CN112940082A - 一种抗菌肽及其应用 - Google Patents
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Abstract
本发明公开了一种抗菌肽及其应用，该抗菌肽为具有如SEQ ID NO.1所示的氨基酸序列的抗菌肽YHX‑1。该抗菌肽具有广谱的抗菌活性，能够抑制单核细胞增生李斯特氏菌、金黄色葡萄球菌、变异链球菌等革兰氏阳性菌和大肠杆菌、沙门氏菌等革兰氏阴性菌，且均具有较高的抗菌活性。另外抗菌肽的溶血活性较低，毒性较小，而且合成序列短、分子量小，化学合成难度小，在能够良好特异性地杀死生物体内的病原菌同时保证了安全性，此外，还能节约规模化生产成本。
Description
技术领域
本发明涉及抗菌剂，具体地，涉及一种抗菌肽及其应用。
背景技术
自世界上第一种抗生素—青霉素问世以来，抗生素就在人类与病原体的斗争中一直发挥着不可替代的作用。但近年来抗生素滥用导致耐药菌株不断增多及多重耐药菌的出现，对治疗临床感染构成了严重的威胁。如耐甲氧西林金黄葡萄球菌(Methicillin-resistantStaphylococcusaureus，MRSA)，目前已具备了广谱耐药性，对所有与甲氧西林结构相近的β-内酰胺类和头孢类抗生素，以及氨基糖苷类、大环内酯类、四环素类、氟喹诺酮类、磺胺类、利福平均产生不同程度的耐药。MRSA同时也是医院院内感染的一种常见病原菌，其感染多见于外伤与手术，2016年，国内主要地区医院MRSA的平均检出率为38.4％，部分医院的检出率高达53.5％-75.3％，严重影响到了病人的康复过程。因此解决病原菌的耐药性问题已经刻不容缓。而开发新型抗菌药物是解决此问题的一个重要途径。其中抗菌肽凭借独特的穿膜机制以及快速的杀菌效率，不易产生耐药性，成为抗生素最有力的替代者。
抗菌肽具有包括抗细菌、抗真菌、抗病毒、杀灭寄生虫、抑制肿瘤以及调节免疫系统的多种生物活性。同时，其独特的膜破坏机制和胞内杀菌机制不易产生耐药性，因而具有广泛的应用前景。
截止到2019年5月，权威的抗菌肽数据库APD3(http://aps.unmc.edu/AP/main.php)已经收录了3167条抗菌肽序列。然而，目前大部分抗菌肽还存在生产成本高、生物活性较低等问题。因此，通过理性设计开发出结构简单、生物活性高的抗菌肽是抗菌肽研究开发领域的当务之急。
发明内容
针对以上问题，本发明提供了一种抗菌肽，该抗菌肽合成序列短、溶血率低、生物活性高，能够特异性地杀死生物体内的病原菌。
为了实现上述目的，本发明一方面提供一种抗菌肽，所述抗菌肽为具有如SEQ IDNO.1所示的氨基酸序列的抗菌肽YHX-1。
具体地，所述抗菌肽YHX-1的分子量为1481.97Da，净电荷数为+5。
本发明另一方面提供了上述抗菌肽在制备治疗单核细胞增生李斯特氏菌、变异链球菌、沙门氏菌、大肠杆菌或金黄色葡萄球菌感染性疾病药物中的应用。
具体地，所述抗菌肽YHX-1对单核细胞增生李斯特氏菌的最小抑菌浓度为4μg/mL；
所述抗菌肽YHX-1对变异链球菌的最小抑菌浓度为16μg/mL；
所述抗菌肽YHX-1对沙门氏菌的最小抑菌浓度为4μg/mL；
所述抗菌肽YHX-1对大肠杆菌的最小抑菌浓度为32μg/mL；
所述抗菌肽YHX-1对金黄色葡萄球菌的最小抑菌浓度为128μg/mL。
本发明第三方面提供一种生物抗菌剂，该生物抗菌剂包括上述的抗菌肽YHX-1。
本发明第四方面提供一种动物饲料，该动物饲料包括上述的抗菌肽YHX-1。
本发明第五方面提供一种防腐剂，该防腐剂包括上述的抗菌肽YHX-1。
具体地，所述防腐剂为用于食品的防腐剂。
具体地，所述防腐剂为用于化妆品的防腐剂。
本发明第六方面提供一种洗涤组合物，该洗涤组合物包括上述的抗菌肽YHX-1。
通过上述技术方案，本发明实现了以下有益效果：
1、本发明的抗菌肽具有广谱的抗菌活性，能够抑制单核细胞增生李斯特氏菌、金黄色葡萄球菌、变异链球菌等革兰氏阳性菌和大肠杆菌、沙门氏菌等革兰氏阴性菌，且均具有较高的抗菌活性。
