CN103772494A - 一种丁酸梭菌分泌的抗菌肽及其制备方法和应用 - Google Patents
一种丁酸梭菌分泌的抗菌肽及其制备方法和应用 Download PDFInfo
- Publication number
- CN103772494A CN103772494A CN201410030648.3A CN201410030648A CN103772494A CN 103772494 A CN103772494 A CN 103772494A CN 201410030648 A CN201410030648 A CN 201410030648A CN 103772494 A CN103772494 A CN 103772494A
- Authority
- CN
- China
- Prior art keywords
- antibacterial peptide
- preparation
- antibacterial
- exchange
- clostridium butylicum
- 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 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 241000193171 Clostridium butyricum Species 0.000 title abstract description 4
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 12
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 4
- 238000012870 ammonium sulfate precipitation Methods 0.000 claims abstract description 3
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 3
- 239000002808 molecular sieve Substances 0.000 claims abstract description 3
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 claims abstract description 3
- 241000193403 Clostridium Species 0.000 claims description 24
- 230000028327 secretion Effects 0.000 claims description 17
- 230000000845 anti-microbial effect Effects 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 238000005342 ion exchange Methods 0.000 claims description 6
- 239000007853 buffer solution Substances 0.000 claims description 5
- 229920001184 polypeptide Polymers 0.000 claims description 5
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 5
- 230000003115 biocidal effect Effects 0.000 claims description 4
- 238000000502 dialysis Methods 0.000 claims description 4
- 230000002949 hemolytic effect Effects 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 3
- 230000003248 secreting effect Effects 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 241000588724 Escherichia coli Species 0.000 claims description 2
- 239000013543 active substance Substances 0.000 claims description 2
- 238000003453 ammonium sulfate precipitation method Methods 0.000 claims description 2
- 125000002091 cationic group Chemical group 0.000 claims description 2
- 239000012228 culture supernatant Substances 0.000 claims description 2
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims description 2
- 206010018910 Haemolysis Diseases 0.000 abstract description 4
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 4
- 230000008588 hemolysis Effects 0.000 abstract description 4
- 238000005341 cation exchange Methods 0.000 abstract description 2
- 239000003814 drug Substances 0.000 abstract description 2
- 241000282887 Suidae Species 0.000 abstract 1
- 230000001205 effect on erythrocytes Effects 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
