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Development of a Targeted Gene Disruption System in the Poly(Ethylene Terephthalate)-Degrading Bacterium Ideonella sakaiensis and Its Applications to PETase and MHETase Genes
http://hdl.handle.net/10061/0002000755
http://hdl.handle.net/10061/0002000755e3160e95-6d8c-4f21-96c4-301d0fdfd502
| 名前 / ファイル | ライセンス | アクション |
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fulltext (1.7 MB)
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| アイテムタイプ | 学術雑誌論文 / Journal Article(1) | |||||||||
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| 公開日 | 2025-01-31 | |||||||||
| タイトル | ||||||||||
| タイトル | Development of a Targeted Gene Disruption System in the Poly(Ethylene Terephthalate)-Degrading Bacterium Ideonella sakaiensis and Its Applications to PETase and MHETase Genes | |||||||||
| 言語 | ||||||||||
| 言語 | eng | |||||||||
| 資源タイプ | ||||||||||
| 資源タイプ | journal article | |||||||||
| アクセス権 | ||||||||||
| アクセス権 | open access | |||||||||
| 著者 |
Hachisuka Shin-ichi
× Hachisuka Shin-ichi
× Nishii Tarou
× 吉田, 昭介 |
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| 抄録 | ||||||||||
| 内容記述タイプ | Abstract | |||||||||
| 内容記述 | Poly(ethylene terephthalate) (PET) is a commonly used synthetic plastic; however, its nonbiodegradability results in a large amount of waste accumulation that has a negative impact on the environment. Recently, a PET-degrading bacterium, Ideonella sakaiensis 201-F6 strain, was isolated, and the enzymes involved in PET digestion, PET hydrolase (PETase), and mono(2-hydroxyethyl) terephthalic acid (MHET) hydrolase (MHETase) were identified. Despite the great potentials of I. sakaiensis in bioremediation and biorecycling, approaches to studying this bacterium remain limited. In this study, to enable the functional analysis of PETase and MHETase genes in vivo, we have developed a gene disruption system in I. sakaiensis. The pT18mobsacB-based disruption vector harboring directly connected 5′- and 3′-flanking regions of the target gene for homologous recombination was introduced into I. sakaiensis cells via conjugation. First, we deleted the orotidine 5′-phosphate decarboxylase gene (pyrF) from the genome of the wild-type strain, producing the ΔpyrF strain with 5-fluoroorotic acid (5-FOA) resistance. Next, using the ΔpyrF strain as a parent strain and pyrF as a counterselection marker, we disrupted the genes for PETase and MHETase. The growth of both Δpetase and Δmhetase strains on terephthalic acid (TPA; one of the PET hydrolytic products) was comparable to that of the parent strain. However, these mutant strains dramatically decreased the growth level on PET to that on a no-carbon source. Moreover, the Δpetase strain completely abolished PET degradation capacity. These results demonstrate that PETase and MHETase are essential for I. sakaiensis metabolism of PET. | |||||||||
| 書誌情報 |
en : Applied and Environmental Microbiology 巻 87, 号 18, 発行日 2021-08-26 |
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| 出版者 | ||||||||||
| 出版者 | American Society for Microbiology | |||||||||
| ISSN | ||||||||||
| 収録物識別子タイプ | EISSN | |||||||||
| 収録物識別子 | 1098-5336 | |||||||||
| 出版者版DOI | ||||||||||
| 関連タイプ | isVersionOf | |||||||||
| 識別子タイプ | DOI | |||||||||
| 関連識別子 | https://doi.org/10.1128/AEM.00020-21 | |||||||||
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| 関連タイプ | isVersionOf | |||||||||
| 識別子タイプ | URI | |||||||||
| 関連識別子 | https://journals.asm.org/doi/full/10.1128/aem.00020-21 | |||||||||
| 権利 | ||||||||||
| 権利情報 | $00A9 2021 American Society for Microbiology. | |||||||||
| 著者版フラグ | ||||||||||
| 出版タイプ | AM | |||||||||