2、本发明所述的抗菌肽的溶血活性较低，且合成序列短、分子量小，化学合成难度小，在能够良好特异性地杀死生物体内的病原菌同时节约了规模化生产成本。
附图说明
图1为本发明提供的抗菌肽YHX-1的二级结构预测示意图；
图2为本发明提供的抗菌肽YHX-1在333μg/mL浓度条件下对单核细胞增生李斯特氏菌的抑菌效果示意图；
图3为本发明提供的抗菌肽YHX-1在333μg/mL浓度条件下对大肠杆菌的抑菌效果示意图；
图4为本发明提供的抗菌肽YHX-1在333μg/mL浓度条件下对变异链球菌的抑菌效果示意图；
图5为本发明提供的抗菌肽YHX-1在333μg/mL浓度条件下对沙门氏菌的抑菌效果示意图。
具体实施方式
下面将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他实施例，都属于本发明保护的范围。
实施例1抗菌肽的设计
在抗菌肽从头设计及对构效关系理解的基础上，从ADP3数据库中筛选得到的对革兰氏阳性菌和革兰氏阴性菌有抑菌作用的843条抗菌肽序列进行序列参数分析(包括序列长度、带电荷数、疏水氨基酸比例和氨基酸组成)，之后按照出现频率最大的选取原则并结合理性设计思路，确定了新抗菌肽的各项序列参数，具体如下：
表1从头设计抗菌肽的序列参数
为了降低合成成本，同时降低细胞毒性，序列长度选择频率第二位的13。选取带正电氨基酸Lys和极性不带电荷氨基酸Ser组成抗菌肽的极性面，选取疏水性氨基酸Leu形成疏水面，来保证抗菌肽的两亲性结构。带电荷数确定为+5，保证抗菌肽对细菌细胞膜的亲和力。
采用生物信息学预测及分析工具，对不同氨基酸排列顺序的肽序列分子量、平均疏水值、平均疏水力矩、不稳定性指数、脂肪族指数、半衰期、螺旋轮图、成为AMPs(antimicrobial peptides，抗菌肽)的可能性以及二级结构进行预测，选出最优的抗菌肽序列。所用生物信息学工具如下：
物化性质分析网站：Expasy ProtParam(https://web.expasy.org/protparam/)
螺旋轮图及疏水性分析网站：Heliquest(https://heliquest.ipmc.cnrs.fr/)
AMPs可能性预测网站：CAMPR3(http://www.camp.bicnirrh.res.in/)
二级结构预测网站：Pep-fold(https://bioserv.rpbs.univ-paris-diderot.fr/services/PEP-FOLD3/)；
Zhang Lab(https://zhanglab.ccmb.med.umich.edu/)
通过上述工具得到的最优抗菌肽序列及物化性质如表2所示，二级结构预测结果如图1所示。
表2抗菌肽YHX-1的序列及物化性质
该抗菌肽YHX-1委托强耀生物科技(上海)有限公司合成。
实施例2抑菌活性
将金黄色葡萄球菌、大肠杆菌和沙门氏菌分别划线接种于LB固体培养基(Luria-Bertani培养基)，将单核细胞增生李斯特氏菌和变异链球菌划线接种于BHI固体培养基(脑心浸液琼脂培养基)，置于37℃恒温培养箱培养18h，挑取各个菌株的单菌落置于各自对应的液体培养基中，37℃恒温振荡培养12h。测量菌液的OD600值(溶液在600nm波长处的吸光值)，并将其稀释至1×106CFU/mL。
①抑菌圈实验
配置LB和BHI半固体培养基(琼脂质量分数0.6％)，每皿20mL加入7μL菌液振荡混匀后倒入摆放好牛津杯的培养皿中，待培养基冷却凝固后拔除牛津杯完成打孔。每孔中加入160μL抗菌肽溶液(配置使用1mg抗菌肽溶于3mL超纯水)。置于37℃恒温培养箱培养24h后测量抑菌圈直径，测试结果如图2-图5所示。
②最小抑菌浓度(MIC)的测定
