Enrichment and Isolation of Aerobic and Anaerobic Methanotrophs

Rhee, Sung‐Keun; Awala, Samuel Imisi; Nguyen, Ngoc-Loi

doi:10.1007/978-3-030-23261-0_2

Cited by 1 publication

(1 citation statement)

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“…Methanotrophs, or microbes that consume CH 4 , provide a biological and natural methane sink either via aerobic or anaerobic routes of CH 4 consumption ( Rhee et al, 2019 ). However, among the methanotrophs, aerobic methanotrophic bacteria have been mostly isolated in pure cultures and studied ( Guerrero-Cruz et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Optimization of electroporation method and promoter evaluation for type-1 methanotroph, Methylotuvimicrobium alcaliphilum

Goswami,

Singer,

Simmons

et al. 2024

Front. Bioeng. Biotechnol.

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Methanotrophic bacteria are promising hosts for methane bioconversion to biochemicals or bioproducts. However, due to limitations associated with long genetic manipulation timelines and, lack of choice in genetic tools required for strain engineering, methanotrophs are currently not employed for bioconversion technologies. In this study, a rapid and reproducible electroporation protocol is developed for type 1 methanotroph, Methylotuvimicrobium alcaliphilum using common laboratory solutions, analyzing optimal electroshock voltages and post-shock cell recovery time. Successful reproducibility of the developed method was achieved when different replicative plasmids were assessed on lab adapted vs. wild-type M. alcaliphilum strains (DASS vs. DSM19304). Overall, a ∼ 3-fold decrease in time is reported with use of electroporation protocol developed here, compared to conjugation, which is the traditionally employed approach. Additionally, an inducible (3-methyl benzoate) and a constitutive (sucrose phosphate synthase) promoter is characterized for their strength in driving gene expression.

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Section: Introductionmentioning

confidence: 99%