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微生物学报

一株降解PET单体对苯二甲酸的施氏假单胞菌的筛选和基因组分析
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国家自然科学基金(31961133014);国家重点研发计划(2019YFA0706900)


Screening and genome analysis of a Pseudomonas stutzeri that degrades PET monomer terephthalate
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    摘要:

    【目的】筛选和鉴定能够降解聚对苯二甲酸乙二醇酯(PET)单体的微生物,并分析代谢途径。【方法】从青岛小涧西固体废弃物综合处置厂采集样品,以PET单体对苯二甲酸为唯一碳源筛选获得能够代谢对苯二甲酸的菌株TPA3;16S rRNA序列分析确定TPA3菌株的分类地位;采用二代和三代高通量测序技术进行基因组de novo测序和分析;通过氨基酸序列比对分析TPA3菌株代谢对苯二甲酸和乙二醇的途径和基因;验证TPA3菌株的可遗传操作性。【结果】TPA3菌株被鉴定为施氏假单胞菌(Pseudomonas stutzeri);它能够在30℃、220 r/min培养条件下,在30 h内消耗10.60 g/L对苯二甲酸;经过驯化培养,也能够代谢乙二醇;其基因组由1条染色体和3个质粒组成,总大小约为4.55 Mb;推测该菌株具有经典的对苯二甲酸代谢途径以及与Pseudomonas putida KT2440相似的乙二醇代谢途径;采用假单胞菌遗传操作技术可对TPA3菌株进行遗传改造。【结论】TPA3菌株能够降解PET单体对苯二甲酸和乙二醇,并具有可遗传操作性,在PET废物生物处理技术中具有潜在的应用价值。

    Abstract:

    [Objective] To screen and identify microorganisms capable of metabolizing polyethylene terephthalate monomer terephthalate and analyze the catabolic pathways.[Methods] Collected samples from Qingdao Xiaojianxi solid waste comprehensive disposal plant and screened strains capable of metabolizing terephthalate using the medium with terephthalate as the sole carbon source; 16S rRNA phylogenetic tree analysis was used to determine the taxonomic status of TPA3 strain; de novo sequencing was implemented using the second-generation and the third-generation high-throughput sequencing technologies; the terephthalate and ethylene glycol catabolic pathways and the related genes of TPA3 were analyzed through amino acid sequence alignment; the genetic manipulability of TPA3 was verified by the reported genetic manipulative techniques.[Results] TPA3 was identified as Pseudomonas stutzeri; it can metabolize 10.60 g/L terephthalate within 30 hours at 30℃. After culture and domestication, it could also catabolize ethylene glycol. The complete genome was composed of one chromosome and three plasmids for a total genome size of 4.55 Mb. It was speculated that TPA3 strain catabolized terephthalate in a classic way, and the ethylene glycol catabolic pathway should be similar to Pseudomonas putida KT2440. The genetic manipulation technology of Pseudomonas could be used for TPA3.[Conclusion] TPA3 strain could degrade polyethylene terephthalate monomers terephthalate and ethylene glycol, and could be genetically modified, showing potential application value in polyethylene terephthalate waste biological treatment technology.

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刘盼,张彤,郑义,李庆宾,梁泉峰,祁庆生. 一株降解PET单体对苯二甲酸的施氏假单胞菌的筛选和基因组分析. 微生物学报, 2022, 62(1): 200-212

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  • 收稿日期:2021-03-22
  • 最后修改日期:2021-05-15
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  • 在线发布日期: 2022-01-06
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