向96孔板中加入培养至对数生长期的各株菌菌液(单核细胞增生李斯特氏菌、变异链球菌、金黄色葡萄球菌、沙门氏菌、大肠杆菌)50μL，同时在各孔中加入各浓度梯度抗菌肽溶液(512μg/mL、256μg/mL、128μg/mL、64μg/mL、32μg/mL、16μg/mL、8μg/mL、4μg/mL)50μL，并分别做阳性对照和阴性对照组。在37℃恒温培养箱培养8h后测量各孔的OD600值，取菌株生长被完全抑制的最低浓度作为抗菌肽的最小抑菌浓度(MIC)。
表3抗菌肽YHX-1的抑菌活性分析结果
从图2-图5可以看到具有明显的抑菌圈出现，结合表3，表明本发明的抗菌肽对单核细胞增生李斯特氏菌、变异链球菌、大肠杆菌、金黄色葡萄球菌和沙门氏菌具有抑制作用，对单核细胞增生李斯特氏菌的最小抑菌浓度为4μg/mL，对变异链球菌的的最小抑菌浓度为16μg/mL，对大肠杆菌的最小抑菌浓度为32μg/mL，对金黄色葡萄球菌的最小抑菌浓度为128μg/mL，对沙门氏菌的最小抑菌浓度为4μg/mL，可见，本发明的抗菌肽对上述菌尤其是单核细胞增生李斯特氏菌、变异链球菌、沙门氏菌等具有优异的抑制作用。
实施例3溶血活性
取健康兔血液1mL加于肝素抗凝管中，在1000xg转速下离心10min后取沉淀，用PBS缓冲液冲洗3次，将红细胞用10mL PBS重悬。用PBS缓冲液将抗菌肽YHX-1的浓度调整至4μg/mL、8μg/mL、16μg/mL、32μg/mL、128μg/mL、256μg/mL、512μg/mL，加入等体积的红细胞悬液。以PBS缓冲液作为阴性对照，以0.1％Tritonx-100(聚乙二醇辛基苯基醚)作为阴性对照，在37℃恒温培养1h后取出，1000xg离心10min，取出上清液用酶标仪在570nm处测OD值。
溶血率计算公式为：溶血率＝(AT-A0)/(AC-A0)×100％。
式中：AT为实验组的吸光值，AC为阳性对照组的吸光值，A0为阴性对照组的吸光值。
表4抗菌肽YHX-1的溶血活性(测定3次取平均值)
由表4可知，在最小抑菌浓度(MIC条件下)，抗菌肽YHX-1的溶血率均低于10％，说明抗菌肽YHX-1有较好的安全性，在制作生物抗菌剂等领域的应用前景较大，可进一步深入研究和开发利用。
序列表
<110> 扬州市扬大康源乳业有限公司
<120> 一种抗菌肽及其应用
<160> 1
<170> SIPOSequenceListing 1.0
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<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
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Gly Lys Leu Leu Ser Lys Leu Leu Lys Lys Leu Leu Lys
1 5 10
Claims (10)
1.一种抗菌肽，其特征在于，该抗菌肽为具有如SEQ ID NO.1所示的氨基酸序列的抗菌肽YHX-1。
2.根据权利要求1所述的抗菌肽，其特征在于，所述抗菌肽YHX-1的分子量为1481.97Da，净电荷数为+5。
3.根据权利要求1至2中任一项所述的抗菌肽在制备治疗单核细胞增生李斯特氏菌、变异链球菌、沙门氏菌、大肠杆菌或金黄色葡萄球菌感染性疾病药物中的应用。
4.根据权利要求3所述的应用，其特征在于，所述抗菌肽YHX-1对单核细胞增生李斯特氏菌的最小抑菌浓度为4μg/mL；
所述抗菌肽YHX-1对变异链球菌的最小抑菌浓度为16μg/mL；
所述抗菌肽YHX-1对沙门氏菌的最小抑菌浓度为4μg/mL；
所述抗菌肽YHX-1对大肠杆菌的最小抑菌浓度为32μg/mL；
所述抗菌肽YHX-1对金黄色葡萄球菌的最小抑菌浓度为128μg/mL。
5.一种生物抗菌剂，其特征在于，包括根据权利要求1至2中任一项所述的抗菌肽YHX-1。
6.一种动物饲料，其特征在于，包括根据权利要求1至2中任一项所述的抗菌肽YHX-1。
7.一种防腐剂，其特征在于，包括根据权利要求1至2中任一项所述的抗菌肽YHX-1。
8.根据权利要求7所述的防腐剂，其特征在于，所述防腐剂为用于食品的防腐剂。
