CN114149483A - 一种抗菌肽组合物及其应用 - Google Patents
一种抗菌肽组合物及其应用 Download PDFInfo
- Publication number
- CN114149483A CN114149483A CN202110590640.2A CN202110590640A CN114149483A CN 114149483 A CN114149483 A CN 114149483A CN 202110590640 A CN202110590640 A CN 202110590640A CN 114149483 A CN114149483 A CN 114149483A
- Authority
- CN
- China
- Prior art keywords
- mamp
- gram
- asp
- leu
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims abstract description 47
- 239000000203 mixture Substances 0.000 title claims abstract description 31
- MYWUZJCMWCOHBA-UHFFFAOYSA-N n-methyl-1-phenylpropan-2-amine Chemical compound CNC(C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-UHFFFAOYSA-N 0.000 claims abstract description 42
- 241000894006 Bacteria Species 0.000 claims abstract description 34
- 102000044503 Antimicrobial Peptides Human genes 0.000 claims abstract description 30
- 108700042778 Antimicrobial Peptides Proteins 0.000 claims abstract description 30
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 16
- 241000233866 Fungi Species 0.000 claims abstract description 14
- 241000605862 Porphyromonas gingivalis Species 0.000 claims abstract description 12
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 12
- 241000194019 Streptococcus mutans Species 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims abstract description 12
- 241000222122 Candida albicans Species 0.000 claims abstract description 11
- 229940095731 candida albicans Drugs 0.000 claims abstract description 11
- 244000005700 microbiome Species 0.000 claims abstract description 11
- 241000588724 Escherichia coli Species 0.000 claims abstract description 10
- 241000192125 Firmicutes Species 0.000 claims abstract description 9
- 102000014171 Milk Proteins Human genes 0.000 claims abstract description 6
- 108010011756 Milk Proteins Proteins 0.000 claims abstract description 6
- 229940079593 drug Drugs 0.000 claims abstract description 6
- 235000013305 food Nutrition 0.000 claims abstract description 6
- 235000021239 milk protein Nutrition 0.000 claims abstract description 6
- 239000004480 active ingredient Substances 0.000 claims abstract description 3
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 29
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 25
- 229920001184 polypeptide Polymers 0.000 claims description 16
- 230000000844 anti-bacterial effect Effects 0.000 claims description 12
