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[1]张冬寒 曹明明 李延清 王绍琛 郝 鹏 冯治洋**.基于序列筛选发掘宏基因组文库中的新颖卤化酶基因*[J].应用与环境生物学报,2018,24(06):1-8.[doi:10.19675/j.cnki.1006-687x.2017.12031]
 ZHANG Donghan,CAO Mingming,LI Yanqing,et al.Discovery of novel halogenase genes using sequence-driven metagenomics approach *[J].Chinese Journal of Applied & Environmental Biology,2018,24(06):1-8.[doi:10.19675/j.cnki.1006-687x.2017.12031]
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基于序列筛选发掘宏基因组文库中的新颖卤化酶基因*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
24卷
期数:
2018年06期
页码:
1-8
栏目:
研究论文
出版日期:
2018-12-25

文章信息/Info

Title:
Discovery of novel halogenase genes using sequence-driven metagenomics approach *
文章编号:
201712031
作者:
张冬寒 曹明明 李延清 王绍琛 郝 鹏 冯治洋**
南京农业大学食品科技学院 南京 210095
Author(s):
ZHANG Donghan CAO Mingming LI Yanqing WANG Shaochen HAO Peng&FENG Zhiyang**
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
关键词:
宏基因组学序列筛选土壤微生物卤化酶基因天然卤化物
Keywords:
metagenomics sequence-driven screening soil microorganism halogenase natural halide
DOI:
10.19675/j.cnki.1006-687x.2017.12031
摘要:
绝大部分微生物的不可培养性使微生物的开发利用受到了限制,而宏基因组学策略为研究土壤中的不可培养微生物提供了途径。天然卤化物具有良好的生物活性,如抗菌活性及抗肿瘤活性等,卤化酶在催化化合物的卤化过程中,对化合物活性产生重要影响,而以卤化酶基因为探针,可以发现与之偶联的天然生物合成基因簇,为卤化物的发现提供基础。本研究利用卤化酶基因序列的保守区域设计简并引物,筛选土壤宏基因组文库获得卤化酶阳性克隆,并对获得的卤化酶基因间的进化及其与天然产物合成的关系进行了分析。结果显示:通过同源序列筛选获得了65个卤化酶阳性克隆,序列同源分析表明约85%的阳性克隆中的卤化酶基因序列与已知卤化酶的相似性低,所获卤化酶基因具有较好的新颖性和多样性。而对所获克隆中生物合成相关基因的分析表明聚酮合酶基因与卤化酶基因存在于同一克隆中,其可能与卤代I型聚酮合成相关。本研究基于序列筛选的方法,从宏基因组文库中发现了新的卤化酶基因和聚酮合酶基因,为进一步发现新颖的天然卤化物生物合成基因簇及天然卤化物奠定了基础。(表1图3参29)
Abstract:
It is of great significan ce to obtain new natural products with diverse activities through the study of soil microorganisms . However, less than 1% of the total soil microorganisms could be cultured under the laboratory conditions limiting the new compounds discovery. Metagenomics, by which the genomic DNA of soil microorganisms could be extracted and expressed in heterologous host , provides a new approach for functional study of soil microorganisms. Natural halides have good bioactivities, such as antibacterial activity and antitumor activity. Halogenases play an important role in biosynthesis and introducing bioactivities of halogenated compounds . To investigate the potential of halogenated compound production of soil microorganisms, a soil metagenomic library was screened by PCR for clones which harbor reduced flavin adenine dinucleotide (FADH 2) - dependent halogenase genes. Sixty-five positive clones were identified from the library and the amino acid sequences of halogenase genes within the positive clones were analyzed. Phylogenetic analyses r evealed that more than 85% of them were separated from known halogenases to form new clades in the phylogen etic tree and the soil-derived halogenases showed high diversity . New type I polyketide biosynthetic gene sequence was identified by further biosynthetic gene analysis of the positive clones , which probably was related to the biosynthesis of halogenated type I polyketide. In conclusion, n ovel and diverse halogenase genes were identified on sixty-five metagenomic clones using sequence-driven metagenomic approach, which laid a foundation for further discovery of novel natural halides biosynthetic gene clusters and novel halides.

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备注/Memo

备注/Memo:
收稿日期: 2017-12-12 接受日期: 2018-03-06
*国家自然科学基金项目(31370088, 31500035)资助
**通讯作者(E-mail:zfeng@njau.edu.cn)
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更新日期/Last Update: 2018-06-05