Construction and analysis of a recombinant cyanobacterium expressing a chromosomally inserted gene for an ethylene-forming enzyme at the psbAI locus

Takahama, Kazutaka; Matsuoka, Masayoshi; Nagahama, Kazuhiro; Ogawa, Takahira

doi:10.1016/s1389-1723(03)80034-8

Cited by 112 publications

(68 citation statements)

References 13 publications

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“…Similarly, the Synechococcus PCC7942 strain harboring the Pseudomonas syringae gene ( efe ) encoding the ethylene-forming enzyme (Fukuda et al, 1992; Sakai et al, 1997), managed to introduce short nucleotide insertions in efe to stop ethylene production and recover a healthy growth (Takahama et al, 2003). Another recombinant Synechococcus PCC7942 strain could introduce a missense mutation in the E. coli atoD gene (acetoacetyl-CoA transferase) to decrease isopropanol production (Kusakabe et al, 2013).…”

Section: Resultsmentioning

confidence: 99%

Comparative Genomics of DNA Recombination and Repair in Cyanobacteria: Biotechnological Implications

2016

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Cyanobacteria are fascinating photosynthetic prokaryotes that are regarded as the ancestors of the plant chloroplast; the purveyors of oxygen and biomass for the food chain; and promising cell factories for an environmentally friendly production of chemicals. In colonizing most waters and soils of our planet, cyanobacteria are inevitably challenged by environmental stresses that generate DNA damages. Furthermore, many strains engineered for biotechnological purposes can use DNA recombination to stop synthesizing the biotechnological product. Hence, it is important to study DNA recombination and repair in cyanobacteria for both basic and applied research. This review reports what is known in a few widely studied model cyanobacteria and what can be inferred by mining the sequenced genomes of morphologically and physiologically diverse strains. We show that cyanobacteria possess many E. coli-like DNA recombination and repair genes, and possibly other genes not yet identified. E. coli-homolog genes are unevenly distributed in cyanobacteria, in agreement with their wide genome diversity. Many genes are extremely well conserved in cyanobacteria (mutMS, radA, recA, recFO, recG, recN, ruvABC, ssb, and uvrABCD), even in small genomes, suggesting that they encode the core DNA repair process. In addition to these core genes, the marine Prochlorococcus and Synechococcus strains harbor recBCD (DNA recombination), umuCD (mutational DNA replication), as well as the key SOS genes lexA (regulation of the SOS system) and sulA (postponing of cell division until completion of DNA reparation). Hence, these strains could possess an E. coli-type SOS system. In contrast, several cyanobacteria endowed with larger genomes lack typical SOS genes. For examples, the two studied Gloeobacter strains lack alkB, lexA, and sulA; and Synechococcus PCC7942 has neither lexA nor recCD. Furthermore, the Synechocystis PCC6803 lexA product does not regulate DNA repair genes. Collectively, these findings indicate that not all cyanobacteria have an E. coli-type SOS system. Also interestingly, several cyanobacteria possess multiple copies of E. coli-like DNA repair genes, such as Acaryochloris marina MBIC11017 (2 alkB, 3 ogt, 7 recA, 3 recD, 2 ssb, 3 umuC, 4 umuD, and 8 xerC), Cyanothece ATCC51142 (2 lexA and 4 ruvC), and Nostoc PCC7120 (2 ssb and 3 xerC).

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Section: Resultsmentioning

confidence: 99%

Comparative Genomics of DNA Recombination and Repair in Cyanobacteria: Biotechnological Implications

2016

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“…However, currently available tools for genetic modification allowed metabolic engineers to use cyanobacterium as a host for inserting exogenous genes for production of industrially important compounds. In fact, recent researches in cyanobacterial biological production have successfully achieved the production of targeted materials such as fatty acids [1], sugars [2], hydrogen [4], acetone [5], mannitol [6], ethylene [7,8], ethanol [9], isoprene [10], 3-hydroxybutyrate [11], 2,3-butanediol [12], isobutyraldehyde, isobutanol [13], isopropanol [14], 1-butanol [15], and 2-methy-1-butanol [16]. However, most of the bioprocess trials were not satisfactory for industrial application.…”

Section: Introductionmentioning

confidence: 99%

Molar-Based Targeted Metabolic Profiling of Cyanobacterial Strains with Potential for Biological Production

et al. 2014

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Recently, cyanobacteria have become one of the most attractive hosts for biochemical production due to its high proliferative ability and ease of genetic manipulation. Several researches aimed at biological production using modified cyanobacteria have been reported previously. However, to improve the yield of bioproducts, a thorough understanding of the intercellular metabolism of cyanobacteria is necessary. Metabolic profiling techniques have proven to be powerful tools for monitoring cellular metabolism of various organisms and can be applied to elucidate the details of cyanobacterial metabolism. In this study, we constructed a metabolic profiling method for cyanobacteria using 13C-labeled cell extracts as internal standards. Using this method, absolute concentrations of 84 metabolites were successfully determined in three cyanobacterial strains which are commonly used as background strains for metabolic engineering. By comparing the differences in basic metabolic potentials of the three cyanobacterial strains, we found a well-correlated relationship between intracellular energy state and growth in cyanobacteria. By integrating our results with the previously reported biological production pathways in cyanobacteria, we found putative limiting step of carbon flux. The information obtained from this study will not only help gain insights in cyanobacterial physiology but also serve as a foundation for future metabolic engineering studies using cyanobacteria.

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“…This approach can be applied to produce valuable chemicals, that the cyanobacteria host strains do not produce naturally, by constructing new biosynthetic pathways (6). Recently, cyanobacteria have been engineered to produce various chemicals, including isobutyraldehyde (14), isobutanol (14), 1-butanol (15), ethylene (16), isoprene (17), acetone (18), fatty acids (19), and fatty alcohols (20), through exogenous biosynthetic pathways. This approach in cyanobacteria is significantly less developed compared with a model organism such as Escherichia coli.…”

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confidence: 99%

Cyanobacterial conversion of carbon dioxide to 2,3-butanediol

Oliver

Machado

Yoneda

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

339

228

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Conversion of CO 2 for the synthesis of chemicals by photosynthetic organisms is an attractive target for establishing independence from fossil reserves. However, synthetic pathway construction in cyanobacteria is still in its infancy compared with model fermentative organisms. Here we systematically developed the 2,3-butanediol (23BD) biosynthetic pathway in Synechococcus elongatus PCC7942 as a model system to establish design methods for efficient exogenous chemical production in cyanobacteria. We identified 23BD as a target chemical with low host toxicity, and designed an oxygen-insensitive, cofactor-matched biosynthetic pathway coupled with irreversible enzymatic steps to create a driving force toward the target. Production of 23BD from CO 2 reached 2.38 g/L, which is a significant increase for chemical production from exogenous pathways in cyanobacteria. This work demonstrates that developing strong design methods can continue to increase chemical production in cyanobacteria.metabolic engineering | synthetic biology | biofuel | renewable energy A mid rising global energy demands and pressing environmental issues, interest is growing in the production of fuels and chemicals from renewable resources. Petroleum consumption reached 37.1 quadrillion BTU in the United States in 2008, of which a large majority (71%) was liquid fuel in the transportation sector. Petroleum and natural gas account for 99% of the feedstocks for chemicals, such as plastics, fertilizers, and pharmaceuticals in the chemical industry (1). Considering rapidly increasing world population and exhaustion of fossil fuels, the development of sustainable processes for energy and carbon capture to produce fuels and chemicals is crucial for human society.Energy and carbon capture by cyanobacteria is also directed toward mitigating increasing atmospheric CO 2 concentrations. According to the US Energy Information Administration (2), world energy-related CO 2 emissions in 2006 were 29 billion metric tons, which is an increase of 35% from 1990. Accelerating accumulation of atmospheric CO 2 is not only a result of increased emissions from world growth and intensifying carbon use, but also from a possible attenuation in the efficiency of the world's natural carbon sinks (3). As a result, atmospheric levels of CO 2 have increased by ∼25% over the past 150 y and it has become increasingly important to develop new technologies to reduce CO 2 emissions. Many creative solutions have been proposed and argued for carbon capture, each with varied environmental side-effects and costs (4). Sequestration by photosynthetic microorganisms in which CO 2 is biologically converted to valuable chemicals is an important addition to the toolbox for overall capture of CO 2 (5-7).Photosynthetic microorganisms, including cyanobacteria, are currently being engineered for platforms to convert solar energy to biochemicals renewably (5-7). These microorganisms possess many advantages over traditional terrestrial plants with regard to biochemical production. For example, the pho...

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Construction and analysis of a recombinant cyanobacterium expressing a chromosomally inserted gene for an ethylene-forming enzyme at the psbAI locus

Cited by 112 publications

References 13 publications

Comparative Genomics of DNA Recombination and Repair in Cyanobacteria: Biotechnological Implications

Comparative Genomics of DNA Recombination and Repair in Cyanobacteria: Biotechnological Implications

Molar-Based Targeted Metabolic Profiling of Cyanobacterial Strains with Potential for Biological Production

Cyanobacterial conversion of carbon dioxide to 2,3-butanediol

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