CN106818746B - N15多肽在制备蓝藻抑制剂中的用途 - Google Patents
N15多肽在制备蓝藻抑制剂中的用途 Download PDFInfo
- Publication number
- CN106818746B CN106818746B CN201611169815.8A CN201611169815A CN106818746B CN 106818746 B CN106818746 B CN 106818746B CN 201611169815 A CN201611169815 A CN 201611169815A CN 106818746 B CN106818746 B CN 106818746B
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- cyanobacteria
- purposes
- blue algae
- heterocyst
- 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.)
- Active
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 25
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 21
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 21
- 241000195493 Cryptophyta Species 0.000 title claims abstract description 7
- 239000003112 inhibitor Substances 0.000 title claims abstract description 5
- 150000001413 amino acids Chemical class 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 10
- 230000004069 differentiation Effects 0.000 abstract description 7
- 238000000034 method Methods 0.000 abstract description 5
- 230000008827 biological function Effects 0.000 abstract description 3
- 108050006400 Cyclin Proteins 0.000 abstract description 2
- 230000006820 DNA synthesis Effects 0.000 abstract description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 abstract description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 abstract description 2
- 206010020649 Hyperkeratosis Diseases 0.000 abstract description 2
- 208000001126 Keratosis Diseases 0.000 abstract description 2
- 102000009339 Proliferating Cell Nuclear Antigen Human genes 0.000 abstract description 2
- 108091023040 Transcription factor Proteins 0.000 abstract description 2
- 102000040945 Transcription factor Human genes 0.000 abstract description 2
- 230000004663 cell proliferation Effects 0.000 abstract description 2
- 230000036566 epidermal hyperplasia Effects 0.000 abstract description 2
- 206010020718 hyperplasia Diseases 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 238000010606 normalization Methods 0.000 abstract 1
- 241000192700 Cyanobacteria Species 0.000 description 32
- 210000001990 heterocyst Anatomy 0.000 description 21
- 235000020995 raw meat Nutrition 0.000 description 17
- 241000251468 Actinopterygii Species 0.000 description 16
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- 230000012010 growth Effects 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 101150008896 hetR gene Proteins 0.000 description 4
- 101150100573 ntcA gene Proteins 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 230000005791 algae growth Effects 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 230000004543 DNA replication Effects 0.000 description 1
