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[1]张冬寒,曹明明,李延清,等.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):1301-1306.[doi:10.19675/j.cnki.1006-687x.2017.12031]
 ZHANG Donghan,et al..Discovery of novel halogenase genes using sequence-driven metagenomics approach[J].Chinese Journal of Applied & Environmental Biology,2018,24(06):1301-1306.[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:
1301-1306
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
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 et al.
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
分类号:
Q78 : S154.39
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. 天然卤化物具有抗菌活性和抗肿瘤活性等生物活性,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. 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其中一个克隆中同时存在聚酮合酶基因与卤化酶基因,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. (图3表1参29)
Abstract:
It is of great significance to ability to obtain new natural products with diverse activities through the study of soil microorganisms. However, less than 1% of the total soil microorganisms can be cultured under laboratory conditions, thus limiting the discovery of new compounds. Metagenomics, by which the genomic DNA of soil microorganisms can be extracted and expressed in heterologous hosts, provides a new approach for the functional study of soil microorganisms. Natural halides have good bioactivities, including antibacterial and antitumor activities. Halogenases play an important role in biosynthesis, and introducing bioactivities of halogenated compounds. To investigate the potential of halogenated compounds production from soil microorganisms, a soil metagenomic library was screened by PCR for clones harboring reduced flavin adenine dinucleotide (FADH2) - 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 revealed that more than 85% of these genes were separated from known halogenases to form new clades in the phylogenetic tree; moreover the soil-derived halogenases showed high diversity. By further biosynthetic gene analysis of the positive clones, a new type I polyketide biosynthetic gene sequence was identified, which is probably related to the biosynthesis of the halogenated type I polyketide. In conclusion, novel and diverse halogenase genes were identified on sixty-five metagenomic clones using a sequence-driven metagenomic approach, laying a foundation for the further discovery of novel natural halides biosynthetic gene clusters and halides.

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更新日期/Last Update: 2018-12-25