- 241000282898 Sus scrofa Species 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 230000009182 swimming Effects 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 239000000872 buffer Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000005277 cation exchange chromatography Methods 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000006641 stabilisation Effects 0.000 description 3
- 238000011105 stabilization Methods 0.000 description 3
- 206010010356 Congenital anomaly Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 238000009631 Broth culture Methods 0.000 description 1
- 241001360526 Escherichia coli ATCC 25922 Species 0.000 description 1
- 241001646716 Escherichia coli K-12 Species 0.000 description 1
- FCKPEGOCSVZPNC-WHOFXGATSA-N Gly-Ile-Phe Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 FCKPEGOCSVZPNC-WHOFXGATSA-N 0.000 description 1
- WZPIKDWQVRTATP-SYWGBEHUSA-N Ile-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)[C@@H](C)CC)C(O)=O)=CNC2=C1 WZPIKDWQVRTATP-SYWGBEHUSA-N 0.000 description 1
- VGPCJSXPPOQPBK-YUMQZZPRSA-N Leu-Gly-Ser Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O VGPCJSXPPOQPBK-YUMQZZPRSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 1
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 1
- 241000751182 Staphylococcus epidermidis ATCC 12228 Species 0.000 description 1
- HPQHHRLWSAMMKG-KATARQTJSA-N Thr-Lys-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)O)N)O HPQHHRLWSAMMKG-KATARQTJSA-N 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- LGEPIBQBGZTBHL-SXNHZJKMSA-N Trp-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N LGEPIBQBGZTBHL-SXNHZJKMSA-N 0.000 description 1
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 1
- 108010078114 alanyl-tryptophyl-alanine Proteins 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 238000005142 microbroth dilution method Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000009781 safety test method Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012360 testing method Methods 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
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/33—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Clostridium (G)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/195—Antibiotics
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Polymers & Plastics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种丁酸梭菌分泌的抗菌肽及其氨基酸序列和应用。所述的抗菌肽氨基酸序列为序列表SEQIDNO:1所示，分子量为2264.6道尔顿，等电点为8.64。其制备方法是：硫酸铵沉淀培养基中丁酸梭菌分泌的抗菌肽，利用阳离子交换、分子筛层析和反向高效液相色谱法纯化抗菌肽。该抗菌肽具有显著的抗菌活性，且不会对猪的红细胞产生溶血作用，可用于开发制备饲用抗菌药物。
Description
技术领域
本发明属于生物技术，具体的，本发明涉及一种丁酸梭菌分泌的抗菌肽及其制备方法和应用。
背景技术
