CN114763377A - 一种人乳内源性的抗菌肽及其应用 - Google Patents
一种人乳内源性的抗菌肽及其应用 Download PDFInfo
- Publication number
- CN114763377A CN114763377A CN202110047565.5A CN202110047565A CN114763377A CN 114763377 A CN114763377 A CN 114763377A CN 202110047565 A CN202110047565 A CN 202110047565A CN 114763377 A CN114763377 A CN 114763377A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- tldggfiyeaglapyklrpv
- leu
- gly
- peptide
- 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
- 235000020256 human milk Nutrition 0.000 title claims abstract description 15
- 210000004251 human milk Anatomy 0.000 title claims abstract description 15
- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims description 14
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 42
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 31
- 229920001184 polypeptide Polymers 0.000 claims abstract description 25
- 239000003814 drug Substances 0.000 claims abstract description 11
- 244000005700 microbiome Species 0.000 claims abstract description 7
- 206010022678 Intestinal infections Diseases 0.000 claims abstract description 5
- 235000013305 food Nutrition 0.000 claims abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 4
- 235000013350 formula milk Nutrition 0.000 claims abstract description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 3
- 230000000813 microbial effect Effects 0.000 claims abstract 3
- 241000191967 Staphylococcus aureus Species 0.000 claims description 15
- 241000588724 Escherichia coli Species 0.000 claims description 14
- 229940079593 drug Drugs 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000002924 anti-infective effect Effects 0.000 claims description 3
- 208000015181 infectious disease Diseases 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 102000044503 Antimicrobial Peptides Human genes 0.000 claims 2
- 108700042778 Antimicrobial Peptides Proteins 0.000 claims 2
- 239000003966 growth inhibitor Substances 0.000 claims 2
- 239000000463 material Substances 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 abstract description 8
- 241000894006 Bacteria Species 0.000 abstract description 6
- 150000001875 compounds Chemical class 0.000 abstract description 4
- 241000192125 Firmicutes Species 0.000 abstract description 2
- 239000000654 additive Substances 0.000 abstract description 2
- 230000000996 additive effect Effects 0.000 abstract description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 13
- 150000001413 amino acids Chemical class 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 8
- 230000003385 bacteriostatic effect Effects 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 102000007079 Peptide Fragments Human genes 0.000 description 5