- 239000003966 growth inhibitor Substances 0.000 claims description 7
- 230000000813 microbial effect Effects 0.000 claims description 4
- 239000000969 carrier Substances 0.000 claims description 3
- 208000017520 skin disease Diseases 0.000 claims description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 2
- 208000025157 Oral disease Diseases 0.000 claims description 2
- 230000002924 anti-infective effect Effects 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 208000015181 infectious disease Diseases 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 208000030194 mouth disease Diseases 0.000 claims description 2
- 239000002537 cosmetic Substances 0.000 claims 6
- 239000013588 oral product Substances 0.000 claims 5
- 229940023486 oral product Drugs 0.000 claims 3
- 229960005475 antiinfective agent Drugs 0.000 claims 2
- 239000004599 antimicrobial Substances 0.000 claims 2
- 210000000214 mouth Anatomy 0.000 claims 2
- 239000002671 adjuvant Substances 0.000 claims 1
- 201000010099 disease Diseases 0.000 claims 1
- 239000000047 product Substances 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 abstract description 17
- 239000002994 raw material Substances 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 description 12
- 239000007788 liquid Substances 0.000 description 11
- 230000003385 bacteriostatic effect Effects 0.000 description 9
- 230000001580 bacterial effect Effects 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 238000009631 Broth culture Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 208000035473 Communicable disease Diseases 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000006137 Luria-Bertani broth Substances 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 229940124350 antibacterial drug Drugs 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001646716 Escherichia coli K-12 Species 0.000 description 1
- 206010022678 Intestinal infections Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000007218 ym medium Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Dermatology (AREA)
- Biochemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Birds (AREA)
