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- W4250902087 abstract "Abstract Background: Industrial biofuels and other value-added products can be produced from metabolically engineered microorganisms. Methylomonas sp. DH-1 is a candidate platform for bioconversion that uses methane as a carbon source. Although several genetic engineering techniques have been developed to work with Methylomonas sp. DH-1, the genetic manipulation of plasmids remains difficult because of the restriction-modification (RM) system present in the bacteria. Therefore, the RM system in Methylomonas sp. DH-1 must be identified to improve the genetic engineering prospects of this microorganism. Results: We identified a DNA methylation site, TGGCCA, and its corresponding cytosine methyltransferase for the first time in Methylomonas sp. DH-1 through whole-genome bisulfite sequencing. The methyltransferase was confirmed to methylate the fourth nucleotide of TGG C CA. In general, methylated plasmids exhibited better transformation efficiency under the protection of the RM system than non-methylated plasmids did. As expected, when we transformed Methylomonas sp. DH-1 with plasmid DNA harboring the psy gene, the metabolic flux towards carotenoid increased. The methyltransferase-treated plasmid exhibited an increase in transformation efficiency of 2.5 × 10 3 CFU/μg (124 %). The introduced gene increased the production of carotenoid by 26 %. In addition, the methyltransferase-treated plasmid harboring anti- psy sRNA gene exhibited an increase in transformation efficiency by 70 % as well. The production of carotenoid was decreased by 40 % when the psy gene was translationally repressed by anti- psy sRNA. Conclusions: Plasmid DNA methylated by the discovered cytosine methyltransferase from Methylomonas sp. DH-1 had a higher transformation efficiency than non-treated plasmid DNA. The RM system identified in this study may facilitate the plasmid-based genetic manipulation of methanotrophs." @default.
- W4250902087 created "2022-05-12" @default.
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- W4250902087 date "2020-11-18" @default.
- W4250902087 modified "2023-09-27" @default.
- W4250902087 title "Identification of a cytosine methyltransferase that improves transformation efficiency in Methylomonas sp. DH-1" @default.
- W4250902087 doi "https://doi.org/10.21203/rs.3.rs-38183/v2" @default.
- W4250902087 hasPublicationYear "2020" @default.
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