抗菌肽是机体先天免疫的重要组成成分，是许多生物体抵抗外来致病菌侵袭的第一道屏障。由于其独特且多样的生物学功能和作用机制引起了科学界的广泛关注和研究，一方面研究抗菌肽在先天免疫中发挥作用的生物学基础，一方面将其作为设计新型抗菌、抗感染制剂的分子模板。抗菌肽是自然进化的产物，和微生物一起已经存在了亿万年，尽管经过长期的共同进化，但抗菌肽并没有丢失其杀灭或抑制微生物的能力，或产生使微生物逃逸抗菌肽的能力，所以说抗菌肽是研发新型高效抗生素替代品的重要突破。
丁酸梭菌是一种益生菌，具有维持肠道微生态平衡、提高免疫力、抗癌、产生营养物质等功能。丁酸梭菌开发利用前景广阔，目前已应用于食品、医药、农牧、化工等诸多领域。但由于丁酸梭菌培养条件严格，需要在严格厌氧条件下才能生长，且分泌的成分复杂，难被分离纯化，因此目前尚无从丁酸梭菌中分离纯化抗菌肽的报道。
发明内容
本发明的目的在于提供一种新的具有广谱抗菌的丁酸梭菌分泌的抗菌肽及其制备方法和在饲料中的应用。
为了实现本发明，本发明提供如下技术方案：
一种丁酸梭菌分泌的抗菌肽，从丁酸梭菌分泌产物中分离达到的一种单链多肽，分子量是2264.6，等电点8.64，多肽的氨基酸序列如SEQ ID NO:1所示为：PSAWQITKCAGSIAWALGSGIF，该抗菌肽具有抗菌活性，且不会对猪的红细胞产生溶血作用。
所述的抗菌肽的制备方法，将丁酸梭菌置于严格厌氧条件培养24小时后离心，收集培养上清并用硫酸铵沉淀法富集所述的抗菌肽，经离子交换、分子筛层析和反相高效液相色谱纯化。
所述的离子交换为：将硫酸铵沉淀后得到的样品透析，上样于阳离子交换柱，用含0~0.5M NaCl的缓冲液梯度洗脱，收集洗脱的蛋白。
所述的分子筛层析为：将离子交换得到的活性成分透析，上样于分子筛层析柱，用缓冲液洗脱，收集第四个峰。
所述的反向高效液相色谱为：将分子筛层析得到的活性物质上样于C18柱，用含0.1%的乙腈从10~60%梯度洗脱，收集45min处峰。
所述的抗菌肽的应用，作为饲料的抗菌制剂。
本发明采用飞行质谱法测定分子量，使用ExPASy在线工具估算等电点，自动氨基酸测序仪测定氨基酸的序列结构。
下面用本发明丁酸梭菌分泌的抗菌肽的抗菌试验和安全性试验说明本发明的有益效果：地衣芽孢杆菌分泌的抗菌肽对革兰氏阳性菌和阴性菌均有较高的抗菌活性，对E. coli K 88的最小抑菌浓度为32μg/mL，对S. aureus ATCC25923的最小抑菌浓度为64 μg/mL；安全性高，抗菌肽浓度高达256 μg/mL对猪红细胞也无溶血作用。
附图说明
图1 阳离子交换色谱纯化丁酸梭菌分泌的抗菌肽SDS-PAGE图，（泳道1，蛋白marker；泳道2，丁酸梭菌分泌的总蛋白；泳道3，阳离子交换色谱纯化后得到的蛋白）；
图2 分子筛层析色谱纯化丁酸梭菌分泌的抗菌肽SDS-PAGE图，（泳道1，蛋白marker；泳道2，阳离子交换色谱纯化后得到的蛋白；泳道3，分子筛层析色谱纯化后得到的蛋白）；
图3丁酸梭菌分泌的抗菌肽对猪红细胞溶血率测定。
具体实施方式
以下结合附图和实施例对本发明做进一步的说明。
实施例1 阳离子交换纯化丁酸梭菌分泌的抗菌肽
1) 装柱
2) 10体积的低盐缓冲液，至紫外吸收值稳定
3) 将样品注射至进样柱内，调整进样流速为1 ml/min；
4) 10体积的低盐缓冲液，至紫外吸收值稳定
5) 0~0.5M NaCl氯化钠梯度洗脱
6) 收集样品，用Bradford试剂盒测定蛋白浓度，并用低盐缓冲液将蛋白浓度调整为1mg/ml，取5 μg样品用于NuPAGE分析，如图1所示，其余保存于﹣80℃超低温冰箱。
实施例2 分子筛层析纯化丁酸梭菌分泌的抗菌肽
1) 装柱
2) 10体积的缓冲液平衡，至紫外吸收值稳定
3) 将样品注射至进样柱内，调整进样流速为0.1 ml/min；
4）缓冲液洗脱，收集第四个峰
5) 收集样品，用Bradford试剂盒测定蛋白浓度。用琼脂糖孔穴扩散法检测每个峰的抗菌活性，用低盐缓冲液将蛋白浓度调整为1mg/ml，取5 μg样品用于NuPAGE分析，如图2所示，其余保存于﹣80℃超低温冰箱。
实施例3 丁酸梭菌分泌的抗菌肽对猪红细胞溶血率测定
1）将抗菌肽在96孔板上进行系列稀释，终浓度为256、128、64、32、16、8、4 μg/mL，每个浓度10uL，共七个浓度，同时设置三个重复；
2）90uL/孔的红细胞悬液加入到稀释好的多肽孔中，设置阳性对照孔为1%红细胞悬液中加入10uL 10%的Triton X-100（终浓度为1%）；阴性对照孔位1%红细胞悬液中加入10uL的1×PBS；
3）将96孔板置于37℃，5% CO2环境中培养18-24h；
4）培养结束后，将96孔板在离心机无制动停机状态下1000-1500rpm离心20min；
小心吸取上清至一新平底透明96孔培养板中，用酶标仪测定各孔在414nm和546nm下的吸光度；软件分析得到OD414nm、OD546nm和Delta OD值；通过下列公式计算抗菌肽对猪红细胞的溶血率，结果如图3所示：丁酸梭菌分泌的抗菌肽对猪红细胞无溶血作用。
实施例4 丁酸梭菌分泌的抗菌肽抗菌活性的测定
抗菌活性的测定采用最小抑菌浓度（MIC）法，方法如下：先向无菌96孔圆底板中加入90μL制备好的菌悬液，再逐一加入10μL相应浓度的肽稀释液或抗生素稀释液，待测肽的终浓度分别为256、128、64、32、16、8、4、2、1、0.5、0.25 μg/mL；待测抗生素的终浓度分别为64、32、16、8、4、2、1、0.5、0.25 、0.125、0.0625 μg/mL；另外设阳性对照孔为不加抗菌物质，只加100μL菌悬液，阴性对照孔加100μL MH肉汤培养基；将96孔板置于37℃生化培养箱中保湿静置培养18-24小时；肠道益生菌培养板置于厌氧培养箱中厌氧培养24-48h；每种抗菌物质做三次重复；培养完后观察各孔底部是否有细菌沉淀产生，无肉眼可见细菌沉淀的最小浓度可判定为抗菌物质的MIC。结果如下表1所示：
表1微量肉汤稀释法测定重组抗菌肽CBF的最小抑菌浓度
| 菌种 | 最小抑菌浓度 (μg/ml) |
| 革兰氏阴性菌 | |
| E. coli ATCC25922 | 32 |
| E. coli K 88 | 32 |
| E. coli K 12 | 64 |
| P. aeruginosa CMCC27853 | 64 |
| 革兰氏阳性菌 | |
| S. aureus ATCC25923 | 64 |
| S. epidermidis ATCC12228 | 64 |