- Gastroenterology & Hepatology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明涉及一种抗菌肽组合物MAMP,该组合物由三个来自水解牛奶蛋白的抗菌肽MAMP‑01、MAMP‑02和MAMP‑03组成。MAMP‑01的氨基酸序列为Ala‑Ala‑Ser‑Asp‑Ile‑Ser‑Leu‑Leu(AASDISLL),MAMP‑02的氨基酸序列为Leu‑Asp‑Ala‑Gln‑Ser‑Asp‑Pro‑Leu‑Arg(LDAQSAPLR),MAMP‑03的氨基酸序列为Leu‑Asp‑Thr‑Asp‑Tyr‑Lys‑Lys‑Tyr(LDTDYKKY)。组合物MAMP对革兰氏阳性菌和革兰氏阴性菌、真菌具有广泛的抑制活性,包括大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变异链球菌和白色念珠菌等。具有原料天然安全和生产成本低的优点,可作为抑制微生物生长的活性成分应用于药物、食品、日用品中。
Description
技术领域
本发明涉及一种来源于水解牛奶蛋白的抗菌肽组合物MAMP及其应用。
背景技术
抗生素是能抵抗致病微生物的药物,是抗菌消炎药中最大的一类。抗生素是由细菌、真菌或其他微生物在生活过程中所产生的物质,具有抑制或杀灭细菌、真菌等致病微生物的作用,因此作为抗感染药物广泛地应用于各种感染性疾病。抗生素在治疗感染中的显著效果也导致了细菌耐药性,使部分感染性疾病的治疗变得愈加困难。抗菌肽作为一种新型的抗生素,具有广谱的抗菌活性,不会轻易使微生物产生耐药性,具有广阔的应用前景。
抗菌肽在自然界中广泛分布,作为被广泛关注的一种新型抗生素,越来越多的抗菌肽在微生物、动植物体内被发现。目前解析的抗菌肽构效关系中有两个较清晰的特征,分别是“带正电荷的阳离子”和“两亲性序列结构”。然而抗菌肽作为抗菌药物仍然面临许多待解决的问题:部分阳离子抗菌肽可以对哺乳动物的细胞产生较大的毒性作用;两亲性结构则要求氨基酸序列中有一定数量的碱性氨基酸,而碱性氨基酸易被蛋白酶水解的特性使得其作为药物进入人体后发挥效果大大受限。因此,开发安全、稳定、低细胞毒性的抗菌肽具有重要意义。
牛奶作为优质的天然食物,是丰富的蛋白质和多肽资源,兼顾抗菌肽的优势及优质安全的蛋白来源两个特质。牛奶中蛋白含量丰富,在蛋白酶的水解作用下,形成稳定存在的抗菌肽,作为微生物生长抑制剂具有良好的潜质。因此,从水解牛奶蛋白肽出发进一步寻找具有抑菌活性的肽会为健康安全的微生物生长抑制剂的制备提供新的方向。
发明内容
本发明的目的是提供一种来源于水解牛奶蛋白的抗菌肽组合物MAMP,在制备预防和/或降低由微生物引起的肠道感染疾病、口腔疾病、皮肤疾病的药物、日用品或者微生物生长抑制剂中的应用。
为实现上述目的,本发明以所述抗菌肽组合物MAMP为抑制微生物生长活性的有效成份。抗菌肽组合物MAMP由MAMP-01、MAMP-02和MAMP-03组成,具体为:
(1)MAMP-01为多肽AASDISLL,具有序列表SEQ ID NO:1中氨基酸序列;该多肽的氨基酸序列具体为:
Ala-Ala-Ser-Asp-Ile-Ser-Leu-Leu);分子量788.90Da,白色粉末状,易溶于水。
(2)MAMP-02为多肽LDAQSAPLR,具有序列表SEQ ID NO:2中氨基酸序列;该多肽的氨基酸序列具体为:
Leu-Asp-Ala-Gln-Ser-Asp-Pro-Leu-Arg;分子量970.09Da,白色粉末状,易溶于水。
(3)MAMP-03为多肽LDTDYKKY,具有序列表SEQ ID NO:3中氨基酸序列;该多肽的氨基酸序列具体为:
Leu-Asp-Thr-Asp-Tyr-Lys-Lys-Tyr;分子量1045.16Da,白色粉末状,易溶于水。
三个抗菌肽的比例为MAMP-01:MAMP-02:MAMP-03=1:0-10:5~20(重量比),优选MAMP-01:MAMP-02:MAMP-03=1:0-8:7~15(重量比),更优选MAMP-01:MAMP-02:MAMP-03=1:0-6:8~12(重量比)。
MAMP对大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变异链球菌、白色念珠菌的生长存在较强的抑制作用。MAMP可作为抗革兰氏阴性菌、革兰氏阳性菌、真菌的药物、食品、日用品的活性成份,其中可添加药学、食品、日用品上可接受的载体或辅料。
本发明与现有技术相比,具有如下有益效果:
本发明将来源于水解牛奶蛋白的抗菌肽按照其在酶解物中的含量进行组合,获得的抗菌肽组合物,与单一抗菌肽相比,组合物的抗菌效率提高、抑制微生物的种类更多。本发明首次验证了上述多肽组合物具有较好的抑菌活性,因此作为预防和/或降低由大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变异链球菌、白色念珠菌引起的感染疾病的药物和/或保健品和/或日用品或者微生物生长抑制剂具有良好的应用前景。
具体实施方式
实施例1
(1)MAMP的配置
于上海杰肽科技有限公司分别定制化学合成多肽MAMP-01(纯度为97.7%)、MAMP-02(纯度为95.4%)、MAMP-03(纯度为97.6%)。配置抗菌肽组合MAMP:1克MAMP-01+10克MAMP-02+20克MAMP-03。以革兰氏阴性菌中的大肠杆菌和牙龈卟啉单胞菌、革兰氏阳性菌中的金黄色葡萄球菌和变异链球菌、真菌白色念珠菌为靶标菌种,考察抗菌肽组合物MAMP的抗菌活性。
(2)菌株及复苏
