CN110403858A - 细菌纤维素/普洱茶提取物复合功能材料及其制备方法 - Google Patents
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Abstract
本发明公开了一种细菌纤维素/普洱茶提取物复合功能材料及其制备方法。所述方法选取活化的细菌纤维素产生菌接入种子培养液中,动态培养后将种子培养液均匀接入含有普洱茶提取物的发酵培养基中,发酵培养,发酵产物经纯化处理、切割,得到均匀分布有普洱茶提取物的三维网络状细菌纤维素/普洱茶提取物复合材料。本发明采用独特的原位复合制备方法,制得的复合材料稳定性好且复合均匀不易流失,能够充分将细菌纤维素膜的良好的生物相容性、高保水性以及良好的敷贴性与普洱茶提取物的抗菌,抗氧化和抗辐射功能结合起来,可用于制备具有清除人体自由基、抗氧化、抗辐射等多位一体功能的营养面膜或促愈敷料等应用领域。
Description
技术领域
本发明属于生物医用技术领域,涉及一种细菌纤维素/普洱茶提取物复合功能材料及其制备方法。
背景技术
细菌纤维素(BC)是由D-吡喃葡萄糖以β-1,4-糖苷键连接而成的直链多糖。与动、植物纤维相比,细菌纤维素具有更多的优异性能,如纤维直径为10-100nm,是最细的合成纤维直径的1/10,是棉纤维和木纤维的1/1000~1/100;层层折叠的三维网状结构,持有巨大的比表面积;丝束间存在大量的羟基,与水分子间形成氢键,亲水性极强,能吸收比自身干重大60~700倍的水分,保湿效果显著。细菌纤维素具有良好的生物相容性和高效保湿性,既可作为敷料,又可作为面膜,应用于医学和护肤品领域。
对于受损皮肤,细菌纤维素作为良好的医药载体,能向患处输送治疗药物的同时隔绝患处与外界的接触,防止伤口感染,同时保证患处的湿润,易于患者康复。细菌纤维素也可以作为人工皮肤,诱导受损上皮细胞修复。Lin YK等(Lin YK,et al.Effects ofdifferent extracellular matrices and growth factor immobilization onbiodegradability and biocompatibility of macroporous bacterial cellulose[J].Journal of Bioactive and Compatible Polymers,2011,26(5):508-518.)通过物理破坏,制备微孔细菌纤维素凝胶,并修饰细胞外基质(胶原蛋白,弹性蛋白和透明质酸)和生长因子等来提高凝胶的生物适应性及生物可降解性,制得了理想的皮肤替代物。另外,细菌纤维素作为面膜使用时,由于其能自由成形并任意裁剪,且纤维直径极小,为纳米级,形成的膜敷于皮肤表面时,能与皮沟(宽度50nm)贴合更紧,在保持皮肤湿润的同时更能促进有效物质的输送。
普洱茶作为我国特有的传统名茶,内含丰富的茶多酚、茶氨酸、茶色素、茶多糖等物质,具有抗衰老、抗辐射、消除人体自由基、抑菌消炎、抗氧化、抗癌、美容益寿等功效,广泛应用于保健食品、药辅制品、日化、化工等领域。
发明内容
本发明的目的在于提供一种细菌纤维素/普洱茶提取物复合功能材料及其制备方法。
本发明的目的通过下述技术方案实现:
细菌纤维素/普洱茶提取物复合功能材料的制备方法,具体步骤如下:
步骤1,将细菌纤维素产生菌接入种子培养液中,进行动态培养;
步骤2,制备普洱茶提取物溶液:将普洱茶粉碎,加入5~10倍体积的温度为80~90℃的水,密封保温浸泡40~60分钟后过滤,所得滤渣用3~8倍体积的温度为70~90℃的水浸泡30~50分钟后过滤,所得滤渣再用1~6倍体积的温度为70~90℃的水浸泡30~50分钟后过滤,合并三次过滤的滤液,加水至配制发酵培养基所需的溶液量;
