CN112370458A - 一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法 - Google Patents
一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法 Download PDFInfo
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Abstract
本发明提供了一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法，其特征在于，包括如下步骤：去除魔芋粉生物碱；高浓度底物水解制备魔芋寡糖；制备魔芋寡糖胶：取魔芋寡糖粉加入40‑50％的酒精，搅拌混合，放入到微波炉中，微波强度100％，加热至溶液处于沸腾状态，悬浮液溶解后变澄清，继续加热，通过微波加热处理使魔芋寡糖浓度达到70‑80％，即形成魔芋寡糖胶，用于涂抹手臂、皮肤表面，迅速形成干燥薄膜，对外来病原形成屏障保护作用。本发明利用魔芋寡糖特性制备的魔芋寡糖胶，能修复人体细胞，抑制病原微生物的生长，涂抹细胞表面能覆膜成型，细胞包埋病原菌，抑制生长，用于人体表面皮肤的保护和抗菌作用、伤口的愈合。
Description
技术领域
本发明涉及生物制药技术领域，具体涉及一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法。
背景技术
魔芋为天南星科魔芋属，多年生草本植物的块茎，广泛分布在中国的云南、贵州、四川、湖北、安徽、河南等地，主产地以云南为主，主要种植分布于富源、玉溪、楚雄、丽江、版纳等地。新鲜魔芋原料含水量能达到83-84％。魔芋精粉主要成分为葡甘露聚糖(KonjacGlucomannan，KGM)，含量可达到70％以上，分子量为20×104～20×105(随魔芋原料来源不同而异)，因其吸水膨胀系数较大，1％的水溶液就能形成凝胶状。可用于食品稳定剂、增稠剂、胶凝剂，石油萃取黏胶剂。魔芋中的主要成份为魔芋甘露聚糖(mannan)，是非淀粉多糖中的一种，以β-1,4-D-吡喃甘露糖苷键连接的线状多糖，水解终产物为甘露糖和葡萄糖，比例为1：1.6-1.7。如果控制水解聚合程度就能形成分子大小不一的魔芋寡糖。
魔芋寡糖的制备常采用酶法水解、Y射线法、生物转化法进行，为便于工业化生产多采用酶法-酸法水解联合制备。而随着科学技术的发展，β-甘露聚糖酶成为当前用于魔芋寡糖制备的重要糖苷水解酶，是一类能水解含有甘露糖苷的甘露聚糖(包括异甘露聚糖)的内切水解酶，形成的产物为甘露单糖、二糖、甘三糖、四糖等低聚糖。而随着六碳糖单体通过糖苷键连接形成不同结构的产物，导致功能活性物质位点增多，大量的活性物质形成的生物活性部位就更加显现。魔芋寡糖的生物活性也是被众多科学家进行了研究，清理自由基、提高机体抗氧化的能力也获得了肯定。
鉴于多糖成膜过程报道不多，特别是中性多糖成膜过程的研究没有报道，而用于寡糖进行成膜的研究更基本没有报道。如何利用魔芋寡糖的功能特性，成为当前开发的新思路。
发明内容
本发明的目的是提供一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法，用于皮肤的愈合和防护作用。
本发明提供了一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法，包括如下步骤：
(1)去除魔芋粉中的生物碱
取魔芋粉，按照料水比1：2加入80％的酒精进行醇洗，搅拌混合，静置，使用400目的滤袋进行过滤，滤饼作为醇洗过的湿魔芋粉用于寡糖的制备原料；