- 108010081996 Photosystem I Protein Complex Proteins 0.000 description 1
- 238000010806 PrimeScriptTM RT Reagent kit Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 101150044508 key gene Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 1
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
- A01N37/46—N-acyl derivatives
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了N15多肽在制备蓝藻抑制剂中的用途，N15多肽是一个具有15个氨基酸的短肽，它具有多种生物学功能，包括调节NF‑κB转录因子结合目的基因的能力，影响NF‑κB目的基因产物蛋白质的转录和表达；阻断PCNA结合DNA聚合酶，从而阻止细胞DNA合成抑制细胞增殖；抑制上皮细胞增生，调节表皮增生及分化，使皮肤角化过程正常化等，本发明阐述了它新的生物学功能，具有较好的实用性和产业化前景。
Description
技术领域
本发明涉及生物技术领域，具体指N15多肽在制备蓝藻抑制剂中的用途。
背景技术
蓝藻之所以又称蓝细菌，是因为蓝藻是最原始、最古老的藻类. 其结构简单，无典型的细胞核，所以又称蓝细菌，属于原核生物。已知蓝藻约2000种，中国已有记录的约900种。蓝藻有极大的适应性，分布很广。在一些营养丰富的水体中，有些蓝藻常于夏季大量繁殖，并在水面形成一层蓝绿色而有腥臭味的浮沫，称为“水华”，大规模的蓝藻爆发，被称为“绿潮”(和海洋发生的赤潮对应)。绿潮引起水质恶化，严重时耗尽水中氧气而造成鱼类的死亡。
发明内容
本发明的目的在于提供一种多肽，该多肽能够抑制蓝藻生长，具有在制备细菌抑制剂中的用途，它已被现有技术公开，名为N15 多肽。
N15多肽是一个具有15个氨基酸的短肽，其序列为甲硫氨酸- 脯氨酸-酪氨酸-丝氨酸-苏氨酸-谷氨酸-亮氨酸-异亮氨酸-苯丙氨酸 -酪氨酸-异亮氨酸-谷氨酸-甲硫氨酸-天门冬氨酸-脯氨酸，它具有多种生物学功能，包括调节NF-κB转录因子结合目的基因的能力，影响NF-κB目的基因产物蛋白质的转录和表达；阻断PCNA结合 DNA聚合酶，从而阻止细胞DNA合成抑制细胞增殖；抑制上皮细胞增生，调节表皮增生及分化，使皮肤角化过程正常化等。
发明人发现在研究中发现N15可以抑制革兰氏阴性菌(大肠杆菌)、蓝细菌的生长，N15多肽能够对各种大肠杆菌和蓝藻的生长产生抑制作用。N15多肽抑制蓝藻增殖可能是通过抑制DNA复制和异形胞分化基因的表达，起到抑制蓝藻增殖的。
本发明还提供了一种N15多肽制备的抗蓝藻药物，为N15多肽的水溶液，浓度大于等于100μg/mL。
本发明的有益效果：N15多肽是一种结构清晰，易于工业化生产的多肽，它可以抑制蓝藻的增殖，效果好且无毒副作用。
附图说明
图1为N15对鱼腥蓝细菌PCC 7120的抑菌曲线。
图2为缺氮诱导48小时对照组和N15实验组鱼腥蓝细菌PCC 7120异形胞的分化情况。
图3为荧光定量RT-PCR检测N15对蓝藻hetR基因mRNA表达量的影响。
图4为荧光定量RT-PCR检测N15对蓝藻ntcA基因mRNA表达量的影响。
具体实施方式
下面结合附图和具体实验例对本发明作进一步的论证说明，以下实验例仅是对本发明的解释而本发明并不局限于以下实验例。
实验例1：N15抑制了鱼腥蓝细菌的生长
实验方法：
将鱼腥蓝细菌PCC 7120在液体BG11培养基中培养至OD750 (波长750nm处的吸光度)在0.3～0.4左右，添加N15至终浓度为 100μg/mL，同时设置不添加N15的菌株作为对照。每隔24小时用分光光度计测定菌液的OD750，连续测定8天后根据数据绘制N15 的抑菌曲线，结果如图1所示。
实验结果表明，加入N15多肽至终浓度为100ug/mL的鱼腥蓝细菌PCC 7120的生长和不加入N15的对照菌株比较，受N15多肽抑制鱼腥蓝细菌的生长较为缓慢，加入N15对鱼腥蓝细菌PCC 7120 的生长具有抑制作用。
实验例2：N15推迟了鱼腥蓝细菌的异形胞分化时间
实验方法：
鱼腥蓝细菌PCC 7120是一种丝状细菌，其菌丝体能进行光合作用。在环境中缺乏氮源的条件下，会分化出具有固氮功能的异形胞。为了观察N15处理对异形胞发育的影响，本研究中在对鱼腥蓝细菌进行缺氮处理的同时，加入100ug/mL的N15，同时设置不加入N15的对照组，诱导后进行显微镜观察。
实验结果：
鱼腥蓝细菌PCC 7120的营养细胞在形成异形胞的过程中，产氧的光系统II复合物在异形胞中失活，因此在荧光显微镜下，只有营养细胞能够发红色荧光，而异形胞没有荧光。在进行缺氮诱导的同时加入100μg/mL的N15，分别在24小时、48小时、72小时、 96小时进行显微镜观察，结果显示对照组的鱼腥蓝细菌在缺氮诱导 24小时就有异形胞的产生，而加入N15的鱼腥蓝细菌直到96小时以后才有异形胞生成，缺氮诱导48小时的照片如图2所示，加入 N15的鱼腥蓝细菌PCC 7120没有异形胞产生，菌丝体的所有细胞在荧光显微镜下都发红色荧光，而对照组菌丝有异形胞产生，这些异形胞在荧光显微镜下也没有红色荧光。由此我们得知，N15推迟了鱼腥蓝细菌PCC7120的异形胞发育生成的时间。
实验例3：N15抑制了异形胞分化基因的表达
实验方法：
为了观察N15处理对异形胞发育的影响，本研究中在对鱼腥蓝细菌进行缺氮处理的同时，加入100μg/mL的N15，同时设置不加入N15的对照组，分别提取0hour、6hour、12hour、24hour的对照组和实验组，每个样品收集50mL菌体,提取RNA，并且用DNasel 去除残余的基因组DNA。利用紫外分光光度计对总RNA定量后, 使用TakaRa公司的反转录试剂盒反转录RNA合成cDNA。具体操作步骤参见产品说明书。用TakaRa公司的试剂盒PrimeScriptTM RT Reagent Kit(Perfect Real Time)进行实时荧光定量反转录PCR。具体操作参见产品说明书。分析软件使用ROCHE LightCycler480 进行分析,定量方法为相对定量法。
