ArticleBiotechnology for biofuels2015
A novel arabinose-inducible genetic operation system developed for Clostridium cellulolyticum.
Article in Biotechnology for biofuels, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 31 citations in OpenAlex.
- Digital PCR: a tool in clostridial mutant selection and detection.Applied microbiology and biotechnology · 2023Article
- Development of an efficient ClosTron system for gene disruption in Ruminiclostridium papyrosolvens.Applied microbiology and biotechnology · 2023Article
- Genetic sensor-regulators functional in Clostridia.Journal of industrial microbiology & biotechnology · 2020Article
- Impact of ammonium sulfite-based sequential pretreatment combinations on two distinct saccharifications of wheat straw.RSC advances · 2020Article
- Synthetic Biology Tools for Genome and Transcriptome Engineering of SolventogenicFrontiers in bioengineering and biotechnology · 2020Review
- Construction of consolidated bio-saccharification biocatalyst and process optimization for highly efficient lignocellulose solubilization.Biotechnology for biofuels · 2019Article
- Recent Developments of the Synthetic Biology Toolkit forFrontiers in microbiology · 2018Review
- Determination of the native features of the exoglucanase Cel48S fromBiotechnology for biofuels · 2018Article
- Efficient whole-cell-catalyzing cellulose saccharification using engineeredBiotechnology for biofuels · 2017Article
- Dual control system - A novel scaffolding architecture of an inducible regulatory device for the precise regulation of gene expression.Metabolic engineering · 2016Article
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Authors and funding
4 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundClostridium cellulolyticum and other cellulolytic Clostridium strains are natural producers of lignocellulosic biofuels and chemicals via the consolidated bioprocessing (CBP) route, and systems metabolic engineering is indispensable to meet the cost-efficient demands of industry. Several genetic tools have been developed for Clostridium strains, and an efficient and stringent inducible genetic operation system is still required for the precise regulation of the target gene function.
resultsHere, we provide a stringent arabinose-inducible genetic operation (ARAi) system for C. cellulolyticum, including an effective gene expression platform with an oxygen-independent fluorescent reporter, a sensitive MazF-based counterselection genetic marker, and a precise gene knock-out method based on an inducible ClosTron system. A novel arabinose-inducible promoter derived from Clostridium acetobutylicum is employed in the ARAi system to control the expression of the target gene, and the gene expression can be up-regulated over 800-fold with highly induced stringency. The inducible ClosTron method of the ARAi system decreases the off-target frequency from 100% to 0, which shows the precise gene targeting in C. cellulolyticum. The inducible effect of the ARAi system is specific to a universal carbon source L-arabinose, implying that the system could be used widely for clostridial strains with various natural substrates.
conclusionsThe inducible genetic operation system ARAi developed in this study, containing both controllable gene expression and disruption tools, has the highest inducing activity and stringency in Clostridium by far. Thus, the ARAi system will greatly support the efficient metabolic engineering of C. cellulolyticum and other mesophilic Clostridium strains for lignocellulose bioconversion.
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