9.根据权利要求7所述的防腐剂，其特征在于，所述防腐剂为用于化妆品的防腐剂。
10.一种洗涤组合物，其特征在于，包括根据权利要求1至2中任一项所述的抗菌肽YHX-1。
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113336827A (zh) * | 2021-06-30 | 2021-09-03 | 中国海洋大学 | 一种低分子量抗菌肽yhx-2及其组合物和应用 |
| CN113754750A (zh) * | 2021-09-30 | 2021-12-07 | 华中农业大学 | 一种抗菌肽及其在水产养殖中的应用 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20190033836A (ko) * | 2017-09-22 | 2019-04-01 | 조선대학교산학협력단 | 홍어 껍질 유래의 신규한 항균 펩타이드 및 이의 용도 |
| US20200040035A1 (en) * | 2016-06-23 | 2020-02-06 | Konkuk University Industrial Cooperation Corp | Antimicrobial peptide having synergistic antibacterial effect with antibiotics on multidrug resistant bacteria, and use thereof |
| CN111925430A (zh) * | 2020-08-18 | 2020-11-13 | 中国海洋大学 | 抗菌肽及其应用 |
| CN111944020A (zh) * | 2020-08-18 | 2020-11-17 | 中国海洋大学 | 抗菌肽及其应用 |
-
2021
- 2021-04-23 CN CN202110442926.6A patent/CN112940082B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20200040035A1 (en) * | 2016-06-23 | 2020-02-06 | Konkuk University Industrial Cooperation Corp | Antimicrobial peptide having synergistic antibacterial effect with antibiotics on multidrug resistant bacteria, and use thereof |
| KR20190033836A (ko) * | 2017-09-22 | 2019-04-01 | 조선대학교산학협력단 | 홍어 껍질 유래의 신규한 항균 펩타이드 및 이의 용도 |
| CN111925430A (zh) * | 2020-08-18 | 2020-11-13 | 中国海洋大学 | 抗菌肽及其应用 |
| CN111944020A (zh) * | 2020-08-18 | 2020-11-17 | 中国海洋大学 | 抗菌肽及其应用 |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN115246878B (zh) * | 2021-03-31 | 2024-02-13 | 禾美生物科技(浙江)有限公司 | 一种抗菌肽及其在化妆品中的用途 |
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| CN113336827A (zh) * | 2021-06-30 | 2021-09-03 | 中国海洋大学 | 一种低分子量抗菌肽yhx-2及其组合物和应用 |
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