CN109971678A - 一种革兰氏阴性菌株及其应用 - Google Patents

一种革兰氏阴性菌株及其应用 Download PDF

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CN109971678A
CN109971678A CN201910237917.6A CN201910237917A CN109971678A CN 109971678 A CN109971678 A CN 109971678A CN 201910237917 A CN201910237917 A CN 201910237917A CN 109971678 A CN109971678 A CN 109971678A
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谢云轩
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Tianjin University
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Abstract

本发明公开了一种革兰氏阴性菌株及其应用，该菌株具有良好的抗生素生产能力，能够在低成本的高碳发酵条件下一次性获得多种可以提供成药先导化合物参考的代谢物。

Description

一种革兰氏阴性菌株及其应用
技术领域
本发明涉及一种革兰氏阴性菌株及其应用。
背景技术
微生物来源的天然药物及其衍生物在对抗微生物感染中占据着很大的比 例。微生物在长期进化中保留下来的抗生素具有很强的靶位点特异性和很高的 生物活性。近年来，抗生素的普遍使用在全球范围内导致一些具有抗生素抗药 性的人类致病菌的产生，给临床治疗带来了新的挑战。而新批准的抗生素大部 分具有与原有抗生素相似的骨架结构，极易造成靶向致病菌再次培育完善的抗 药机制以及抗生素的二次失效。较有名的实例包括ceftobiprole、daptomycin、 tigecycline这些天然抗生素药物在对多重抗药性的金黄色葡萄球菌S.aureus 和多重抗药性的肠球菌Enterococci治疗中的逐渐失效。耐药型致病菌株的快 速传播，已经导致临床上出现能对抗多种抗生素的“超级细菌”，给病人带来 了致命威胁。而拥有13亿人口的中国是世界上滥用抗生素最为严重的国家之一， 细菌整体耐药率远远高于发达国家。针对耐药型致病菌株来研发新的抗生素目 前变得刻不容缓。然而在全球抗生素资源短缺的同时，很多国际知名的制药行 业在近些年却逐渐减少研发新型抗生素的投资，造成了抗生素供求关系矛盾的 加剧。哈佛大学医学院著名天然产物学家Christopher T.Walsh教授指出，如 果不增加抗生素的研发力度，人类对抗致病菌感染的能力将会迅速退化到抗生 素发明之前的水平。充分挖掘已有的抗生素主要来源菌细胞内的次生代谢物资 源、开发全新的抗生素菌种资源迫在眉睫。
发明内容
为了解决上述技术问题，本发明的目的是提供一种革兰氏阴性菌株，具有 良好的抗生素生产能力，能够在低成本的高碳发酵条件下一次性获得多种可以 提供成药先导化合物参考的代谢物。
为解决上述技术问题，本发明采用如下技术方案：
一种革兰氏阴性菌株及其应用，具有SEQ ID NO:1所示的核苷酸序列。
进一步的，具有SEQ ID NO:1所示的核苷酸序列经修饰、取代、缺失或添 加一个或多个碱基获得的核苷酸序列。
进一步的，与SEQ ID NO:1所示的核苷酸序列具有至少80％同源性的序列。
进一步的，或者与上述序列互补的序列。
还提供了该菌株在发酵条件下获得多种可以提供成药先导化合物参考的代 谢物的应用。
与现有技术相比，本发明的有益技术效果：
该简单易得的菌株在低成本的高碳发酵条件下一次性获得多种可以提供成 药先导化合物参考的代谢物，具有良好的抗生素生产能力，大大拓展了曾被人 们低估的革兰氏阴性细菌的天然药物合成潜力，为天然药物化学提供了除传统 抗生素生产菌株之外的全新的选择。
附图说明
下面结合附图说明对本发明作进一步说明。
图1为菌株抗生素生产能力示意图。
具体实施方式
采集渤海沿岸石油烃污染土壤以期能够获取嗜麦芽窄食单胞菌。 通过松弛培养、抗生素和选择性培养基筛选，获得一系列嗜麦芽窄食 单胞菌。对分离所得菌株进行初步遗传鉴定，确认具有黄单胞菌株16S rRNA特异性扩增片段以及嗜麦芽窄食单胞菌所特有的跨膜转载蛋白 smeD基因。
对该菌株进行454测序分析，得到16S rRNA组成具有如SEQ ID NO:1所示的 核苷酸序列。
松弛培养：1％氯化钠，1％胰蛋白胨，0.5％酵母提取物；抗生素：氨苄青霉 素、卡那霉素、放线菌酮；菌株滑行能力：用于判断二者之间是否存在相关性， 如何关联。代谢物分析表明在菌株滑行期间，代谢物产量在痕量以下。抗生素 生产能力见图1。
采取1-8％高浓度小分子碳源(不超过6个碳原子)和0.1-0.8％中等浓度微生 物来源的氮源进行菌株的发酵，直接萃取菌体内的代谢产物，进行化学成分组 成及生物活性分析，得到约15种中等到低极性的代谢物成分(10-20分钟， 55％-100％乙腈洗脱)，检测到抗金黄色葡萄球菌的生物活性。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范 围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发 明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护 范围内。
SEQUENCE LISTING
<110> 天津大学
<120> 一种革兰氏阴性菌株及其应用
<130> 2018.10.11
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1266
<212> RNA
<213> Ochrobacterum anthropi
<400> 1
gtgcggtgca ccaggcgacg atccgtagct ggtctgagag gatgatcagc cacactggaa 60
ctgagacacg gtccagactc ctacgggagg cagcagtggg gaatattgga caatgggcgc 120
aagcctgatc cagccatacc gcgtgggtga agaaggcctt cgggttgtaa agcccttttg 180
ttgggaaaga aatccagccg gctaatacct ggttgggatg acggtaccca aagaataagc 240
accggctaac ttcgtgccag cagccgcggt aatacgaagg gtgcaagcgt tactcggaat 300
tactgggcgt aaagcgtgcg taggtggtcg tttaagtccg ttgtgaaagc cctgggctca 360
acctgggaac tgcagtggat actgggcgac tagagtgtgg tagagggtag cggaattcct 420
ggtgtagcag tgaaatgcgt agagatcagg aggaacatcc atggcgaagg cagctacctg 480
gaccaacact gacactgagg cacgaaagcg tggggagcaa acaggattag ataccctggt 540
agtccacgcc ctaaacgatg cgaactggat gttgggtgca atttggcacg cagtatcgaa 600
gctaacgcgt taagttcgcc gcctggggag tacggtcgca agactgaaac tcaaaggaat 660
tgacgggggc ccgcacaagc ggtggagtat gtggtttaat tcgatgcaac gcgaagaacc 720
ttacctggcc ttgacatgtc gagaactttc cagagatgga ttggtgcctt cgggaactcg 780
aacacaggtg ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc 840
aacgagcgca acccttgtcc ttagttgcca gcacgtaatg gtgggaactc taaggagacc 900
gccggtgaca aaccggagga gggtggggga tgacgtcaag tcatcatggc ccttacggcc 960
agggctacac acgtactaca atggtaggga cagagggctg caagccggcg acggtaagcc 1020
aatcccagaa accctatctc agtcccggat tggagtctgc aactcgactc catgaagtcg 1080
gaatcgctag tatcgcagat cagcattgct gcgtgaatac gttcccgact gtacacacgc 1140
ccgtcacacc atgggagttt gtgcacagag cagtagctta accttccgag tccttgcacg 1200
ggtgcgatga ctggggtgat cgtacagtag cgtatcgagt gcagcgggga cactcctact 1260
agatag 1266

Claims (4)

1.一种革兰氏阴性菌株，其特征在于：具有SEQ ID NO:1所示的核苷酸序列。
2.根据权利要求1所述的革兰氏阴性菌株，其特征在于：具有SEQ ID NO:1所示的核苷酸序列经修饰、取代、缺失或添加一个或多个碱基获得的核苷酸序列。
3.根据权利要求1所述的革兰氏阴性菌株，其特征在于：与SEQ ID NO:1所示的核苷酸序列具有至少80％同源性的序列。
4.根据权利要求1-3任意一项所述的革兰氏阴性菌株在发酵条件下获得多种可以提供成药先导化合物参考的代谢物的应用。
CN201910237917.6A 2019-03-27 2019-03-27 一种革兰氏阴性菌株及其应用 Pending CN109971678A (zh)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106749561A (zh) * 2017-01-23 2017-05-31 中国人民解放军第307医院 一种嗜麦芽窄食单胞菌外膜蛋白及其应用
CN107686491A (zh) * 2017-09-28 2018-02-13 遵义医学院 一种嗜麦芽窄食单胞菌制备的生物碱类化合物及其制备方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106749561A (zh) * 2017-01-23 2017-05-31 中国人民解放军第307医院 一种嗜麦芽窄食单胞菌外膜蛋白及其应用
CN107686491A (zh) * 2017-09-28 2018-02-13 遵义医学院 一种嗜麦芽窄食单胞菌制备的生物碱类化合物及其制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SHENG-YAN QIAN等: "New pyrazinoquinazoline alkaloids Isolated from a culture of Stenotrophomonas maltophilia QB-77", 《NATURAL PRODUCT RESEARCH》 *

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Application publication date: 20190705