Isobutene production in Synechocystis sp. PCC 6803 by introducing α-ketoisocaproate dioxygenase from Rattus norvegicus

Mustila, Henna; Kugler, Amit; Stensjö, Karin

doi:10.1016/j.mec.2021.e00163

Cited by 24 publications

(21 citation statements)

References 41 publications

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“…The broad host range RSF1010 replicon-based self-replicating plasmid is widely used when demonstrating biofuel production in cyanobacteria [ 18 , 21 , 22 ]. However, this is the first study expressing genes with both a self-replicating plasmid-based system and a genome-based system in Synechocystis cells.…”

Section: Resultsmentioning

confidence: 99%

Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production

Xie

Lindblad

2022

Microb Cell Fact

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Background Cyanobacteria, photosynthetic microorganisms, are promising green cell factories for chemical production, including biofuels. Isobutanol, a four-carbon alcohol, is considered as a superior candidate as a biofuel for its high energy density with suitable chemical and physical characteristics. The unicellular cyanobacterium Synechocystis PCC 6803 has been successfully engineered for photosynthetic isobutanol production from CO2 and solar energy in a direct process. Results Heterologous expression of α-ketoisovalerate decarboxylase (KivdS286T) is sufficient for isobutanol synthesis via the 2-keto acid pathway in Synechocystis. With additional expression of acetolactate synthase (AlsS), acetohydroxy-acid isomeroreductase (IlvC), dihydroxy-acid dehydratase (IlvD), and alcohol dehydrogenase (Slr1192OP), the Synechocystis strain HX42, with a functional 2-keto acid pathway, showed enhanced isobutanol production reaching 98 mg L−1 in short-term screening experiments. Through modulating kivdS286T copy numbers as well as the composition of the 5′-region, a final Synechocystis strain HX47 with three copies of kivdS286T showed a significantly improved isobutanol production of 144 mg L−1, an 177% increase compared to the previously reported best producing strain under identical conditions. Conclusions This work demonstrates the feasibility to express heterologous genes with a combination of self-replicating plasmid-based system and genome-based system in Synechocystis cells. Obtained isobutanol-producing Synechocystis strains form the base for further investigation of continuous, long-term-photosynthetic isobutanol production from solar energy and carbon dioxide. Graphic abstract

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Section: Resultsmentioning

confidence: 99%

Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production

Xie

Lindblad

2022

Microb Cell Fact

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“…Similarly, isobutene (a gaseous fuel) production in Synechocystis sp. PCC 6803 has been enhanced by the introduction of the α-ketoisocaproate dioxygenase gene from Rattus norvegicus (RnKICD), which resulted in optimization of the isobutene pathway [ 142 ]. Also, the industrial scale-up of terpenoids through sequential heterologous expression of bottleneck enzymes of Methylerythritol 4-phosphate pathway in Synechocystis sp.…”

Section: Recent Metabolic Advances In Microorganism For Biofuel Produ...mentioning

confidence: 99%

Recent advances in metabolic engineering of microorganisms for advancing lignocellulose-derived biofuels

et al. 2022

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Combating climate change and ensuring energy supply to a rapidly growing global population has highlighted the need to replace petroleum fuels with clean, and sustainable renewable fuels. Biofuels offer a solution to safeguard energy security with reduced ecological footprint and process economics. Over the past years, lignocellulosic biomass has become the most preferred raw material for the production of biofuels, such as fuel, alcohol, biodiesel, and biohydrogen. However, the cost-effective conversion of lignocellulose into biofuels remains an unsolved challenge at the industrial scale. Recently, intensive efforts have been made in lignocellulose feedstock and microbial engineering to address this problem. By improving the biological pathways leading to the polysaccharide, lignin, and lipid biosynthesis, limited success has been achieved, and still needs to improve sustainable biofuel production. Impressive success is being achieved by the retouring metabolic pathways of different microbial hosts. Several robust phenotypes, mostly from bacteria and yeast domains, have been successfully constructed with improved substrate spectrum, product yield and sturdiness against hydrolysate toxins. Cyanobacteria is also being explored for metabolic advancement in recent years, however, it also remained underdeveloped to generate commercialized biofuels. The bacterium Escherichia coli and yeast Saccharomyces cerevisiae strains are also being engineered to have cell surfaces displaying hydrolytic enzymes, which holds much promise for near-term scale-up and biorefinery use. Looking forward, future advances to achieve economically feasible production of lignocellulosic-based biofuels with special focus on designing more efficient metabolic pathways coupled with screening, and engineering of novel enzymes.

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“…One of the key objectives of NordAqua is to explore new alternatives for sustainable large-scale photosynthetic bio-production using synthetic biology as the enabling technology (Figure 1C). To achieve this, NordAqua research focuses on the development of cyanobacterial chasses with improved photosynthesis (see above), advancing different synthetic biology toolboxes for fast and stable metabolic engineering and employing modular metabolic engineering strategies for the efficient production of various commodity chemicals (ethylene, isobutanol, sucrose), high-value chemicals (terpenoids), and biofuels (isobutene, 1-butanol, H 2 ) (Carbonell et al, 2019;Dienst et al, 2020;Durall et al, 2020;Liang et al, 2018;Liu et al, 2019;Miao et al, 2018;Mustila et al, 2021;Rodrigues & Lindberg, 2021;Thiel et al, 2018Thiel et al, , 2019. To this end, a minimal synthetic promoter for heterocyst specific expression of protein has been developed (Wegelius, Li, et al, 2018), and the possibilities and limitations for regulating target genes at translational level using selected Ribosome Binding Site (RBS) sequences have been evaluated (Thiel et al, 2018).…”

Section: Photosynthetic Cell Factoriesmentioning

confidence: 99%

NordAqua, a Nordic Center of Excellence to develop an algae‐based photosynthetic production platform

Allahverdiyeva

Aro

Bavel

et al. 2021

Physiologia Plantarum

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NordAqua is a multidisciplinary Nordic Center of Excellence funded by NordForsk Bioeconomy program (2017)(2018)(2019)(2020)(2021)(2022). The research center promotes Blue Bioeconomy and endeavours to reform the use of natural resources in a environmentally sustainable way. In this short communication, we summarize particular outcomes of the consortium. The key research progress of NordAqua includes (1) improving of photosynthetisis, (2) developing novel photosynthetic cell factories that function in a "solar-driven direct CO 2 capture to target bioproducts" mode, (3) promoting the diversity of Nordic cyanobacteria and algae as an abundant and resilient alternative for less sustainable forest biomass and for innovative production of biochemicals, and (4) improving the bio-based wastewater purification and nutrient recycling technologies to provide new tools for integrative circular economy platforms.A rapid replacement of carbon-intensive infrastructures with net-zero carbon alternatives is vital for mitigation of the climate crisis. Current bioeconomy approaches have been focused mainly on the replacement of fossil raw materials for bio-based commodities. To this end, Nordic countries have largely invested in wood biomass which, for a number of reasons, cannot be considered sustainable or sufficiently available to effectuate a major change. The NordAqua research center has taken a different approach and focuses on aquatic photosynthetic

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Isobutene production in Synechocystis sp. PCC 6803 by introducing α-ketoisocaproate dioxygenase from Rattus norvegicus

Cited by 24 publications

References 41 publications

Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production

Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production

Recent advances in metabolic engineering of microorganisms for advancing lignocellulose-derived biofuels

NordAqua, a Nordic Center of Excellence to develop an algae‐based photosynthetic production platform

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