步骤3,制备发酵培养基:称取好发酵培养基后加入普洱茶提取物溶液至所需溶液量,混合均匀后真空灭菌;
步骤4,将种子培养液均匀接入含有普洱茶提取物的发酵培养基中,摇匀后倒入模具中,静态培养后取出发酵产物,发酵产物经纯化处理、切割,得到细菌纤维素/普洱茶提取物复合材料。
步骤1和步骤4中,所述的培养温度为26~32℃,种子液接入量为1%~20%。
步骤1中,所述的细菌纤维素产生菌选自木醋杆菌、根瘤菌属、八叠球菌属、假单胞菌属、无色杆菌属、产碱菌属、气杆菌属和固氮菌属中的一种或几种。
步骤2中,所述的普洱茶与发酵培养基的水的质量比为1:9~24。
步骤4中,静态培养时间可以是3~7天,根据所需复合材料厚度不同从而决定反应时间。
步骤4中,纯化方法为将发酵产物在质量百分含量为3‰的NaOH水溶液中,70~90℃的温度下加热1~4h,反复水洗至中性。
本发明还提供上述制备方法制得的细菌纤维素/普洱茶提取物复合材料。
与现有技术相比,本发明具有以下优点:
本发明采用原位复合的方法,普洱茶提取物均匀分布在细菌纤维素的三维网络状结构的孔隙中,使得两者结合牢固不易流失,具有良好的稳定性。所用原料及产物均具有安全环保的特点,制备过程简单易行、操作方便,并且有效地将细菌纤维素的优良的保水保湿性能,良好的生物相容性等优点和普洱茶提取物的抗衰老、抗辐射、消除人体自由基、抑菌消炎、抗氧化等特点充分结合起来,可用于制备具有清除人体自由基、抗氧化、抗辐射等多位一体功能的营养面膜或促愈敷料等。
附图说明
图1为本发明的细菌纤维素/普洱茶提取物复合材料的实物图。
图2为本发明的细菌纤维素的扫描电镜图。
图3为本发明的细菌纤维素/普洱茶提取物复合材料的扫描电镜图。
图4为本发明的细菌纤维素/普洱茶提取物复合材料的红外谱图。
具体实施方式
下面结合具体实施例和附图对本发明做进一步说明。
实施例1
步骤1,活化的木醋杆菌菌株接入50ml种子培养液中,26℃温度下进行动态培养;
步骤2,制备普洱茶提取物溶液:将10g普洱茶粉碎,加入50ml的温度为70℃的蒸馏水,密封保温浸泡40分钟后过滤,所得滤渣用30ml的温度为70℃的蒸馏水浸泡30分钟后过滤,所得滤渣再用10ml的温度为70℃的蒸馏水浸泡30分钟后过滤,合并三次过滤的滤液,加蒸馏水至250ml。
步骤3,制备发酵培养基:称取好发酵培养基后加入250ml普洱茶提取物溶液,混合均匀后真空灭菌;
步骤4,将2.5ml种子培养液均匀接入含有普洱茶提取物的发酵培养基中,摇匀后倒入模具中26℃温度下静态培养7天后取出发酵产物。发酵产物在质量百分含量为3‰的NaOH水溶液中,在70℃的温度下加热4小时。再用蒸馏水反复冲洗至中性、切割,得到细菌纤维素/普洱茶提取物复合功能材料。
实施例2
步骤1,选取活化的木醋杆菌菌株接入50ml种子培养液中,32℃温度下进行动态培养。
步骤2,制备普洱茶提取物溶液:将10g普洱茶粉碎,加入100ml的温度为90℃的蒸馏水,密封保温浸泡60分钟后过滤,所得滤渣用80ml的温度为90℃的蒸馏水浸泡50分钟后过滤,所得滤渣再用60ml的温度为90℃的蒸馏水浸泡50分钟后过滤,合并三次过滤的滤液,加蒸馏水至250ml。
步骤3,制备发酵培养基:称取好发酵培养基后加入250ml普洱茶提取物溶液,混合均匀后真空灭菌;
步骤4,将50ml种子培养液均匀接入含有普洱茶提取物的发酵培养基中,摇匀后倒入模具中32℃温度下静态培养3天后取出发酵产物。发酵产物在质量百分含量为3‰的NaOH水溶液中,在90℃的温度下加热1小时。再用蒸馏水反复冲洗至中性、切割,得到细菌纤维素/普洱茶提取物复合功能材料。
实施例3
步骤1,选取活化的木醋杆菌菌株接入50ml种子培养液中,进行动态培养。
步骤2,制备普洱茶提取物溶液:将10g普洱茶粉碎,加入70ml的温度为80℃的蒸馏水,密封保温浸泡50分钟后过滤,所得滤渣用50ml的温度为80℃的蒸馏水浸泡40分钟后过滤,所得滤渣再用40ml的温度为80℃的蒸馏水浸泡40分钟后过滤,合并三次过滤的滤液,加蒸馏水至250ml。
步骤3,制备发酵培养基:称取好发酵培养基后加入250ml普洱茶提取物溶液,混合均匀后真空灭菌;
步骤4,将25ml种子培养液均匀接入含有普洱茶提取物的发酵培养基中,摇匀后倒入模具中30℃温度下静态培养5天后取出发酵产物。发酵产物在质量百分含量为3‰的NaOH水溶液中,在80℃的温度下加热2小时。再用蒸馏水反复冲洗至中性、切割,得到细菌纤维素/普洱茶提取物复合功能材料。
图1为用本方法制备得到的细菌纤维素/普洱茶提取物复合材料实物图,由图可看出该复合材料是无色透明的高保水性柔软的膜状材料,能够与皮肤很好的贴合;图3为用本方法制备得到的细菌纤维素/普洱茶提取物复合材料的扫描电镜图,由图可以看出细菌纤维素三维网状结构中大量负载了普洱茶提取物;图4为用本方法制备得到的细菌纤维素/普洱茶提取物复合材料的红外谱图,在1400cm-1和1600cm-1处出现了普洱茶提取物中茶多酚的特征谱线,且800~500cm-1指纹区该复合材料也与普洱茶提取物相契合,说明细菌纤维素中有普洱茶提取物负载上去。
对比例1
本对比例与实施例3基本相同,唯一不同的是配制发酵培养基时不添加普洱茶提取液而改为蒸馏水,得到细菌纤维素。
图2为对比例1制得的细菌纤维素的扫描电镜图,不含有普洱茶的营养物质,此图与图3形成对比,说明用本发明可以成功将普洱茶提取物与细菌纤维素复合起来。
对比例2
本对比例与实施例3基本相同,不同的是配制发酵培养基时不添加普洱茶提取液而改为蒸馏水,得到的细菌纤维素浸泡于普洱茶提取液中,后经与实施例3步骤4相同方法处理后得到细菌纤维素/普洱茶提取物复合材料,经检测发现与实施例3对比该方法制得的复合材料中普洱茶提取物大量流失。
Claims (7)
1.细菌纤维素/普洱茶提取物复合功能材料的制备方法,其特征在于,具体步骤如下:
步骤1,将细菌纤维素产生菌接入种子培养液中,进行动态培养;
步骤2,制备普洱茶提取物溶液:将普洱茶粉碎,加入5~10倍体积的温度为80~90℃的水,密封保温浸泡40~60分钟后过滤,所得滤渣用3~8倍体积的温度为70~90℃的水浸泡30~50分钟后过滤,所得滤渣再用1~6倍体积的温度为70~90℃的水浸泡30~50分钟后过滤,合并三次过滤的滤液,加水至配制发酵培养基所需的溶液量;
步骤3,制备发酵培养基:称取好发酵培养基后加入普洱茶提取物溶液至所需溶液量,混合均匀后真空灭菌;
步骤4,将种子培养液均匀接入含有普洱茶提取物的发酵培养基中,摇匀后倒入模具中,静态培养后取出发酵产物,发酵产物经纯化处理、切割,得到细菌纤维素/普洱茶提取物复合材料。
2.根据权利要求1所述的制备方法,其特征在于,步骤1和步骤4中,所述的培养温度为26~32℃,种子液接入量为1%~20%。
3.根据权利要求1所述的制备方法,其特征在于,步骤1中,所述的细菌纤维素产生菌选自木醋杆菌、根瘤菌属、八叠球菌属、假单胞菌属、无色杆菌属、产碱菌属、气杆菌属和固氮菌属中的一种或几种。
4.根据权利要求1所述的制备方法,其特征在于,步骤2中,所述的普洱茶与发酵培养基的水的质量比为1:9~24。
5.根据权利要求1所述的制备方法,其特征在于,步骤4中,静态培养时间为3~7天。
6.根据权利要求1所述的制备方法,其特征在于,步骤4中,纯化方法为将发酵产物在质量百分含量为3‰的NaOH水溶液中,70~90℃的温度下加热1~4h,反复水洗至中性。
7.根据权利要求1至6任一所述的制备方法制得的细菌纤维素/普洱茶提取物复合材料。
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