- 108010033276 Peptide Fragments Proteins 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 235000019253 formic acid Nutrition 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 3
- XXMFJKNOJSDQBM-UHFFFAOYSA-N 2,2,2-trifluoroacetic acid;hydrate Chemical compound [OH3+].[O-]C(=O)C(F)(F)F XXMFJKNOJSDQBM-UHFFFAOYSA-N 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 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
- 238000004458 analytical method Methods 0.000 description 2
- 229940124350 antibacterial drug Drugs 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 230000007717 exclusion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 102000011632 Caseins Human genes 0.000 description 1
- 108010076119 Caseins Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241001646716 Escherichia coli K-12 Species 0.000 description 1
- 101000798114 Homo sapiens Lactotransferrin Proteins 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 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
- 239000002671 adjuvant Substances 0.000 description 1
- -1 aliphatic amino acid Chemical class 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000013480 data collection Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000001516 effect on protein Effects 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 102000050459 human LTF Human genes 0.000 description 1
- 210000004293 human mammary gland Anatomy 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000005596 ionic collisions Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 235000021247 β-casein Nutrition 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/79—Transferrins, e.g. lactoferrins, ovotransferrins
-
- 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
- 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
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Communicable Diseases (AREA)
- Pharmacology & Pharmacy (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明涉及一种来自人乳内源性的具有微生物抑制活性的多肽化合物TLDGGFIYEAGLAPYKLRPV，其氨基酸序列为Thr‑Leu‑Asp‑Gly‑Gly‑Phe‑Ile‑Tyr‑Glu‑Ala‑Gly‑Leu‑Ala‑Pro‑Tyr‑Lys‑Leu‑Arg‑Pro‑Val。多肽TLDGGFIYEAGLAPYKLRPV具有对革兰氏阳性菌和革兰氏阴性菌的广泛抑制活性，作为预防和/或降低新生儿由微生物引起的肠道感染疾病的药物和/或保健品或者新型婴儿配方食品的添加剂具有良好的应用前景。
Description
技术领域
本发明涉及一种来源于人乳的内源性抗菌多肽TLDGGFIYEAGLAPYKLRPV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。
母乳作为婴儿最佳的天然食物，是丰富的蛋白质和多肽资源，兼顾抗菌肽的优势及人乳优质安全的蛋白来源两个特质。人乳腺中的内源性酶对蛋白质有较强的水解作用，而来源于人乳中稳定存在的内源性抗菌肽，其本身的存在就解决了抗菌肽进入人体后易被进一步水解的问题，作为抗菌药物具有良好的潜质。因此，从人乳内源性多肽出发进一步寻找具有抑菌活性的肽会为临床上对婴幼儿抗感染的治疗提供新的治疗手段。
发明内容
本发明的目的是提供一种内源性抗菌多肽TLDGGFIYEAGLAPYKLRPV，在制备预防和/或降低由大肠杆菌和金黄色葡萄球菌引起的肠道感染疾病的药物和/或保健品或者新型婴儿配方食品中的应用。
为实现上述目的，本发明以所述多肽TLDGGFIYEAGLAPYKLRPV为抑制微生物生长活性的有效成份。
其具有序列表SEQ ID NO：1中氨基酸序列；多肽TLDGGFIYEAGLAPYKLRPV为抗大肠杆菌和金黄色葡萄球菌药物的活性成份，其中可添加药物学上可接受的载体或辅料。
具有对大肠杆菌和金黄色葡萄球菌抑制活性的多肽化合物TLDGGFIYEAGLAPYKLRPV的氨基酸序列为Thr-Leu-Asp-Gly-Gly-Phe-Ile-Tyr-Glu-Ala-Gly-Leu-Ala-Pro-Tyr-Lys-Leu-Arg-Pro-Val。分子量2180.49Da，白色粉末状，易溶于水，对大肠杆菌和金黄色葡萄球菌的生长具有很强的抑制作用。