实验结果：
异形胞在发育过程中需要在许多基因的参与，其中hetR和ntcA 基因是异形胞发育的关键基因，与异形胞发育的多个后期基因的转录激活依赖于hetR和ntcA。我们通过实时荧光定量反转录PCR实验检测了N15对这几个异形胞分化相关基因表达的影响，实验结果如图3～4所示。
和对照组相比，N15加入6hour、12hour、24hour后，较明显的抑制了鱼腥蓝细菌中hetR和ntcA基因的表达，由此我们得知 N15对蓝藻生长的抑制是通过抑制蓝细菌中异形胞分化相关基因的表达来实现的。
Claims (2)
1.N15多肽在制备蓝藻抑制剂中的用途；所述N15多肽是一个具有15个氨基酸的短肽，其序列为甲硫氨酸-脯氨酸-酪氨酸-丝氨酸-苏氨酸-谷氨酸-亮氨酸-异亮氨酸-苯丙氨酸-酪氨酸-异亮氨酸-谷氨酸-甲硫氨酸-天门冬氨酸-脯氨酸。
2.一种N15多肽制备的抗蓝藻药物，其特征在于：所述抗蓝藻药物为N15多肽的水溶液，浓度大于等于100μg/mL；所述N15多肽是一个具有15个氨基酸的短肽，其序列为甲硫氨酸-脯氨酸-酪氨酸-丝氨酸-苏氨酸-谷氨酸-亮氨酸-异亮氨酸-苯丙氨酸-酪氨酸-异亮氨酸-谷氨酸-甲硫氨酸-天门冬氨酸-脯氨酸。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611169815.8A CN106818746B (zh) | 2016-12-16 | 2016-12-16 | N15多肽在制备蓝藻抑制剂中的用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611169815.8A CN106818746B (zh) | 2016-12-16 | 2016-12-16 | N15多肽在制备蓝藻抑制剂中的用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106818746A CN106818746A (zh) | 2017-06-13 |
| CN106818746B true CN106818746B (zh) | 2019-08-20 |
Family
ID=59140921
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611169815.8A Active CN106818746B (zh) | 2016-12-16 | 2016-12-16 | N15多肽在制备蓝藻抑制剂中的用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106818746B (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107399874A (zh) * | 2017-07-01 | 2017-11-28 | 宁波市鄞州区渔业技术管理服务站（宁波市鄞州区海洋与渔业环境监测站、鄞州区海洋资源综合管理中心） | 一种集约化养殖尾水处理系统 |
| CN111803619A (zh) * | 2020-07-26 | 2020-10-23 | 武汉益承生物科技有限公司 | 多肽在制备创伤治疗药物中的用途 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102321158A (zh) * | 2011-08-23 | 2012-01-18 | 常州德健生物科技有限公司 | 阻止细胞dna合成抑制细胞增殖的多肽及用途 |
| CN104043102A (zh) * | 2014-03-28 | 2014-09-17 | 武汉益承生物科技股份有限公司 | 一种多肽在制备治疗核因子-κB异常活化疾病药物中的用途 |
-
2016
- 2016-12-16 CN CN201611169815.8A patent/CN106818746B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102321158A (zh) * | 2011-08-23 | 2012-01-18 | 常州德健生物科技有限公司 | 阻止细胞dna合成抑制细胞增殖的多肽及用途 |
| CN104043102A (zh) * | 2014-03-28 | 2014-09-17 | 武汉益承生物科技股份有限公司 | 一种多肽在制备治疗核因子-κB异常活化疾病药物中的用途 |
Non-Patent Citations (2)
| Title |
|---|
| 一株溶藻细菌的溶藻特性及其鉴定;裴海燕 等;《中国环境科学》;20050620;第25卷(第03期);第283-287页 |
| 抗菌肽的功能、研发与应用;单安山 等;《中国农业科学》;20120601;第45卷(第11期);第2250页第2栏倒数第1-3段、第2251页第1栏 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106818746A (zh) | 2017-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pham et al. | Cultivation of unculturable soil bacteria | |
| Lacroix et al. | Fermentation technologies for the production of probiotics with high viability and functionality | |
| Zhao et al. | Genome shuffling of Bacillus amyloliquefaciens for improving antimicrobial lipopeptide production and an analysis of relative gene expression using FQ RT-PCR | |
| US9416347B2 (en) | Method of treating bacterial contamination in a microalgae culture with pH shock | |
