CN112715547A - 一种壳聚糖-溶菌酶复合抑菌剂及其制备方法与应用 - Google Patents

一种壳聚糖-溶菌酶复合抑菌剂及其制备方法与应用 Download PDF

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CN112715547A
CN112715547A CN202011616498.6A CN202011616498A CN112715547A CN 112715547 A CN112715547 A CN 112715547A CN 202011616498 A CN202011616498 A CN 202011616498A CN 112715547 A CN112715547 A CN 112715547A
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chitosan
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曹逊
李泽平
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Nanjing Qiyou Biological Technology Co ltd
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Abstract

本发明公开了一种壳聚糖‑溶菌酶复合抑菌剂及其制备方法与应用。该抑菌剂包括以下按重量份数计的组分：柠檬酸3‑6份、溶菌酶12‑18份、纳米银1‑3份、纳米铜5‑8份、乙醇8‑18份、丁香酚1‑2份、发泡剂1‑3份、尼泊金复合酯钠2‑5份、戊二醇10‑15份、香茅草精油1‑3份、水60‑80份。而混合物A、混合物B等与香茅草精油混合后，浓缩，使得抑菌剂的外围包裹这一层油膜，通过隔绝的方式破坏了细菌的生成环境，从而达到了高的抑菌效果，且组分简单，易于制备，适合产业化。