本发明与现有技术相比，具有如下有益效果：
本发明从人乳中获得并确定了上述多肽的结构，首次验证了上述多肽具有较好的抑菌活性，因此作为预防和/或降低新生儿由微生物引起的肠道感染疾病的药物和/或保健品或者新型婴儿配方食品的添加剂具有良好的应用前景。
附图说明
图1本发明多肽对大肠杆菌和金黄葡萄球菌的抑菌结果。
图1本发明多肽对大肠杆菌和金黄葡萄球菌的抑菌结果图，其左面为A，多肽TLDGGFIYEAGLAPYKLRPV抑制大肠杆菌的实验结果图；右面为B，多肽TLDGGFIYEAGLAPYKLRPV抑制金黄色葡萄球菌的实验结果图。
具体实施方式
实施例1
人乳内源肽的制备及鉴定。
(1)样品制备
取200μL人乳原液以水稀释至人乳原液体积的10倍，水浴95℃条件下5min变性，以截留分子量10K Da的超滤管在14000×g、20℃条件下超滤20min，超滤液为内源肽样品。
(2)LC-MS/MS分析
内源肽样品按如下步骤以C18柱(Waters Oasis HLB SPE柱)除盐：C18柱用1.5mL甲醇活化后加入1.5mL 0.1％(V/V)TFA(三氟乙酸)-H2O溶液平衡，将样品以0.1％(V/V)TFA-H2O溶液复溶后加入C18柱中，以1.5mL 80％(V/V)ACN(乙腈)/0.1％(V/V)TFA-H2O溶液进行洗脱，收集洗脱液分装冻干于-80℃保存。脱盐后的样品以0.1％(V/V)FA(甲酸)-H2O溶解并制备成浓度为0.2g/L的溶液，上样量为8μL，加载到15cm毛细管分析柱(内径为180μm)进行质谱分析。LC-MS/MS系统由Agilent 1100HPLC系统和LTQ-Orbitrap Velos质谱仪组成。流动相A为含0.1％甲酸的水溶液，流动相B为含0.1％甲酸(V/V)和98％乙腈(V/V)的水溶液。梯度洗脱程序如下：0～80min，5～25％B；80～95min，25％～35％B；95～97min，35％～90％B；97～107min，90％B；107～109min，90％～0％B；109～126min，0％B；流速70μL/min。在Orbitrap质量分析仪中以60000的分辨率获得了完整的质谱扫描图(m/z 400～2000)。对其中最强的15个离子进行离子碰撞解离(CID)，再进行MS/MS扫描，扫描范围在m/z400～2000。动态排除功能设置如下：重复2；持续时间30s；排除持续时间为60s。系统控制和数据收集由Xcalibur软件进行。每个样品进行三次质谱分析。
(3)数据检索
Xcalibur采集的*.RAW文件用Thermo Proteome Discoverer(v1.4)转换成*.MGF格式，之后用Mascot 2.5.1软件在人(Human spains，蛋白质数目为22427)蛋白数据库(http://www.uniprot.org/)中进行检索。搜库参数如下：不设置酶切、最大漏切数和固定修饰，可变修饰设置为甲硫氨酸的氧化(+15.9949Da)。母离子的质量容忍偏差为20ppm，碎片离子为0.8Da。导出肽段时控制假阳性率(FDR)<1％，对导出结果Score>20的肽段视为有效数据进行分析。
(4)LC-MS/MS得到目标肽段信息
在人乳内源肽样品中鉴定到一千条以上的肽段信息，其中约50％以上来源于β-酪蛋白。来源于人乳内源性的多肽TLDGGFIYEAGLAPYKLRPV蛋白质前体为人乳铁蛋白(f77-96)，肽段等电点为5.73，分子质量为2180.49Da，具有20个氨基酸，较短的序列及较低的相对分子质量，结合质谱鉴定结果表明该序列可以稳定存在不易继续被蛋白酶酶解。通过在线工具获取SEQ ID NO：1所示氨基酸序列的生物学信息表示，该肽段的不稳定系数为46.18，脂肪族氨基酸指数和亲水性分别为102.50和0.170，表现为疏水性。
SEQ ID No.1的信息
(a)序列特征
*长度：20氨基酸
*类型：氨基酸
*链型：单链
(b)分子类型：蛋白
序列描述：
SEQ ID No.1
TLDGGFIYEAGLAPYKLRPV
实施例2多肽的抗菌活性
委托上海杰肽生物科技有限公司通过固相法合成多肽TLDGGFIYEAGLAPYKLRPV，纯度为98.20％。以革兰氏阴性菌大肠杆菌、革兰氏阳性菌金色葡萄球菌为靶标菌种，考察多肽TLDGGFIYEAGLAPYKLRPV的抗菌活性。
(1)菌株及复苏
大肠杆菌(Escherichia coli K12)和金黄色葡萄球菌(Staphylococcus aureus)均以甘油保藏法保存于-80℃，实验前将两种菌分别接种到液体LB培养基和TSB培养基中，于37℃条件下培养12h使菌种复苏，重复上述复苏操作2-3次，使菌株恢复活力。
(2)培养基
抗菌实验使用直径为90mm的圆形培养皿。双层培养基，下层以15mL2％(m/V，g/mL)的琼脂水溶液做支撑，上层分别加入5mL含0.7％(m/V，g/mL)琼脂、菌浓度为1×10^5CFU/mL的LB培养基和TSB培养基，将制备好的两种培养基密封后于4℃保存。
(3)抗菌实验
在平板上层制作出直径2μm的孔，各孔间距约3.5cm。合成的标肽分装成5mg/支，每支加入50μL无菌水，完全溶解后向孔中加入标肽水溶液35μL，每孔实际加入标肽量3.5mg。以水作空白对照，加样体积与标肽样品同为35μL，样品设置两个平行孔。加样后的平板于4℃冰箱中静置3h，待孔中样液完全吸收扩散后取出，在37℃恒温培养箱中倒置培养12h，观察是否有抑菌圈出现，测量抑菌圈大小并记录。
(4)实验结果
对大肠杆菌和金黄葡萄球菌的抑菌结果见表1和图1，表中列出了目标肽段和空白对照的抑菌圈直径，两个平行孔抑菌结果分别记作“1”和“2”。结果表明多肽TLDGGFIYEAGLAPYKLRPV对革兰氏阴性菌和革兰氏阳性菌均有生长抑制活性，对金黄色葡萄球菌的生长抑制效果要优于大肠杆菌。
表1多肽TLDGGFIYEAGLAPYKLRPV的抑菌结果
“-”表示没有抑菌活性。
Claims (6)
1.人乳内源性的抗菌肽，其特征在于：所述抗菌肽为多肽TLDGGFIYEAGLAPYKLRPV，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，Thr-Leu-Asp-Gly-Gly-Phe-Ile-Tyr-Glu-Ala-Gly-Leu-Ala-Pro-Tyr-Lys-Leu-Arg-Pro-Val。
2.一种权利要求1所述抗菌肽在制备微生物生长抑制剂或抗感染药物中的应用。