| CN106520866B (zh) | 通过改造糖多孢红霉菌sace_3980基因提高红霉素产量的方法 | |
| CN106818746B (zh) | N15多肽在制备蓝藻抑制剂中的用途 | |
| Kirkham et al. | A role for the cell-wall protein silacidin in cell size of the diatom Thalassiosira pseudonana | |
| CN108977890A (zh) | 一种启动子库以及利用其在细菌中构建不同强度表达系统的方法 | |
| Hu et al. | Divergent metabolic and transcriptomic responses of Synechocystis sp. PCC 6803 to salt stress after adaptive laboratory evolution | |
| CN102286614B (zh) | 调控酸胁迫抑制维生素c发酵菌株产酸的基因 | |
| Peña et al. | Genome-wide overexpression screen for sodium acetate resistance in Saccharomyces cerevisiae | |
| Liu et al. | Interactions between selected microalgae and microscopic propagules of Ulva prolifera | |
| Tong et al. | Transcriptomic analysis of morphology regulatory mechanisms of microparticles to Paraisaria dubia in submerged fermentation | |
| Lee et al. | An acceleration of carotenoid production and growth of Haematococcus lacustris induced by host-microbiota network interaction | |
| Liu et al. | Screening of antibiotics to obtain axenic cell cultures of a marine microalga Chrysotila roscoffensis | |
| CN102634447B (zh) | 一种微阵列透析室及利用该透析室的富集培养方法 | |
| McLoon et al. | Transcriptomic analysis of the Myxococcus xanthus FruA regulon, and comparative developmental transcriptomic analysis of two fruiting body forming species, Myxococcus xanthus and Myxococcus stipitatus | |
| US11555193B1 (en) | Gene ANSB knockout mutant of citrobacter werkmanii and application thereof | |
| Zhang et al. | Complete genome sequence of Acinetobacter baumanni J1, a quorum sensing-producing algicidal bacterium, isolated from Eastern Pacific Ocean | |
| CN106729609B (zh) | N15多肽在制备细菌抑制剂中的用途 | |
| CN109321618A (zh) | 一种通过糖多孢红霉菌sace_5717基因提高红霉素产量的方法 | |
| CN103014166A (zh) | 一种植物乳杆菌st-ⅲ耐盐基因的筛选方法 | |
| Zang et al. | Increasing lactose concentration is a strategy to improve the proliferation of Lactobacillus helveticus in milk | |
| Zachleder | The effect of hydroxyurea and fluorodeoxyuridine on cell cycle events in the chlorococcal alga Scenedesmus quadricauda (Chlorophyta) | |
| Bhattacharjya et al. | Depiction of growth specific changes in concentration of storage products in centric marine diatom Chaetoceros gracilis |
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 | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240426 Address after: No. 87-3 Dingshan Middle Road, Haicang District, Xiamen City, Fujian Province, 361000 Patentee after: Yicheng Kangtai (Xiamen) Biotechnology Co.,Ltd. Country or region after: China Address before: 430068 No. 28 Nanli Road, Hongshan District, Wuhan City, Hubei Province Patentee before: HUBEI University OF TECHNOLOGY Country or region before: China |
|
| TR01 | Transfer of patent right |