Description

一种壳聚糖-溶菌酶复合抑菌剂及其制备方法与应用
技术领域
本发明涉及抑菌剂制备技术领域，具体涉及一种壳聚糖-溶菌酶复合抑菌剂及其制备方法与应用。
背景技术
生物抑菌物质具有高效无毒、来源广泛等特点，在食品保藏、防腐、疾病治疗等方面逐渐引起人们的重视。常见的生物抑菌剂包括乳酸链球菌素、壳聚糖、溶菌酶、大蒜素等。
溶菌酶主要通过破坏细胞壁中的N-乙酰胞壁酸和N-乙酰氨基葡萄糖之间的β-1，4糖苷键，使细胞壁不溶性黏多糖分解成可溶性糖肽，导致细胞壁破裂内容物逸出而使细胞溶解。但是溶菌酶在体外进化催化反应时，其稳定性与活力较差。
壳聚糖是天然多糖甲壳素脱除乙酰基的产物，具有生物降解性、生物相容性、无毒性等多种生理功能，广泛应用于食品添加剂、纺织、农业、环保、化妆品等领域。壳聚糖是一种长链状高分子材料，可以缠绕在溶菌酶酶分子的表面，增强溶菌酶的热稳定性、pH稳定性以及贮存稳定性等。同时，壳聚糖对大肠杆菌、荧光假单胞菌、金黄色葡萄球菌、枯草杆菌等也具有良好的抑制作用。壳聚糖的衍生物也有很好地抗菌性。
如何提高溶菌酶在体外的活性，并有效地与溶菌酶协同作用，提高抑菌剂的抑菌效果，是值得探究的课题。
发明内容
针对现有技术的不足，本发明提供了一种壳聚糖-溶菌酶复合抑菌剂及其制备方法与应用，所得壳聚糖-溶菌酶复合抑菌剂成分简单，安全有效，且制备方法简单，适合产业化。
一种壳聚糖-溶菌酶复合抑菌剂，包括以下按重量份数计的组分：柠檬酸3-6份、溶菌酶12-18份、纳米银1-3份、纳米铜5-8份、乙醇8-18份、丁香酚1-2份、发泡剂1-3份、尼泊金复合酯钠2-5份、戊二醇10-15份、香茅草精油1-3份、水60-80份。
作为改进的是，上述壳聚糖-溶菌酶复合抑菌剂，包括以下按重量份数计的组分：柠檬酸5份、溶菌酶15份、纳米银2份、纳米铜6份、乙醇12份、丁香酚1.5份、发泡剂2份、尼泊金复合酯钠3份、戊二醇12份、香茅草精油2份、水75份。
作为改进的是，所述溶菌酶为蛋清溶菌酶。
上述壳聚糖-溶菌酶复合抑菌剂的制备方法，包括以下步骤：
步骤1，将纳米银、纳米铜、尼泊金复合酯钠混合后，投入高速剪切机中剪切1-3分得混合物A；
步骤2，将柠檬酸、乙醇、丁香酚、发泡剂、戊二醇混合，投入乳化机中乳化1-3分钟得乳化混合物B；
步骤3，将混合物A、混合物B、溶菌酶以及香茅草精油混合，并加入水，高速搅拌后，浓缩至含水率为25-30%的复合抑菌剂。
作为改进的是，步骤1中高速剪切机的剪切速度为1500-2000rpm。
作为改进的是，步骤3中高速搅拌的速度为300-400rpm。
上述壳聚糖-溶菌酶复合抑菌剂在制备洗手液或空气消毒剂上的应用。
壳聚糖是一种长链状高分子材料，可以缠绕在溶菌酶酶分子的表面，增强溶菌酶的热稳定性、pH稳定性以及贮存稳定性等。同时，壳聚糖对大肠杆菌、荧光假单胞菌、金黄色葡萄球菌、枯草杆菌等也具有良好的抑制作用。壳聚糖的衍生物也有很好地抗菌性。壳聚糖-溶菌酶复合抑菌制剂具有更好的抑菌效果。
有益效果：
与现有技术相比，本发明一种壳聚糖-溶菌酶复合抑菌剂及其制备方法与应用，将化学抑菌和生物抑菌相糅合的方法制备新型的复合抑菌剂。纳米银和溶菌酶通过破坏细菌壁的方式影响细菌的生长，而混合物A、混合物B等与香茅草精油混合后，浓缩，使得抑菌剂的外围包裹这一层油膜，通过隔绝的方式破坏了细菌的生成环境，从而达到了高的抑菌效果，且组分简单，易于制备，适合产业化。
具体实施方案
实施例1
一种壳聚糖-溶菌酶复合抑菌剂，包括以下按重量份数计的组分：柠檬酸3-6份、蛋清溶菌酶12-18份、纳米银1-3份、纳米铜5-8份、乙醇8-18份、丁香酚1-2份、发泡剂1-3份、尼泊金复合酯钠2-5份、戊二醇10-15份、香茅草精油1-3份、水60-80份。
上述壳聚糖-溶菌酶复合抑菌剂的制备方法，包括以下步骤：
步骤1，将纳米银、纳米铜、尼泊金复合酯钠混合后，投入高速剪切机中1500rpm剪切1-3分得混合物A；
步骤2，将柠檬酸、乙醇、丁香酚、发泡剂、戊二醇混合，投入乳化机中乳化1-3分钟得乳化混合物B；
步骤3，将混合物A、混合物B、溶菌酶以及香茅草精油混合，并加入水，300rpm高速搅拌后，浓缩至含水率为25-30%的复合抑菌剂。
将上述100ml复合抑菌剂加入到200μl的菌浓度为106CFU/ml的大肠杆菌ATCC25922菌液和200μl的LB培养基中作用两个小时，对大肠杆菌ATCC25922的抑菌率为104.12。
实施例2
一种壳聚糖-溶菌酶复合抑菌剂，包括以下按重量份数计的组分：柠檬酸5份、蛋清溶菌酶15份、纳米银2份、纳米铜6份、乙醇12份、丁香酚1.5份、发泡剂2份、尼泊金复合酯钠3份、戊二醇12份、香茅草精油2份、水75份。
上述壳聚糖-溶菌酶复合抑菌剂的制备方法，包括以下步骤：
步骤1，将纳米银、纳米铜、尼泊金复合酯钠混合后，投入高速剪切机中1800rpm剪切1-3分得混合物A；
步骤2，将柠檬酸、乙醇、丁香酚、发泡剂、戊二醇混合，投入乳化机中乳化1-3分钟得乳化混合物B；
步骤3，将混合物A、混合物B、溶菌酶以及香茅草精油混合，并加入水，380rpm高速搅拌后，浓缩至含水率为25-30%的复合抑菌剂。
将上述100ml复合抑菌剂加入到200μl的菌浓度为106CFU/ml的大肠杆菌ATCC25922菌液和200μl的LB培养基中作用两个小时，对大肠杆菌ATCC25922的抑菌率为104.65。
实施例3
一种壳聚糖-溶菌酶复合抑菌剂，包括以下按重量份数计的组分：柠檬酸3-6份、蛋清溶菌酶12-18份、纳米银1-3份、纳米铜5-8份、乙醇8-18份、丁香酚1-2份、发泡剂1-3份、尼泊金复合酯钠2-5份、戊二醇10-15份、香茅草精油1-3份、水60-80份。
上述壳聚糖-溶菌酶复合抑菌剂的制备方法，包括以下步骤：
步骤1，将纳米银、纳米铜、尼泊金复合酯钠混合后，投入高速剪切机中2000rpm剪切1-3分得混合物A；
步骤2，将柠檬酸、乙醇、丁香酚、发泡剂、戊二醇混合，投入乳化机中乳化1-3分钟得乳化混合物B；
步骤3，将混合物A、混合物B、溶菌酶以及香茅草精油混合，并加入水，400rpm高速搅拌后，浓缩至含水率为25-30%的复合抑菌剂。
将上述100ml复合抑菌剂加入到200μl的菌浓度为106CFU/ml的大肠杆菌ATCC25922菌液和200μl的LB培养基中作用两个小时，对大肠杆菌ATCC25922的抑菌率为103.85。
对比例1
除不含有纳米银外，其余同实施例2。
将上述100ml复合抑菌剂加入到200μl的菌浓度为106CFU/ml的大肠杆菌ATCC25922菌液和200μl的LB培养基中作用两个小时，对大肠杆菌ATCC25922的抑菌率为101.25。明显低于本发明的抑菌效果。
对比例2
除不含香茅草精油外，其余同实施例2。
将上述100ml复合抑菌剂加入到200μl的菌浓度为106CFU/ml的大肠杆菌ATCC25922菌液和200μl的LB培养基中作用两个小时，对大肠杆菌ATCC25922的抑菌率为101.35，明显低于本发明的抑菌效果。
对比例3
除不含有溶菌酶外，其余同实施例2。
将上述100ml复合抑菌剂加入到200μl的菌浓度为106CFU/ml的大肠杆菌ATCC25922菌液和200μl的LB培养基中作用两个小时，对大肠杆菌ATCC25922的抑菌率为102.58，明显低于本发明的抑菌效果。
综上所述，本发明将化学抑菌和生物抑菌相糅合的方法制备新型的复合抑菌剂。纳米银和溶菌酶通过破坏细菌壁的方式影响细菌的生长，而混合物A、混合物B等与香茅草精油混合后，浓缩，使得抑菌剂的外围包裹这一层油膜，通过隔绝的方式破坏了细菌的生成环境，从而达到了高的抑菌效果，且组分简单，易于制备，适合产业化。
以上所述，仅为本发明较佳的具体实施方式，本发明的保护范围不限于此，任何熟悉本技术领域的技术人员在本发明披露的技术范围内，可显而易见地得到的技术方案的简单变化或等效替换均落入本发明的保护范围内。

Claims (7)

1.一种壳聚糖-溶菌酶复合抑菌剂，其特征在于，包括以下按重量份数计的组分：柠檬酸3-6份、溶菌酶12-18份、纳米银1-3份、纳米铜5-8份、乙醇8-18份、丁香酚1-2份、发泡剂1-3份、尼泊金复合酯钠2-5份、戊二醇10-15份、香茅草精油1-3份、水60-80份。
2.根据权利要求1所述的壳聚糖-溶菌酶复合抑菌剂，其特征在于，包括以下按重量份数计的组分：柠檬酸5份、溶菌酶15份、纳米银2份、纳米铜6份、乙醇12份、丁香酚1.5份、发泡剂2份、尼泊金复合酯钠3份、戊二醇12份、香茅草精油2份、水75份。
3.根据权利要求1所述的壳聚糖-溶菌酶复合抑菌剂，其特征在于，所述溶菌酶为蛋清溶菌酶。
4.基于权利要求1所述的壳聚糖-溶菌酶复合抑菌剂的制备方法，包括以下步骤：
步骤1，将纳米银、纳米铜、尼泊金复合酯钠混合后，投入高速剪切机中剪切1-3分得混合物A；
步骤2，将柠檬酸、乙醇、丁香酚、发泡剂、戊二醇混合，投入乳化机中乳化1-3分钟得乳化混合物B；
步骤3，将混合物A、混合物B、溶菌酶以及香茅草精油混合，并加入水，高速搅拌后，浓缩至含水率为25-30%的复合抑菌剂。
5.根据权利要求4所述的壳聚糖-溶菌酶复合抑菌剂的制备方法，其特征在于，步骤1中高速剪切机的剪切速度为1500-2000rpm。
6.根据权利要求4所述的壳聚糖-溶菌酶复合抑菌剂的制备方法，其特征在于，步骤3中高速搅拌的速度为300-400rpm。
7.基于权利要求1所述的壳聚糖-溶菌酶复合抑菌剂在制备洗手液或空气消毒剂上的应用。
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