Microbial production of chemicals driven by CRISPR-Cas systems

Shi, Shuobo; Qi, Nailing; Nielsen, Jens

doi:10.1016/j.copbio.2021.07.002

Cited by 22 publications

(6 citation statements)

References 50 publications

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“…Also, it is important to note that the in silico strains S2 and S3 of Synechocystis are favored due to their superior biomass and ethanol productivities, emphasizing the significance of strain engineering for optimal performance in biorefineries. In the context of current research, gene editing and precise post-transcriptional control over gene expression tools, such as CRISPR-Cas9 − and small regulatory RNAs (sRNAs), − respectively, have emerged as revolutionary techniques for cyanobacteria strain design. This combined approach offers researchers unprecedented capabilities to engineer cyanobacteria with enhanced metabolic pathways, increased productivity, and specialized adaptability to varying environmental conditions.…”

Section: Resultsmentioning

confidence: 99%

Process Systems Engineering Approach for the Sustainable Design of Cyanobacteria-Based Integrated Biorefineries and Their Heat Exchanger Network

Ramos,

Lasry Testa,

Ramos

et al. 2024

ACS Sustainable Chem. Eng.

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As climate change increasingly impacts the environment and society worldwide, advanced biofuels produced from cyanobacteria constitute a nature-based solution to the capture and storage of the atmospheric greenhouse gas carbon dioxide by continual or enhanced biological processes. We propose a mixedinteger nonlinear programming problem to address the optimal design of a biorefinery and its heat exchanger network based on Synechocystis sp. PCC 6803, aiming to maximize the sustainability net present value in a superstructure approach. The optimal scheme includes production of pigments and fourth-generation bioethanol, self-supply of energy, and recycling of enriched nutrient streams. Numerical results provide a sustainability net present value of 117.55 M USD, rendering a competitive fourth-generation bioethanol production cost of 0.40 USD/kg of bioethanol. Simultaneous heat integration improves the objective function, showing that reducing environmental burden has a greater impact than minimizing the capital investment in heat exchangers, compared to the case of no heat integration. Sensitivity analysis shows that further enhancement of Synechocystis' ethanol productivity is crucial to improve sustainability.

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Section: Resultsmentioning

confidence: 99%

Process Systems Engineering Approach for the Sustainable Design of Cyanobacteria-Based Integrated Biorefineries and Their Heat Exchanger Network

Ramos,

Lasry Testa,

Ramos

et al. 2024

ACS Sustainable Chem. Eng.

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“…As a powerful genome editing tool, the CRISPR-Cas9 system has been widely used in many organisms [33]. So far, several CRISPR systems have been developed for genome editing in B. subtilis [27,[34][35][36].…”

Section: Discussionmentioning

confidence: 99%

Development and application of a rapid all-in-one plasmid CRISPR-Cas9 system for iterative genome editing in Bacillus subtilis

et al. 2022

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Background Bacillus subtilis, an important industrial microorganism, is commonly used in the production of industrial enzymes. Genome modification is often necessary to improve the production performance of cell. The dual-plasmid CRISPR-Cas9 system suitable for iterative genome editing has been applied in Bacillus subtilis. However, it is limited by the selection of knockout genes, long editing cycle and instability. Results To address these problems, we constructed an all-in-one plasmid CRISPR-Cas9 system, which was suitable for iterative genome editing of B. subtilis. The PEG4000-assisted monomer plasmid ligation (PAMPL) method greatly improved the transformation efficiency of B. subtilis SCK6. Self-targeting sgRNArep transcription was tightly controlled by rigorous promoter PacoR, which could induce the elimination of plasmids after genome editing and prepare for next round of genome editing. Our system achieved 100% efficiency for single gene deletions and point mutations, 96% efficiency for gene insertions, and at least 90% efficiency for plasmid curing. As a proof of concept, two extracellular protease genes epr and bpr were continuously knocked out using this system, and it only took 2.5 days to complete one round of genome editing. The engineering strain was used to express Douchi fibrinolytic enzyme DFE27, and its extracellular enzyme activity reached 159.5 FU/mL. Conclusions We developed and applied a rapid all-in-one plasmid CRISPR-Cas9 system for iterative genome editing in B. subtilis, which required only one plasmid transformation and curing, and accelerated the cycle of genome editing. To the best of our knowledge, this is the rapidest iterative genome editing system for B. subtilis. We hope that the system can be used to reconstruct the B. subtilis cell factory for the production of various biological molecules.

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“…To obtain industrially applied strains, multicopy chromosomal integration of target pathway genes may be a solution. The CRISPR-Cas system allows the knock-in of heterologous genes, integration of large synthetic pathways as well as the combinatorial and multiplex modifications in chromosomes [85][86][87][88][89]. Using CRISPR-associated transposases (MUCICAT) and targeting the crRNA to multicopy loci of the E. coli genome, up to 10 copies of integration were achieved [90].…”

Section: Increasing Production Strain Stabilitymentioning

confidence: 99%

Toward bioproduction of oxo chemicals from C1 feedstocks using isobutyraldehyde as an example

Guo

Sun

Huo

2022

Biotechnol Biofuels

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Oxo chemicals are valuable chemicals for synthesizing a wide array of industrial and consumer products. However, producing of oxo chemicals is predominately through the chemical process called hydroformylation, which requires petroleum-sourced materials and generates abundant greenhouse gas. Current concerns on global climate change have renewed the interest in reducing greenhouse gas emissions and recycling the plentiful greenhouse gas. A carbon–neutral manner in this regard is producing oxo chemicals biotechnologically using greenhouse gas as C1 feedstocks. Exemplifying isobutyraldehyde, this review demonstrates the significance of using greenhouse gas for oxo chemicals production. We highlight the current state and the potential of isobutyraldehyde synthesis with a special focus on the in vivo and in vitro scheme of C1-based biomanufacturing. Specifically, perspectives and scenarios toward carbon– and nitrogen–neutral isobutyraldehyde production are proposed. In addition, key challenges and promising approaches for enhancing isobutyraldehyde bioproduction are thoroughly discussed. This study will serve as a reference case in exploring the biotechnological potential and advancing oxo chemicals production derived from C1 feedstocks.

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Microbial production of chemicals driven by CRISPR-Cas systems

Cited by 22 publications

References 50 publications

Process Systems Engineering Approach for the Sustainable Design of Cyanobacteria-Based Integrated Biorefineries and Their Heat Exchanger Network

Process Systems Engineering Approach for the Sustainable Design of Cyanobacteria-Based Integrated Biorefineries and Their Heat Exchanger Network

Development and application of a rapid all-in-one plasmid CRISPR-Cas9 system for iterative genome editing in Bacillus subtilis

Toward bioproduction of oxo chemicals from C1 feedstocks using isobutyraldehyde as an example

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