大肠杆菌(Escherichia coli K12)、金黄色葡萄球菌(Staphylococcus aureus)、牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g.,ATCC33277)、变异链球菌(Streptococcus mutans,M.s.,ATCC25175)和白色念球菌(Candida albicans,ATCC10231)均购自中国菌种资源库保藏中心,以甘油保藏法保存于-80℃。实验前将大肠杆菌接种于LB肉汤培养基(北京索莱宝科技有限公司)中,将金黄色葡萄球菌接种于TSB肉汤培养基(北京索莱宝科技有限公司)中,将牙龈卟啉单胞菌和变异链球菌分别接种于BHI肉汤培养基(北京索莱宝科技有限公司)中,白色念球菌接种于YM培养基(青岛高科技工业园海博生物技术有限公司)中,靶标菌种均以1%体积量接种。于37℃条件下培养12h,将菌液以1%体积量再次接种至新鲜培养基中培养复苏。重复接种复苏操作2次,使菌株恢复活力。
(3)抗菌实验
分别取1ml培养过夜的菌液用PBS缓冲液洗涤2次后稀释成10^7菌液,获得大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变异链球菌和白色念球菌菌液。(MAMP、MAMP-01、MAMP-02、MAMP-03)样品分别溶解于上述菌培养对应的培养基中(大肠杆菌对应LB肉汤培养基,金黄色葡萄球菌对应TSB肉汤培养基,牙龈卟啉单胞菌和变异链球菌对应BHI肉汤培养基,白色念球菌对应YM培养基),配制成100mg/mL的抗菌肽溶液,过0.22um膜除菌备用,获得不同菌对应的肽溶液。将100ul不同菌对应的上述浓度肽溶液或菌培养对应的培养基分别与100ul上述对应的菌液混合(大肠杆菌对应LB肉汤培养基或其溶解样品后获得的菌液,金黄色葡萄球菌对应TSB肉汤培养基或其溶解样品后获得的菌液,牙龈卟啉单胞菌和变异链球菌对应BHI肉汤培养基或其溶解样品后获得的菌液,白色念球菌对应YM培养基或其溶解样品后获得的菌液),依次加入96孔细胞培养板的不同孔中,大肠杆菌、金黄色葡萄球菌、白色念珠菌于有氧环境37℃培养,牙龈卟啉单胞菌和变异链球菌于厌氧环境37℃培养。以上菌液分别培养12小时后于600nm处测吸光度。按照以下公式分别计算样品对应不同菌的抑菌率:
OD1对照组:菌液100ul+培养基100ul
OD2试验组:菌液100ul+肽溶液100ul
(4)实验结果
对五种菌的抑菌结果见表1,表中列出了抗菌肽组合物MAMP、MAMP-01、MAMP-02、MAMP-03的抑菌结果。结果表明抗菌肽组合物MAMP对革兰氏阴性、革兰氏阳性菌和真菌均有抑制活性,并且对于以上五种菌的抑制活性明显好于单一抗菌肽。
表1抗菌肽组合物MAMP的抑菌结果
实施例2
MAMP的配置:抗菌肽组合物MAMP:1克MAMP-01+5克MAMP-03。
其他实验方法与实施例1相同。
对五种菌的抑菌结果见表2,结果表明抗菌肽组合物MAMP对革兰氏阴性、革兰氏阳性菌和真菌均有抑制活性,并且对于以上五种菌的抑制活性明显好于单一抗菌肽。
表2抗菌肽组合物MAMP的抑菌结果
实施例3
MAMP的配置:抗菌肽组合物MAMP:1克MAMP-01+6克MAMP-02+15克MAMP-03。
其他实验方法与实施例1相同。
对五种菌的抑菌结果见表3,结果表明抗菌肽组合物MAMP对革兰氏阴性、革兰氏阳性菌和真菌均有抑制活性,并且对于以上五种菌的抑制活性明显好于单一抗菌肽。
表3抗菌肽组合物MAMP的抑菌结果
实施例4
MAMP的配置:
抗菌肽组合物MAMP:1克MAMP-01+3克MAMP-02+8克MAMP-03。
其他实验方法与实施例1相同。对五种菌的抑菌结果见表4,结果表明抗菌肽组合物MAMP对革兰氏阴性、革兰氏阳性菌和真菌均有抑制活性,并且对于以上五种菌的抑制活性明显好于单一抗菌肽。
表4抗菌肽组合物MAMP的抑菌结果
实施例5
MAMP的配置:
MAMP组合1:1克MAMP-01+5克MAMP-02+10克MAMP-03
MAMP组合2:1克MAMP-01+15克MAMP-02+10克MAMP-03
MAMP组合3:1克MAMP-01+20克MAMP-02+2克MAMP-03
MAMP组合4:1克MAMP-01+20克MAMP-02+30克MAMP-03
其他实验方法与实施例1相同。
对五种菌的抑菌结果见表5,表中列出了MAMP抗菌肽组合1、MAMP抗菌肽组合2、MAMP抗菌肽组合3、MAMP抗菌肽组合4、MAMP抗菌肽组合5的抑菌结果。结果表明五种抗菌肽组合MAMP均对革兰氏阴性、革兰氏阳性菌和真菌均很好的抑制活性,其中MAMP抗菌肽组合5的抑菌活性明显好于其他4种组合。
表5抗菌肽组合物MAMP的抑菌结果
Claims (10)
1.一种抗菌肽组合物,其特征在于:所述抗菌肽组合由三个抗菌肽组成,依次命名为MAMP-01、MAMP-02、MAMP-03,分别为:
(1)MAMP-01为多肽AASDISLL,具有序列表SEQ ID NO:1中氨基酸序列;
该多肽的氨基酸序列具体为:Ala-Ala-Ser-Asp-Ile-Ser-Leu-Leu;
(2)MAMP-02为多肽LDAQSAPLR,具有序列表SEQ ID NO:2中氨基酸序列;该多肽的氨基酸序列具体为:
Leu-Asp-Ala-Gln-Ser-Asp-Pro-Leu-Arg。
(3)MAMP-03为多肽LDTDYKKY,具有序列表SEQ ID NO:3中氨基酸序列;该多肽的氨基酸序列具体为:Leu-Asp-Thr-Asp-Tyr-Lys-Lys-Tyr。
2.按照权利要求1所述的抗菌肽组合物,其特征在于:三个来源于水解牛奶蛋白的抗菌肽组合,命名为MAMP。
3.按照权利要求1或2所述的抗菌肽组合物,三个抗菌肽的比例为MAMP-01:MAMP-02:MAMP-03=1:0-10:5~20(重量比),优选MAMP-01:MAMP-02:MAMP-03=1:0-8:7~15(重量比),更优选MAMP-01:MAMP-02:MAMP-03=1:0-6:8~12(重量比)。
