A novel class of CoA-transferase involved in short-chain fatty acid metabolism in butyrate-producing human colonic bacteria

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Coenzyme A-transferase-independent butyrate re-assimilation in Clostridium acetobutylicum - evidence from a mathematical model

The hetero-dimeric CoA-transferase CtfA/B is believed to be crucial for the metabolic transition from acidogenesis to solventogenesis in Clostridium acetobutylicum as part of the industrial-relevant acetone-butanol-ethanol (ABE) fermentation. Here, the enzyme is assumed to mediate re-assimilation of acetate and butyrate during a pH-induced metabolic shift and to faciliate the first step of acetone formation from acetoacetyl-CoA. However, recent investigations using phosphate-limited continuous cultures have questioned this common dogma. To address the emerging experimental discrepancies, we investigated the mutant strain Cac-ctfA398s::CT using chemostat cultures. As a consequence of this mutation, the cells are unable to express functional ctfA and are thus lacking CoA-transferase activity. A mathematical model of the pH-induced metabolic shift, which was recently developed for the wild type, is used to analyse the observed behaviour of the mutant strain with a focus on re-assimilation activities for the two produced acids. Our theoretical analysis reveals that the ctfA mutant still re-assimilates butyrate, but not acetate. Based upon this finding, we conclude that C. acetobutylicum possesses a CoA-tranferase-independent butyrate uptake mechanism that is activated by decreasing pH levels. Furthermore, we observe that butanol formation is not inhibited under our experimental conditions, as suggested by previous batch culture experiments. In concordance with recent batch experiments, acetone formation is abolished in chemostat cultures using the ctfa mutant

Factors influencing citrus fruit scarring caused by Pezothrips kellyanus

[EN] Kelly s citrus thrips (KCT) Pezothrips kellyanus (Bagnall) (Thysanoptera: Thripidae) is a recently recorded cosmopolitan citrus pest, causing fruit scarring that results in downgrading of fruit. Due to the detrimental effects caused on fruits by KCT, we wanted to study some of the factors influencing fruit scarring. Specifically, the objectives were: (1) to determine the fruit development stage when citrus fruits are damaged by KCT and the population structure of KCT during this period, (2) to study the influence of temperature on intensity of damage, and finally, (3) to identify alternative host plants. KCT populations on flowers and fruitlets and alternate plant hosts were sampled in four citrus orchards from 2008 to 2010. The percentage of damaged fruits was also recorded. The exotic vine Araujia sericifera (Apocynaceae) was recorded as a new host for KCT. Thrips scarring started to increase at 350 650 degree-days (DD) above 10.2 C, coinciding with a peak abundance of the second instar larval stages over all 3 years of the study. The maximum percentage of larval stages of KCT was observed in the 3 years at about 500 DD, a period which corresponds to the end of May or early June. Variation in the severity of fruit scarring appeared to be related to air temperature. Temperature likely affects the synchronisation between the peak in abundance of KCT larvae, and the period when fruitlets are susceptible to thrips damage. Temperature can also influence the survival and development of KCT populations in citrus and other host plants in the citrus agro-ecosystem.The authors thank Alejandro Tena for his valuable suggestions and two anonymous referees for their careful review and helpful comments. We also extend our thanks to the owners of the commercial orchards for giving us permission to use their citrus orchards. The first author was awarded an FPI fellowship from the Polytechnic University of Valencia to obtain her PhD degree.Navarro Campos, C.; Pekas, A.; Aguilar Martí, MA.; Garcia Marí, F. (2013). Factors influencing citrus fruit scarring caused by Pezothrips kellyanus. Journal of Pest Science. (86):459-467. doi:10.1007/s10340-013-0489-7S45946786Baker GJ (2006) Kelly citrus thrips management. Fact sheet. Government of South Australia, primary industries and resources SA. http://www.sardi.sa.gov.au/__data/assets/pdf_file/0010/44875/kctfact_sheet.pdf . Accessed 16 July 2012Baker GJ, Jackman DJ, Keller M, MacGregor A, Purvis S (2002) Development of an integrated pest management system for thrips in Citrus. HAL Final Report CT97007. http://www.sardi.sa.gov.au/pestsdiseases/horticulture/horticultural_pests/kelly_citrus_thrips/research_report_1997-2000 . Accessed 16 July 2012Bedford ECG (1998) Thrips, wind and other blemishes. Citrus pests in the Republic of South Africa. 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Redox-switch regulatory mechanism of thiolase from Clostridium acetobutylicum

Thiolase is the first enzyme catalysing the condensation of two acetyl-coenzyme A (CoA) molecules to form acetoacetyl-CoA in a dedicated pathway towards the biosynthesis of n-butanol, an important solvent and biofuel. Here we elucidate the crystal structure of Clostridium acetobutylicum thiolase (CaTHL) in its reduced/oxidized states. CaTHL, unlike those from other aerobic bacteria such as Escherichia coli and Zoogloea ramegera, is regulated by the redox-switch modulation through reversible disulfide bond formation between two catalytic cysteine residues, Cys88 and Cys378. When CaTHL is overexpressed in wild-type C. acetobutylicum, butanol production is reduced due to the disturbance of acidogenic to solventogenic shift. The CaTHLV77Q/N153Y/A286K mutant, which is not able to form disulfide bonds, exhibits higher activity than wild-type CaTHL, and enhances butanol production upon overexpression. On the basis of these results, we suggest that CaTHL functions as a key enzyme in the regulation of the main metabolism of C. acetobutylicum through a redox-switch regulatory mechanism.close0

Comparative genomic and transcriptomic analysis revealed genetic characteristics related to solvent formation and xylose utilization in Clostridium acetobutylicum EA 2018

<p>Abstract</p> <p>Background</p> <p><it>Clostridium acetobutylicum</it>, a gram-positive and spore-forming anaerobe, is a major strain for the fermentative production of acetone, butanol and ethanol. But a previously isolated hyper-butanol producing strain <it>C. acetobutylicum </it>EA 2018 does not produce spores and has greater capability of solvent production, especially for butanol, than the type strain <it>C. acetobutylicum </it>ATCC 824.</p> <p>Results</p> <p>Complete genome of <it>C. acetobutylicum </it>EA 2018 was sequenced using Roche 454 pyrosequencing. Genomic comparison with ATCC 824 identified many variations which may contribute to the hyper-butanol producing characteristics in the EA 2018 strain, including a total of 46 deletion sites and 26 insertion sites. In addition, transcriptomic profiling of gene expression in EA 2018 relative to that of ATCC824 revealed expression-level changes of several key genes related to solvent formation. For example, <it>spo0A </it>and <it>adhEII </it>have higher expression level, and most of the acid formation related genes have lower expression level in EA 2018. Interestingly, the results also showed that the variation in CEA_G2622 (CAC2613 in ATCC 824), a putative transcriptional regulator involved in xylose utilization, might accelerate utilization of substrate xylose.</p> <p>Conclusions</p> <p>Comparative analysis of <it>C. acetobutylicum </it>hyper-butanol producing strain EA 2018 and type strain ATCC 824 at both genomic and transcriptomic levels, for the first time, provides molecular-level understanding of non-sporulation, higher solvent production and enhanced xylose utilization in the mutant EA 2018. The information could be valuable for further genetic modification of <it>C. acetobutylicum </it>for more effective butanol production.</p

Mathematical modelling of clostridial acetone-butanol-ethanol fermentation

Clostridial acetone-butanol-ethanol (ABE) fermentation features a remarkable shift in the cellular metabolic activity from acid formation, acidogenesis, to the production of industrial-relevant solvents, solventogensis. In recent decades, mathematical models have been employed to elucidate the complex interlinked regulation and conditions that determine these two distinct metabolic states and govern the transition between them. In this review, we discuss these models with a focus on the mechanisms controlling intra- and extracellular changes between acidogenesis and solventogenesis. In particular, we critically evaluate underlying model assumptions and predictions in the light of current experimental knowledge. Towards this end, we briefly introduce key ideas and assumptions applied in the discussed modelling approaches, but waive a comprehensive mathematical presentation. We distinguish between structural and dynamical models, which will be discussed in their chronological order to illustrate how new biological information facilitates the ‘evolution’ of mathematical models. Mathematical models and their analysis have significantly contributed to our knowledge of ABE fermentation and the underlying regulatory network which spans all levels of biological organization. However, the ties between the different levels of cellular regulation are not well understood. Furthermore, contradictory experimental and theoretical results challenge our current notion of ABE metabolic network structure. Thus, clostridial ABE fermentation still poses theoretical as well as experimental challenges which are best approached in close collaboration between modellers and experimentalists