用滤纸过滤后收集固体,并向固体中加入50%乙醇水溶液100份,利用摇床在25℃,转速60转/分钟条件下提取12小时,提取结束后,收集液体,并将其在35℃条件下旋转蒸馏8小时除去有机溶剂,随后加入至Sephadex LH-20色谱柱中(填料体积为110mL,径高比为1:20),用洗脱剂A(浓度为10%的乙腈水溶液,用量为500mL)洗脱除去简单酚,随后用洗脱剂B(浓度为70%的丙酮水溶液,用量为100mL)洗脱,既得。
实施例11
利用高效液相色谱-质谱联用(LC-MS)对羊栖菜多酚的含量进行了测试,在BHI培养基(brainheartinfusion)中测试了多酚的最小抑菌浓度(MIC)和最小杀菌浓度(测试方法参考王少龙等.五倍子对致龋菌抑制作用的实验研究,口腔医学研究,2004,20(3):246-248.)。利用结晶紫染色法测试了添加羊栖菜多酚后生物膜产量,并以此计算了生物膜抑制率、水不溶性多糖产量抑制率,以及pH(测试方法参考江枫等,白藜芦醇对蜂房哈尼夫菌生物膜形成的影响,大连工业大学学报,2018,37(6):442-445)。结果如表1所示。
表1依据实施例制备的羊栖菜多酚样品中多酚含量,最小抑菌浓度(MIC)最小杀菌浓度(MBC)生物膜抑制率,水不溶性多糖产量抑制率和pH
可以看出,各实施例中的羊栖菜多酚对变形链球菌和生物膜有显著的抑制作用。并且最小抑菌浓度和最小杀菌浓度小于NaF,生物膜抑制率和水不溶性多糖产量抑制率明显大于NaF,变形链球菌液的pH也显著高于NaF。
Claims (10)
2.权利要求1所述多酚类化合物的制备方法,其特征在于,包括以下步骤:
1)将干燥的羊栖菜粉碎后,使用石油醚萃取12小时,利用滤纸过滤后收集固体;
2)向所述固体中加入50%乙醇水溶液,利用摇床在25℃,转速60转/分钟条件下提取,提取结束后收集提取液;
3)将提取液在35℃条件下旋转蒸馏8小时;
4)加入至具有亲水、亲脂性的色谱柱中,用洗脱剂A洗脱除去分子量小于500Da的小分子酚类物质,再用洗脱剂B洗脱并收集组分,干燥后即得所述多酚类化合物。
3.根据权利要求2所述多酚类化合物的制备方法,其特征在于,步骤1)所述萃取之前,将羊栖菜粉碎成过20目筛的颗粒。
4.根据权利要求2所述多酚类化合物的制备方法,其特征在于,所述羊栖菜粉干燥后的质量与石油醚的质量比为(0.2~2):1。
5.根据权利要求2所述多酚类化合物的制备方法,其特征在于,所述羊栖菜粉干燥后的质量与50%乙醇水溶液的质量比为(0.2~2):1。
6.根据权利要求2所述多酚类化合物的制备方法,其特征在于,步骤2所述摇床提取时间不少于12小时。
7.根据权利要求2所述多酚类化合物的制备方法,其特征在于,所述色谱柱为SephadexLH-20色谱柱。
8.根据权利要求6所述多酚类化合物的制备方法,其特征在于,所述Sephadex LH-20色谱柱的填料体积为50~200mL,色谱柱径高比为1:(20~100)。
9.根据权利要求2所述多酚类化合物的制备方法,其特征在于,所述洗脱剂A为浓度为10~80%的乙腈水溶液,洗脱剂A的用量为500~3000mL;所述洗脱剂B为浓度为10~80%的丙酮-水溶液,洗脱剂B的用量为100~2500mL。
10.权利要求1所述多酚类化合物在抑制、杀灭变形链球菌或减少生物膜形成的应用。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1075138A (en) * | 1964-05-13 | 1967-07-12 | Dow Chemical Co | A method for the preparation of polyhydroxy aromatic compositions |
US20130231396A1 (en) * | 2010-11-04 | 2013-09-05 | Abra Pharmaceutical Sárl | Novel self-assembling polyphenol-quinonoid polymer derivatives and uses thereof |
CN105193899A (zh) * | 2015-11-10 | 2015-12-30 | 桂林茗兴生物科技有限公司 | 从柿子中提取总酚的方法 |
CN110272334A (zh) * | 2019-07-03 | 2019-09-24 | 云南中烟工业有限责任公司 | 一种天然抗菌多酚类化合物及其制备方法与在电子烟中的应用 |
CN111944165A (zh) * | 2020-07-15 | 2020-11-17 | 汕头大学 | 一种多酚类酪氨酸酶抑制剂及其提取方法和应用 |
-
2020
- 2020-06-08 CN CN202010511276.1A patent/CN111807939A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1075138A (en) * | 1964-05-13 | 1967-07-12 | Dow Chemical Co | A method for the preparation of polyhydroxy aromatic compositions |
US20130231396A1 (en) * | 2010-11-04 | 2013-09-05 | Abra Pharmaceutical Sárl | Novel self-assembling polyphenol-quinonoid polymer derivatives and uses thereof |
CN105193899A (zh) * | 2015-11-10 | 2015-12-30 | 桂林茗兴生物科技有限公司 | 从柿子中提取总酚的方法 |
CN110272334A (zh) * | 2019-07-03 | 2019-09-24 | 云南中烟工业有限责任公司 | 一种天然抗菌多酚类化合物及其制备方法与在电子烟中的应用 |
CN111944165A (zh) * | 2020-07-15 | 2020-11-17 | 汕头大学 | 一种多酚类酪氨酸酶抑制剂及其提取方法和应用 |
Non-Patent Citations (4)
Title |
---|
K.-W. GLOMBITZA等: ""Antibiotics from Algae, XXXVI.1,2 Phiorotannins from the Brown Alga Cystoseira granulata"", 《PLANTA MEDICA》 * |
KARL-WERNER GLOMBITZA等: ""FUCOPHLORETHOLS FROM THE BROWN ALGA CARPQPIIYZLUM MASCHALOCARPUM"", 《PHYTOCHAMISTRY》 * |
MICHELLE S. TIERNEY等: ""UPLC-MS profiling of low molecular weight phlorotannin polymers in Ascophyllum nodosum, Pelvetia canaliculata and Fucus spiralis"", 《METABOLOMICS》 * |
周才琼等: "《食品营养学》", 30 November 2006, 中国计量出版社 * |
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