4.一种权利要求1或2或3所述抗菌肽组合物在制备微生物生长抑制剂、抗感染药物、口腔用品或化妆品中的应用。
5.按照权利要求4所述的应用,其特征在于:所述微生物生长抑制剂、抗感染药物、口腔用品或化妆品是以MAMP为活性成份,其中可添加药物学、食品、口腔用品、化妆品中可接受的载体或辅料中的一种或两种以上。
6.按照权利要求4或5所述的应用,其特征在于:所述微生物为革兰氏阴性或革兰氏阳性菌或真菌中的一种或二种。
7.按照权利要求4或5所述的应用,其特征在于:所述感染为由大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变异链球菌、白色念珠菌中的一种或两种以上菌引起感染或菌群失衡。
8.按照权利要求4或5所述的应用,其特征在于:
所述药物为预防和/或降低或治疗由革兰氏阳性或革兰氏阴性菌或真菌中的一种或两种以上引起的口腔疾病的药品;
所述口腔用品为预防和/或降低或治疗由革兰氏阳性或革兰氏阴性菌或真菌中的一种或两种以上引起的口腔疾病的口腔用品。
9.按照权利要求4或5所述的应用,其特征在于:
所述药物为预防和/或降低或治疗由革兰氏阳性或革兰氏阴性菌或真菌中的一种或两种以上引起的皮肤疾病的药品;
所述化妆品为预防和/或降低或治疗由革兰氏阳性或革兰氏阴性菌或真菌中的一种或两种以上引起的皮肤疾病的化妆品。
10.一种权利要求1或2或3所述抗菌肽组合物在制备微生物生长抑制剂、抗感染药物、口腔用品、化妆品的应用,其中可添加药物学、食品、口腔用品、化妆品中可接受的载体或辅料中的一种或两种以上。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110590640.2A CN114149483B (zh) | 2021-05-28 | 2021-05-28 | 一种抗菌肽组合物及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110590640.2A CN114149483B (zh) | 2021-05-28 | 2021-05-28 | 一种抗菌肽组合物及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114149483A true CN114149483A (zh) | 2022-03-08 |
CN114149483B CN114149483B (zh) | 2023-05-30 |
Family
ID=80460945
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110590640.2A Active CN114149483B (zh) | 2021-05-28 | 2021-05-28 | 一种抗菌肽组合物及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114149483B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114149484A (zh) * | 2021-05-28 | 2022-03-08 | 中国科学院大连化学物理研究所 | 一种抗菌肽mamp-03及其应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2145313A1 (en) * | 1992-10-02 | 1994-04-14 | William Shafer | Antimicrobial Peptides |
WO1999031123A1 (en) * | 1997-12-18 | 1999-06-24 | Periodontix, Inc. | Antifungal and antibacterial peptides |
WO2021091369A1 (en) * | 2019-11-07 | 2021-05-14 | N.V. Nutricia | Extensive whey protein hydrolysate with tolerogenic peptides |
CN112824429A (zh) * | 2019-11-21 | 2021-05-21 | 中国科学院大连化学物理研究所 | 一种人乳内源性抗菌多肽及其在在制备抗炎药物中的应用 |
-
2021
- 2021-05-28 CN CN202110590640.2A patent/CN114149483B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2145313A1 (en) * | 1992-10-02 | 1994-04-14 | William Shafer | Antimicrobial Peptides |
WO1999031123A1 (en) * | 1997-12-18 | 1999-06-24 | Periodontix, Inc. | Antifungal and antibacterial peptides |
WO2021091369A1 (en) * | 2019-11-07 | 2021-05-14 | N.V. Nutricia | Extensive whey protein hydrolysate with tolerogenic peptides |
CN112824429A (zh) * | 2019-11-21 | 2021-05-21 | 中国科学院大连化学物理研究所 | 一种人乳内源性抗菌多肽及其在在制备抗炎药物中的应用 |
Non-Patent Citations (4)
Title |
---|
A PIHLANTO-LEPPÄLÄ 等: ""Angiotensin I-converting enzyme inhibitory properties of whey protein digests: concentration and characterization of active peptides "" * |
FISSEHA T WELDERUFAEL 等: ""Production of angiotensin-I-converting enzyme inhibitory peptides from β-lactoglobulin- and casein-derived peptides: an integrative approach"" * |
MARLOES SCHURINK等: ""Novel peptides with tyrosinase inhibitory activity"" * |
XIAOYU WANG 等: ""QSAR study on angiotensin-converting enzyme inhibitor oligopeptides based on a novel set of sequence information descriptors"" * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114149484A (zh) * | 2021-05-28 | 2022-03-08 | 中国科学院大连化学物理研究所 | 一种抗菌肽mamp-03及其应用 |
Also Published As
Publication number | Publication date |
---|---|
CN114149483B (zh) | 2023-05-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Xuan et al. | Antimicrobial peptides for combating drug-resistant bacterial infections | |
CN102871996A (zh) | 一种抗菌药物组合物及其应用 | |
CN112760253B (zh) | 一种植物乳杆菌、抗菌肽及其应用 | |
US20210283215A1 (en) | Bacteriotherapy against proprionibacterium acnes for the treatment of acne | |
CN115317522A (zh) | 一株鼠李糖乳杆菌r7970在制备病原菌抑制产品中的应用 | |
CN114149483B (zh) | 一种抗菌肽组合物及其应用 | |
CN113321708B (zh) | 一种人工设计抗菌肽的制备及其在水产上的应用 | |
CN114456239A (zh) | 生泰素以及由其制备的外用抗菌肽凝胶制剂与应用 | |
CN107236022B (zh) | 细胞穿透肽的亲脂性化合物偶联物及其在抗菌中的应用 | |
CN115850409A (zh) | 一种抗多种病原菌的无前导肽细菌素subticin A3及制备方法与应用 | |
US20060073156A1 (en) | Fosfomycin and n-acetylcysteine for the treatment of biofilms caused by escheric ia coli and other pathogens of the urinary tract | |
CN110711192A (zh) | 色氨酸增强对革兰氏阴性菌杀菌作用的应用 | |
CN117069819B (zh) | 一种黑腹狼蛛抗菌肽lc-amp-i1及其应用 | |
CN112724198A (zh) | 一种抗耐甲氧西林金黄色葡萄球菌抗菌肽及其制备方法和应用 | |
CN110317247B (zh) | 一种抗口腔致病菌多肽及应用 | |
KR102191432B1 (ko) | 유해세균에 대해 항균 활성을 갖는 락토바실러스 타이완엔시스 균주 및 이의 용도 | |
CN116003520B (zh) | 一种副干酪乳杆菌精氨酸生物合成蛋白抗菌肽ngj1d及其应用 | |
CN116687891B (zh) | 苯基氯化硒的应用 | |
WO2023027240A1 (ko) | 물봉선 유래 항균 펩타이드 및 이를 함유하는 항균용 조성물 | |
CN115414342A (zh) | 芬布芬在制备杀灭革兰氏阴性菌药物中的应用 | |
CN118146312A (zh) | 一种抗菌肽kw13及其应用 | |
CN117466974A (zh) | 一种抗微生物肽及其应用 | |
CN113994971A (zh) | 激酶抑制剂在防治农业病原细菌中的用途 | |
CN114149484A (zh) | 一种抗菌肽mamp-03及其应用 | |
CN118452325A (zh) | 一种组合物及其在抑菌和防治奶牛乳房炎中的应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |