Carbon-negative production of acetone and isopropanol by gas fermentation at industrial pilot scale

Liew, FungMin; Nogle, Robert; Abdalla, Tanus; Rasor, Blake J.; Canter, Christina E.; Jensen, Rasmus O.; Wang, Lan; Strutz, Jonathan; Chirania, Payal; Tissera, Sashini De; Mueller, Alexander P.; Ruan, Zhenhua; Gao, Allan; Tran, Loan; Engle, Nancy L.; Bromley, Jason C; Daniell, James; Conrado, Robert J.; Tschaplinski, Timothy J.; Giannone, Richard J.; Hettich, Robert L.; Karim, Ashty S.; Simpson, Séan D.; Brown, Steven D.; Leang, Ching; Jewett, Michael C.; Köpke, Michael

doi:10.1038/s41587-021-01195-w

Cited by 230 publications

(183 citation statements)

References 82 publications

Supporting

Mentioning

183

Contrasting

Order By: Relevance

“…In a very recent study by Liew et al . variation of the promoter alone led to an 11-fold improvement in production when optimizing the acetone biosynthesis pathway for expression in C. autoethanogenum [ 71 ] .…”

Section: Discussionmentioning

confidence: 99%

Metabolic engineering of Clostridium ljungdahlii for the production of hexanol and butanol from CO2 and H2

2022

View full text Add to dashboard Cite

Background The replacement of fossil fuels and petrochemicals with sustainable alternatives is necessary to mitigate the effects of climate change and also to counteract diminishing fossil resources. Acetogenic microorganisms such as Clostridium spp. are promising sources of fuels and basic chemical precursors because they efficiently utilize CO and CO2 as carbon source. However the conversion into high titers of butanol and hexanol is challenging. Results Using a metabolic engineering approach we transferred a 17.9-kb gene cluster via conjugation, containing 13 genes from C. kluyveri and C. acetobutylicum for butanol and hexanol biosynthesis, into C. ljungdahlii. Plasmid-based expression resulted in 1075 mg L−1 butanol and 133 mg L−1 hexanol from fructose in complex medium, and 174 mg L−1 butanol and 15 mg L−1 hexanol from gaseous substrate (20% CO2 and 80% H2) in minimal medium. Product formation was increased by the genomic integration of the heterologous gene cluster. We confirmed the expression of all 13 enzymes by targeted proteomics and identified potential rate-limiting steps. Then, we removed the first-round selection marker using CRISPR/Cas9 and integrated an additional 7.8 kb gene cluster comprising 6 genes from C. carboxidivorans. This led to a significant increase in the hexanol titer (251 mg L−1) at the expense of butanol (158 mg L−1), when grown on CO2 and H2 in serum bottles. Fermentation of this strain at 2-L scale produced 109 mg L−1 butanol and 393 mg L−1 hexanol. Conclusions We thus confirmed the function of the butanol/hexanol biosynthesis genes and achieved hexanol biosynthesis in the syngas-fermenting species C. ljungdahlii for the first time, reaching the levels produced naturally by C. carboxidivorans. The genomic integration strain produced hexanol without selection and is therefore suitable for continuous fermentation processes.

show abstract

Section: Discussionmentioning

confidence: 99%

Metabolic engineering of Clostridium ljungdahlii for the production of hexanol and butanol from CO2 and H2

2022

View full text Add to dashboard Cite

show abstract

“…For example, iPROBE allowed us to screen 54 different linear pathways for 3-hydroxybutyrate production, 205 variants of a butanol pathway, and 580 unique linear pathway combinations for limonene synthesis starting with glucose as a carbon source [13][14][15][16][17] . iPROBE additionally enabled the optimization of 15 competing reactions for acetone biosynthesis in C. autoethanogenum 18 and pathway performance in each study correlated well with in vivo yields. Adapting iPROBE to study cyclic and iterative pathways like r-BOX could allow engineering product specificity but has not previously been attempted.…”

Section: Introductionmentioning

confidence: 91%

Cell-free prototyping enables implementation of optimized reverse β-oxidation pathways in heterotrophic and autotrophic bacteria

Vögeli

Schulz

Garg

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Carbon-negative synthesis of biochemical products has the potential to mitigate global CO2 emissions. An attractive route to do this is the reverse β-oxidation (r-BOX) pathway coupled to the Wood-Ljungdahl pathway. Here, we optimized and implemented r-BOX for the synthesis of C4-C6 acids and alcohols. With a high-throughput in vitro prototyping workflow, we screened 762 unique pathway combinations using cell-free extracts tailored for r-BOX to identify enzyme sets for enhanced product selectivity. Implementation of these pathways into Escherichia coli generated designer strains for the selective production of butanoic acid (4.9 +/- 0.1 gL-1), hexanoic acid (3.06 +/- 0.03 gL-1) and 1-hexanol (1.0 +/- 0.1 gL-1) at the best performance reported to date in this bacterium. We also generated Clostridium autoethanogenum strains able to produce 1-hexanol from syngas, achieving a titer of 0.26 gL-1 in a 1.5-L continuous fermentation. Our strategy enables optimization of rBOX derived products for biomanufacturing and industrial biotechnology.

show abstract

“…After the acetate production, acetate will be further upgraded using a heterotroph microalgae strain into lipids (high-value omega-3 fatty acids, e.g., docosahexaenoic acid or DHA, and fatty acids for biodiesel production) ( https://iocl.com/NewsDetails/58912 ). Very recently, LanzaTech published the study that converts CO 2 to acetone and isopropanol at a rate of ∼3 g/L/h at industrial pilot scale ( Liew et al, 2022 ). Also, CO 2 has been used to produce starch in the form of amylose and amylopectin in a cell-free system ( Cai et al, 2021 ).…”

Section: Diversifying Value-added Products In Biotechnology Industrymentioning

confidence: 99%

Microbial Utilization of Next-Generation Feedstocks for the Biomanufacturing of Value-Added Chemicals and Food Ingredients

Zhang

Ottenheim

Weingarten

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Global shift to sustainability has driven the exploration of alternative feedstocks beyond sugars for biomanufacturing. Recently, C1 (CO2, CO, methane, formate and methanol) and C2 (acetate and ethanol) substrates are drawing great attention due to their natural abundance and low production cost. The advances in metabolic engineering, synthetic biology and industrial process design have greatly enhanced the efficiency that microbes use these next-generation feedstocks. The metabolic pathways to use C1 and C2 feedstocks have been introduced or enhanced into industrial workhorses, such as Escherichia coli and yeasts, by genetic rewiring and laboratory evolution strategies. Furthermore, microbes are engineered to convert these low-cost feedstocks to various high-value products, ranging from food ingredients to chemicals. This review highlights the recent development in metabolic engineering, the challenges in strain engineering and bioprocess design, and the perspectives of microbial utilization of C1 and C2 feedstocks for the biomanufacturing of value-added products.

show abstract

Carbon-negative production of acetone and isopropanol by gas fermentation at industrial pilot scale

Cited by 230 publications

References 82 publications

Metabolic engineering of Clostridium ljungdahlii for the production of hexanol and butanol from CO2 and H2

Metabolic engineering of Clostridium ljungdahlii for the production of hexanol and butanol from CO2 and H2

Cell-free prototyping enables implementation of optimized reverse β-oxidation pathways in heterotrophic and autotrophic bacteria

Microbial Utilization of Next-Generation Feedstocks for the Biomanufacturing of Value-Added Chemicals and Food Ingredients

Contact Info

Product

Resources

About