SEQUENCE LISTING
<110> 浙江大学
<120> 一种丁酸梭菌分泌的抗菌肽及其制备方法和应用
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> PRT
<213> Clostridium butyricum
<400> 1
Pro Ser Ala Trp Gln Ile Thr Lys Cys Ala Gly Ser Ile Ala Trp Ala
1 5 10 15
Leu Gly Ser Gly Ile Phe
20
Claims (6)
1. 一种丁酸梭菌分泌的抗菌肽，其特征是，从丁酸梭菌分泌产物中分离达到的一种单链多肽，分子量是2264.6，等电点8.64，多肽的氨基酸序列如SEQ ID NO:1所示为：PSAWQITKCAGSIAWALGSGIF，该抗菌肽具有抗菌活性，对E. coli K 88的最小抑菌浓度为32 μg/mL，S. aureus最小抑菌浓度为64 μg/ml，且对猪的红细胞不会产生溶血作用。
2. 一种根据权利要求1所述的抗菌肽的制备方法，其特征是，将丁酸梭菌置于严格厌氧条件培养24小时后离心，收集培养上清并用硫酸铵沉淀法富集所述的抗菌肽，经离子交换、分子筛层析和反相高效液相色谱纯化。
3.根据根据权利要求2所述的制备方法，其特征是，所述的离子交换为：将硫酸铵沉淀后得到的样品透析，上样于阳离子交换柱，用含0~0.5M NaCl的缓冲液梯度洗脱，收集洗脱的蛋白。
4.根据根据权利要求2所述的制备方法，其特征是，所述的分子筛层析为：将离子交换得到的活性成分透析，上样于分子筛层析柱，用缓冲液洗脱，收集第四个峰。
5.根据根据权利要求2所述的制备方法，其特征是，所述的反向高效液相色谱为：将分子筛层析得到的活性物质上样于C18柱，用含0.1%的乙腈从10~60%梯度洗脱，收集45min处峰。
6. 一种根据权利要求1所述的抗菌肽的应用，作为饲料的抗菌制剂。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410030648.3A CN103772494B (zh) | 2014-01-23 | 2014-01-23 | 一种丁酸梭菌分泌的抗菌肽及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410030648.3A CN103772494B (zh) | 2014-01-23 | 2014-01-23 | 一种丁酸梭菌分泌的抗菌肽及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103772494A true CN103772494A (zh) | 2014-05-07 |
| CN103772494B CN103772494B (zh) | 2015-04-15 |
Family
ID=50565307
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410030648.3A Active CN103772494B (zh) | 2014-01-23 | 2014-01-23 | 一种丁酸梭菌分泌的抗菌肽及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103772494B (zh) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106721287A (zh) * | 2016-12-12 | 2017-05-31 | 天津市畜牧兽医研究所 | 一种用于调节仔猪肠道屏障功能的配合饲料及应用 |
| CN109627314A (zh) * | 2019-01-02 | 2019-04-16 | 浙江亮肽生物技术有限公司 | 具有选择性抗大肠杆菌活性的抗菌肽及其活性优化修饰物 |
| 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 |
| CN111437877A (zh) * | 2020-04-22 | 2020-07-24 | 陕西延长石油（集团）有限责任公司 | 一种Cu/Zr双金属骨架型高硅β分子筛催化剂及其制备方法与应用 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008199929A (ja) * | 2007-02-19 | 2008-09-04 | Miyarisan Pharmaceutical Co Ltd | クロストリジウム・ブチリカムのバクテリオシンをコードする遺伝子およびそのタンパク質 |
-
2014
- 2014-01-23 CN CN201410030648.3A patent/CN103772494B/zh active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008199929A (ja) * | 2007-02-19 | 2008-09-04 | Miyarisan Pharmaceutical Co Ltd | クロストリジウム・ブチリカムのバクテリオシンをコードする遺伝子およびそのタンパク質 |
Non-Patent Citations (1)
| Title |
|---|
| 孙雪文: "ZK-I菌发酵液中抗真菌活性化合物的纯化与部分特性研究", 《微生物学通报》, 31 December 2007 (2007-12-31), pages 88 - 91 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106721287A (zh) * | 2016-12-12 | 2017-05-31 | 天津市畜牧兽医研究所 | 一种用于调节仔猪肠道屏障功能的配合饲料及应用 |
| CN109627314A (zh) * | 2019-01-02 | 2019-04-16 | 浙江亮肽生物技术有限公司 | 具有选择性抗大肠杆菌活性的抗菌肽及其活性优化修饰物 |
| 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 |