3.按照权利要求2所述的应用，其特征在于：所述微生物生长抑制剂或抗感染药物是以多肽TLDGGFIYEAGLAPYKLRPV为活性成份，其中可添加药物学上可接受的载体或辅料。
4.按照权利要求2所述的应用，其特征在于：所述抗菌肽在制备预防和/或降低由大肠杆菌和/或金黄色葡萄球菌引起的肠道感染疾病的药物和/或保健品或者婴儿配方食品中的应用。
5.按照权利要求2所述的应用，其特征在于：所述微生物为大肠杆菌和金黄色葡萄球菌中的一种或二种以上。
6.按照权利要求2所述的应用，其特征在于：所述感染为由大肠杆菌和/或金黄色葡萄球菌引起的肠道感染。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110047565.5A CN114763377B (zh) | 2021-01-14 | 2021-01-14 | 一种人乳内源性的抗菌肽及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110047565.5A CN114763377B (zh) | 2021-01-14 | 2021-01-14 | 一种人乳内源性的抗菌肽及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114763377A true CN114763377A (zh) | 2022-07-19 |
| CN114763377B CN114763377B (zh) | 2023-11-03 |
Family
ID=82363318
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110047565.5A Active CN114763377B (zh) | 2021-01-14 | 2021-01-14 | 一种人乳内源性的抗菌肽及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114763377B (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111072770A (zh) * | 2019-12-20 | 2020-04-28 | 华中农业大学 | 卵转铁蛋白抗菌肽及其制备方法 |
| CN111848739A (zh) * | 2020-06-30 | 2020-10-30 | 河南科技学院 | 一种抗菌肽lj-2及其应用 |
-
2021
- 2021-01-14 CN CN202110047565.5A patent/CN114763377B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111072770A (zh) * | 2019-12-20 | 2020-04-28 | 华中农业大学 | 卵转铁蛋白抗菌肽及其制备方法 |
| CN111848739A (zh) * | 2020-06-30 | 2020-10-30 | 河南科技学院 | 一种抗菌肽lj-2及其应用 |
Non-Patent Citations (1)
| Title |
|---|
| 于文皓: "人乳内源肽组成及其与泌乳期的相关性研究", 中国优秀硕士论文全文数据库（电子期刊）医药卫生科技辑, no. 2020, pages 18 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114763377B (zh) | 2023-11-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112824429B (zh) | 一种人乳内源性抗菌多肽及其在制备抗炎药物中的应用 | |
| CN107298706B (zh) | 一种荷斯坦奶牛脾脏源抗菌肽及其制备方法与应用 | |
| CN109553657B (zh) | 一种非完美两亲性肽w4及其制备方法和应用 | |
| CN112824430B (zh) | 一种人乳内源性的抗菌肽及其应用 | |
| CN114763377A (zh) | 一种人乳内源性的抗菌肽及其应用 | |
| CN105801669A (zh) | 一种杂合式抗生物型抗菌肽及制备方法和应用 | |
| CN112961210B (zh) | 一种人乳内源性抗菌多肽及其制备抗炎药物中的应用 | |
| CN112851762B (zh) | 一种人乳内源性抗菌多肽及其在制备抗炎药物中的应用 | |
| CN112851799B (zh) | 人乳内源性抗菌多肽及其在制备抗炎药物中的应用 | |
| CN112851798B (zh) | 一种人乳内源性抗菌多肽和其在制备抗炎药物中的应用 | |
| CN112851752B (zh) | 人乳内源性抗菌多肽和其在制备抗炎药物中的应用 | |
| CN118063544A (zh) | 一种抗菌肽或组合物及其应用 | |
| CN114149486B (zh) | 一种抗菌肽mamp-01及其应用 | |
| CN114149484B (zh) | 一种抗菌肽mamp-03及其应用 | |
| CN114149485B (zh) | 一种抗菌肽mamp-02及其应用 | |
| CN116375877B (zh) | 一种细胞穿透抗菌肽pw2及其制备方法和应用 | |
| CN113956340B (zh) | 一种衍生自羊源抗菌肽rlr及其制备方法和应用 | |
| CN117886913A (zh) | 乳源抗菌多肽及其药物和应用 | |
| CN114149483B (zh) | 一种抗菌肽组合物及其应用 | |
| CN108524945B (zh) | 一种庆大霉素改性壳聚糖的制备方法 | |
| 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 | |
| CN116693607A (zh) | 抗菌多肽及应用和细菌生长抑制剂、抗感染药物或保健品 | |
| CN109758572A (zh) | N6衍生肽的应用 | |
| CN116693619A (zh) | 抗菌多肽与应用以及抑菌剂、抗感染药物或保健品 | |
| CN116694599A (zh) | 抗菌多肽和应用及抑菌剂、抗感染药物或保健品 |
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 |