(2)酶法水解魔芋粉制备高浓度魔芋寡糖液
取60℃热水加入甘露聚糖酶，形成甘露聚糖酶活力达到40-50IU/mL的酶溶液，加入到酒精洗过的魔芋精粉中，边加边搅拌混合，水解反应30-50min，控制水解程度，维持平均聚合度DP在10-15，形成的魔芋寡糖的各组分中，二糖-六糖的成分含量控制在30-40％比例，六糖-十八糖的成分含量维持在60％；将酶解液加热到80℃，保温20min，进行甘露聚糖酶的灭活；
采用60目筛网过滤进行预处理，对过滤液在8000-10000rpm条件下进行离心，取上清液通过10000Da的超滤膜进行寡糖的筛分，获得寡糖分子量在180-3000Da的混合物；再通过分子量为200Da的纳滤膜进行过滤浓缩，形成分子量为342-3000Da混合寡糖，通过离心喷雾干燥设备进行干燥，收集魔芋寡糖粉；
(3)制备魔芋寡糖胶
取魔芋寡糖粉加入40-50％的酒精，搅拌混合，添加10％的甘油，放入到微波炉中，微波强度100％，加热至溶液处于沸腾状态，悬浮液溶解后变澄清，继续加热，通过微波加热处理使魔芋寡糖浓度达到70-80％，形成的寡糖混合物的粘性维持在2000-6000mPa.S的高粘性特征，即形成魔芋寡糖胶，用于涂抹手臂、皮肤表面，迅速形成干燥薄膜，对外来病原形成屏障保护作用。
与现有技术相比本发明的有益效果是：
1)魔芋精粉通过甘露聚糖酶适当处理后，形成一定浓度的小分子生物活性寡糖，该产物具有一定的粘性和亲水性特效性，涂抹能形成一层薄膜型的寡糖膜，具有保护伤口愈合，防护外来感染物的保护屏障作用。
2)采用酶法水解制备获得的高浓度的魔芋寡糖，控制水解程度，维持平均聚合度DP在10-15，形成的寡糖的粘度和吸湿性较好。
3)采用微波法处理酒精溶解的高浓度魔芋寡糖形成的不溶颗粒，通过高温溶解，结合酒精和玫瑰花香的酯内物质的分子之间的结合，快速脱水，浓缩形成多糖链复合物。形成的寡糖浓度在70-80％(W/V)，在涂抹皮肤上形成涂层，能迅速在2min下凝固成型，有利于伤口患处的修复和阻断外来污染源。
4)利用寡糖的生物活性特点及吸湿性，通过微波加热处理进行分子组装，获得寡糖胶产品。相对于采用直接加热法浓缩处理，形成的产品在抑制微生物方面较弱，形成的产品浑浊的缺陷，本发明采用微波法，有利于小分子糖的聚合和高浓度物质的溶解，增强了产品的生物活性。
附图说明
图1是本发明一实施例的流程图。
具体实施方式
下面结合附图所示的各实施方式对本发明进行详细说明，但应当说明的是，这些实施方式并非对本发明的限制，本领域普通技术人员根据这些实施方式所作的功能、方法、或者结构上的等效变换或替代，均属于本发明的保护范围之内。
参图1所示，本实施例提供了一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法，包括：
(1)去除魔芋粉中的生物碱
取魔芋精粉100g，按照料水比1：2加入80％的酒精200mL进行醇洗，搅拌混合30min，静置30min，使用400目的滤袋进行过滤，滤饼作为醇洗过的湿魔芋粉用于寡糖的制备原料；
(2)酶法水解魔芋粉制备高浓度魔芋寡糖液
取60℃热水300mL，加入甘露聚糖酶，使溶液的甘露聚糖酶活力达到40-50IU/mL的酶溶液，加入到酒精洗过的湿魔芋精粉中，边加边搅拌混合，水解反应30-50min，控制水解程度，维持平均聚合度DP在10-15。在该条件下，形成的魔芋寡糖的各组分中，二糖-六糖的成分含量能控制在30-40％比例，六糖—十八糖的成分含量维持在60％。对酶解液加热到80℃，保温20min，进行甘露聚糖酶的灭活；采用60目筛网过滤进行预处理，对过滤液在8000-10000rpm条件下进行离心，取上清液通过10000Da的超滤膜进行寡糖的筛分，获得寡糖分子量在180-3000Da的混合物；再通过分子量为200Da的纳滤膜进行过滤浓缩，形成分子量为342-3000Da混合寡糖，通过离心喷雾干燥设备进行干燥，收集魔芋寡糖粉；
(3)制备魔芋寡糖胶
魔芋寡糖具有的生物活性特征，而且由于魔芋寡糖的吸潮特性和产品的的保湿性要求，本实施案例采用40-50％的酒精溶液进行溶解混合，添加10％的甘油，起到杀菌、保湿的作用。称取魔芋寡糖粉25g，加入40-50％的酒精15mL，搅拌混合，放入到微波炉中，微波强度100％，加热至溶液处于沸腾状态，悬浮液溶解后变澄清，继续加热，通过微波加热处理使魔芋寡糖浓度达到70-80％，形成的寡糖混合物的粘性维持在2000-6000mPa.S的高粘性特征，即形成魔芋寡糖胶，用于涂抹手臂、皮肤表面，迅速形成干燥薄膜，对外来病原形成屏障保护作用。
该利用魔芋寡糖复膜特性制备伤口愈合剂的方法具有如下技术效果：
魔芋寡糖大分子物质为主，具有一定的粘性特征；通过酒精和甘油的混合，通过微波处理形成一定特殊的纳米材料，具有快速干燥，吸附作用。
魔芋的水解时间短，形成的寡糖偏向于大分子部分，聚合度DP在10-15。该魔芋寡糖胶通过微波处理后具有高粘性，溶解性好，微波处理后杀菌彻底，通过酒精溶解处理后具有抑菌效应。
在应用方面由于魔芋寡糖作为生物活性物质，能修复人体细胞，抑制病原微生物的生长，涂抹细胞表面能覆膜成型，细胞包埋病原菌，抑制生长，用于人体表面皮肤的保护和抗菌作用、伤口的愈合。
对于本领域技术人员而言，显然本发明不限于上述示范性实施例的细节，而且在不背离本发明的精神或基本特征的情况下，能够以其他的具体形式实现本发明。因此，无论从哪一点来看，均应将实施例看作是示范性的，而且是非限制性的，本发明的范围由所附权利要求而不是上述说明限定，因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。
Claims (1)
1.一种利用魔芋寡糖复膜特性制备伤口愈合剂的方法，其特征在于，包括如下步骤：
(1)去除魔芋粉中的生物碱
取魔芋粉，按照料水比1：2加入80％的酒精进行醇洗，搅拌混合，静置，使用400目的滤袋进行过滤，滤饼作为醇洗过的湿魔芋粉用于寡糖的制备原料；
(2)酶法水解魔芋粉制备高浓度魔芋寡糖液
取60℃热水加入甘露聚糖酶，形成甘露聚糖酶活力达到40-50IU/mL的酶溶液，加入到酒精洗过的魔芋精粉中，边加边搅拌混合，水解反应30-50min，控制水解程度，维持平均聚合度DP在10-15，形成的魔芋寡糖的各组分中，二糖-六糖的成分含量控制在30-40％比例，六糖-十八糖的成分含量维持在60％；将酶解液加热到80℃，保温20min，进行甘露聚糖酶的灭活；
采用60目筛网过滤进行预处理，对过滤液在8000-10000rpm条件下进行离心，取上清液通过10000Da的超滤膜进行寡糖的筛分，获得寡糖分子量在180-3000Da的混合物；再通过分子量为200Da的纳滤膜进行过滤浓缩，形成分子量为342-3000Da混合寡糖，通过离心喷雾干燥设备进行干燥，收集魔芋寡糖粉；
(3)制备魔芋寡糖胶
取魔芋寡糖粉加入40-50％的酒精，搅拌混合，添加10％的甘油，放入到微波炉中，微波强度100％，加热至溶液处于沸腾状态，悬浮液溶解后变澄清，继续加热，通过微波加热处理使魔芋寡糖浓度达到70-80％，形成的寡糖混合物的粘性维持在2000-6000mPa.S的高粘性特征，即形成魔芋寡糖胶，用于涂抹手臂、皮肤表面，迅速形成干燥薄膜，对外来病原形成屏障保护作用。
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