| CN111437877A (zh) * | 2020-04-22 | 2020-07-24 | 陕西延长石油（集团）有限责任公司 | 一种Cu/Zr双金属骨架型高硅β分子筛催化剂及其制备方法与应用 |
| CN111437877B (zh) * | 2020-04-22 | 2023-03-10 | 陕西延长石油（集团）有限责任公司 | 一种Cu/Zr双金属骨架型高硅β分子筛催化剂及其制备方法与应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103772494B (zh) | 2015-04-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103772494B (zh) | 一种丁酸梭菌分泌的抗菌肽及其制备方法和应用 | |
| CN105175518B (zh) | 凝结芽孢杆菌fm603产生的细菌素及其制备方法 | |
| CN105176874B (zh) | 凝结芽孢杆菌fm603及其应用 | |
| CN103772490B (zh) | 一种地衣芽孢杆菌分泌的抗菌肽及其制备方法和应用 | |
| de Melo et al. | HGA-2, a novel galactoside-binding lectin from the sea cucumber Holothuria grisea binds to bacterial cells | |
| CN107857803A (zh) | 天然抗菌肽及其应用 | |
| Pérez-Giménez et al. | Soybean lectin enhances biofilm formation by Bradyrhizobium japonicum in the absence of plants | |
| CN104592360A (zh) | 碱性抗菌肽及其靶向设计和应用 | |
| CN115536737B (zh) | 一种眼镜王蛇抗菌肽oh-cath30在制备细菌生长抑制剂中的应用 | |
| CN103122347B (zh) | 一种杀灭葡萄球菌的裂解酶及其应用 | |
| BRPI0619512A2 (pt) | peptìdeos indutores de bacteriocina, bem como processo para estimular produção aumentada de bacteriocina | |
| Payne et al. | Selective effects of histidine‐rich polypeptides on the aggregation and viability of Streptococcus mutans and Streptococcus sanguis | |
| Zheng et al. | Novel family of antimicrobial peptides from the skin of Rana shuchinae | |
| CN102382186B (zh) | 衍生自线性鸡β-防御素4(RL38)的抗菌肽GLI23及其制备方法 | |
| CN101328207A (zh) | 一种桔小实蝇抗菌肽及其制备方法和应用 | |
| CN106749588A (zh) | 一种具有杀菌活性的仿刺参肽聚糖识别蛋白及其制备方法和应用 | |
| CN102276729A (zh) | 抗菌肽lf-th及其大肠杆菌重组制备方法 | |
| CN104211804A (zh) | 一种抗金黄色葡萄球菌多克隆抗体的制备与应用 | |
| Odekina et al. | Antimicrobial and antioxidant activities of novel marine bacteria (Bacillus 2011SOCCUF3) isolated from marine sponge (Spongia officinalis) | |
| CN103131686A (zh) | 一种二型肽聚糖识别蛋白及其制备和应用 | |
| Muslim | Improving of antibacterial activity for antibiotics by purified and characterized lectin from Acinetobacter baumannii | |
| Rafiq et al. | Purification and partial characterization of a fructose-binding lectin from the leaves of Euphorbia helioscopia | |
| CN110317253B (zh) | 一种鸡源抗菌肽及其制备方法 | |
| CN112500475B (zh) | 类人乳铁蛋白肽及其应用 | |
| CN107298707A (zh) | 一种类Bac5抗菌肽及其应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent of invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Wang Yizhen Inventor after: Luan Chao Inventor after: Wang Tenghao Inventor after: Feng Jie Inventor after: Du Huahua Inventor after: Wang Xinxia Inventor after: Lu Zeqing Inventor after: Yang Caimei Inventor before: Wang Yizhen Inventor before: Luan Chao |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: WANG YIZHEN LUAN CHAO TO: WANG YIZHEN LUAN CHAO WANG TENGHAO FENG JIE DU HUAHUA WANG XINXIA LU ZEQING YANG CAIMEI |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |