Isoprene production by Rhodobacter sphaeroides and its antimicrobial activity

Yu, Jaeyoung; Moon, Se-Kwon; Kim, Yang‐Hoon; Min, Jiho

doi:10.1016/j.resmic.2022.103938

Cited by 6 publications

(6 citation statements)

References 49 publications

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“…The methods from primary fixation to ethanol dehydration were performed according to a previously published protocol ( 32 ). The dehydrated samples were transitioned twice with 100% propylene oxide for 5 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

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Increase CO ₂ recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Yu,

Shin,

Kim

et al. 2023

Microbiol Spectr

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One of the current promising solutions to address problems of CO 2 emissions is the development of biological carbon fixation strategies. This strategy leads to higher chemical biosynthetic productivity through improved biological CO 2 fixation. In a previous study, we introduced the Calvin-Benson Bassham (CBB) genes of the photosynthetic bacterium Cereibacter sphaeroides into Escherichia coli to develop a strain capable of endogenous CO 2 recycling by heterologous expression of the CBB genes. However, the heterologous expression of recombinant proteins in E. coli is often hampered by inclusion bodies (IB), which are protein aggregates. Various factors contribute to IB formation, including host cell metabolism, protein synthesis, transformation machinery, and target protein properties. In this study, we investigated the influence of environmental conditions, particularly culture temperature, on IB formation and CO 2 recycling in the CBB strain. As a result, by reducing the culture temperature from 37°C to 30°C, a significant suppression of IB formation was achieved, resulting in a remarkable decrease in CO 2 release by approximately 5.76 times. In addition, interestingly, an enhancement in the accumulation of pyruvate by approximately 2.3 times was observed at the same time. These results demonstrate the simultaneous improvements in CO 2 recycling and the synthesis of organic acids achieved through temperature control. Based on the findings of this study, we believe that temperature control would be a promising approach to increasing chemical biosynthetic productivity as well as biological CO 2 fixation activity in integrated autotrophic biorefinery strategies. IMPORTANCE In a previous study, we successfully engineered Escherichia coli capable of endogenous CO 2 recycling through the heterologous expression of the Calvin-Benson Bassham genes. Establishing an efficient gene expression environment for recombinant strains is crucial, on par with the importance of metabolic engineering design. Therefore, the primary objective of this study was to further mitigate greenhouse gas emissions by investigating the effects of culture temperature on the formation of inclusion bodies (IB) and CO 2 fixation activity in the engineered bacterial strain. The findings demonstrate that lowering the culture temperature effectively suppresses IB formation, enhances CO 2 recycling, and concurrently increases the accumulation of organic acids. This temperature control approach, without adding or modifying compounds, is both convenient and efficient for enhancing CO 2 recycling. As such, additional optimization of various environmental parameters holds promise for further enhancing the performance of recombinant strains efficiently.

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Section: Methodsmentioning

confidence: 99%

“…According to a previously published protocol, LC/MS/MS analysis was performed ( 32 ). Pyruvate standard was purchased from Sigma-Aldrich (St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

Increase CO ₂ recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Yu,

Shin,

Kim

et al. 2023

Microbiol Spectr

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“…This could be tested further by challenging these homoacetogenic microcosms with other alkenes and dialkenes, as they may reduce more than just isoprene. Soil‐dwelling and aquatic bacteria also produce isoprene by the methylerythritol phosphate (MEP) pathway (Wagner et al, 2000; Yu et al, 2022), permitting isoprene formation in the absence of sunlight, potentially supplying isoprene to soil‐dwelling anaerobes closer to the oxic‐anoxic interface. Rhodobacter sphaeroides , a purple non‐sulfur bacterium, also produced isoprene via the MEP pathway, with the authors noting an antimicrobial effect of isoprene on Gram‐positive and Gram‐negative bacteria (Yu et al, 2022).…”

Section: Ecology Of Isoprene‐degrading Bacteria: An Untapped Metaboli...mentioning

confidence: 99%

“…Soil‐dwelling and aquatic bacteria also produce isoprene by the methylerythritol phosphate (MEP) pathway (Wagner et al, 2000; Yu et al, 2022), permitting isoprene formation in the absence of sunlight, potentially supplying isoprene to soil‐dwelling anaerobes closer to the oxic‐anoxic interface. Rhodobacter sphaeroides , a purple non‐sulfur bacterium, also produced isoprene via the MEP pathway, with the authors noting an antimicrobial effect of isoprene on Gram‐positive and Gram‐negative bacteria (Yu et al, 2022). This raises further questions as to the role of isoprene emission in bacteria, and also to the benefits of isoprene consumption by bacteria.…”

Section: Ecology Of Isoprene‐degrading Bacteria: An Untapped Metaboli...mentioning

confidence: 99%

Peering down the sink: A review of isoprene metabolism by bacteria

Dawson

Crombie

Jansen

et al. 2023

Environmental Microbiology

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Isoprene (2-methyl-1,3-butadiene) is emitted to the atmosphere each year in sufficient quantities to rival methane (>500 Tg C yr À1 ), primarily due to emission by trees and other plants. Chemical reactions of isoprene with other atmospheric compounds, such as hydroxyl radicals and inorganic nitrogen species (NO x ), have implications for global warming and local air quality, respectively. For many years, it has been estimated that soil-dwelling bacteria consume a significant amount of isoprene ($20 Tg C yr À1 ), but the mechanisms underlying the biological sink for isoprene have been poorly understood. Studies have indicated or confirmed the ability of diverse bacterial genera to degrade isoprene, whether by the canonical iso-type isoprene degradation pathway or through other less wellcharacterized mechanisms. Here, we review current knowledge of isoprene metabolism and highlight key areas for further research. In particular, examples of isoprene-degraders that do not utilize the isoprene monooxygenase have been identified in recent years. This has fascinating implications both for the mechanism of isoprene uptake by bacteria, and also for the ecology of isoprene-degraders in the environments.

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“…One of the microorganisms that can synthesize CoQ 10 efficiently and has been used in industrial production is Rhodobacter sphaeroides [ 13 , 14 ]. It is a gram-negative purple non sulfur bacterium (PNSB) that can grow aerobically, anaerobically, photoautotrophically, and heterotrophically [ 15 – 17 ]. It has many other functions, such as biological hydrogen production, synthesis of carotenoids, fixation of CO 2 and N 2 , remediation of heavy metal pollution and power generation [ 18 – 21 ].…”

Section: Introductionmentioning

confidence: 99%

Metabolic flux analysis of coenzyme Q10 synthesized by Rhodobacter sphaeroides under the influence of different pH regulators

Xiao,

Zheng,

Zhou

et al. 2023

Microb Cell Fact

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Coenzyme Q10 (CoQ10) is crucial for human beings, especially in the fields of biology and medicine. The aim of this experiment was to investigate the conditions for increasing CoQ10 production. At present, microbial fermentation is the main production method of CoQ10, and the production process of microbial CoQ10 metabolism control fermentation is very critical. Metabolic flux is one of the most important determinants of cell physiology in metabolic engineering. Metabolic flux analysis (MFA) is used to estimate the intracellular flux in metabolic networks. In this experiment, Rhodobacter sphaeroides was used as the research object to analyze the effects of aqueous ammonia (NH3·H2O) and calcium carbonate (CaCO3) on the metabolic flux of CoQ10. When CaCO3 was used to adjust the pH, the yield of CoQ10 was 274.43 mg·L−1 (8.71 mg·g−1 DCW), which was higher than that of NH3·H2O adjustment. The results indicated that when CaCO3 was used to adjust pH, more glucose-6-phosphate (G6P) entered the pentose phosphate (HMP) pathway and produced more NADPH, which enhanced the synthesis of CoQ10. At the chorismic acid node, more metabolic fluxes were involved in the synthesis of p-hydroxybenzoic acid (pHBA; the synthetic precursor of CoQ10), enhancing the anabolic flow of CoQ10. In addition, Ca2+ produced by the reaction of CaCO3 with organic acids promotes the synthesis of CoQ10. In summary, the use of CaCO3 adjustment is more favorable for the synthesis of CoQ10 by R. sphaeroides than NH3·H2O adjustment. The migration of metabolic flux caused by the perturbation of culture conditions was analyzed to compare the changes in the distribution of intracellular metabolic fluxes for the synthesis of CoQ10. Thus, the main nodes of the metabolic network were identified as G6P and chorismic acid. This provides a theoretical basis for the modification of genes related to the CoQ10 synthesis pathway.

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Isoprene production by Rhodobacter sphaeroides and its antimicrobial activity

Cited by 6 publications

References 49 publications

Increase CO ₂ recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Increase CO ₂ recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Peering down the sink: A review of isoprene metabolism by bacteria

Metabolic flux analysis of coenzyme Q10 synthesized by Rhodobacter sphaeroides under the influence of different pH regulators

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Isoprene production by Rhodobacter sphaeroides and its antimicrobial activity

Cited by 6 publications

References 49 publications

Increase CO 2 recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Increase CO 2 recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Peering down the sink: A review of isoprene metabolism by bacteria

Metabolic flux analysis of coenzyme Q10 synthesized by Rhodobacter sphaeroides under the influence of different pH regulators

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Product

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Increase CO ₂ recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction

Increase CO ₂ recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction