Insights into the stability and substrate specificity of the E. coli aerobic β-oxidation trifunctional enzyme complex

Sah‐Teli, Shiv K.; Hynönen, Mikko J.; Sulu, R.; Dalwani, Subhadra; Schmitz, W.; Wierenga, Rik K.; Radha, Venkatesan

doi:10.1016/j.jsb.2020.107494

Cited by 13 publications

(15 citation statements)

References 55 publications

(64 reference statements)

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“…We are only aware of one other case of a Zn‐finger protein involved in a catalytic process, namely in adenylate kinase from Bacillus stearothermophilus (https://doi.org/10.2210/pdb1ZIN/pdb) where it provides a structural lid for the active site [35]. Another example where protein assembly is required to attain thiolase activity was reported for a trifunctional β‐oxidation enzyme of E. coli which depends on forming a complex with a subunit contributing enoyl‐CoA hydratase and alcohol dehydrogenase functionalities [36].…”

Section: Discussionmentioning

confidence: 99%

Finis tolueni: a new type of thiolase with an integrated Zn‐finger subunit catalyzes the final step of anaerobic toluene metabolism

et al. 2022

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Anaerobic toluene degradation involves β‐oxidation of the first intermediate (R)‐2‐benzylsuccinate to succinyl‐CoA and benzoyl‐CoA. Here, we characterize the last enzyme of this pathway, (S)‐2‐benzoylsuccinyl‐CoA thiolase (BbsAB). Although benzoylsuccinyl‐CoA is not available for enzyme assays, the recombinantly produced enzymes from two different species showed the reverse activity, benzoylsuccinyl‐CoA formation from benzoyl‐CoA and succinyl‐CoA. Activity depended on the presence of both subunits, the thiolase family member BbsB and the Zn‐finger protein BbsA, which is affiliated to the DUF35 family of unknown function. We determined the structure of BbsAB from Geobacter metallireducens with and without bound CoA at 1.7 and 2.0 Å resolution, respectively. CoA binding into the well‐known thiolase cavity triggers an induced‐fit movement of the highly disordered covering loop, resulting in its rigidification by forming multiple interactions to the outstretched CoA moiety. This event is coupled with an 8 Å movement of an adjacent hairpin loop of BbsB and the C‐terminal domain of BbsA. Thereby, CoA is placed into a catalytically productive conformation, and a putative second CoA binding site involving BbsA and the partner BbsB′ subunit is simultaneously formed that also reaches the active center. Therefore, while maintaining the standard thioester‐dependent Claisen‐type mechanism, BbsAB represents a new type of thiolase.

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Section: Discussionmentioning

confidence: 99%

Finis tolueni: a new type of thiolase with an integrated Zn‐finger subunit catalyzes the final step of anaerobic toluene metabolism

et al. 2022

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“…Three-dimensional structures have been obtained for bacterial and human TFE ( Figure 4 ) [ 23 , 24 , 25 ]. In addition, the structure of FadB from E. coli was published recently [ 26 ]. The FadBA complex is a hetero-tetramer composed of two copies of the 78 kDa multifunctional enzyme FadB, and two copies of the 42 kDa 3-ketoacyl-CoA thiolase FadA.…”

Section: Pathways Of Fatty Acid Degradationmentioning

confidence: 99%

“…Despite their homology, in vitro analysis suggested that FadIJ and FadBA complexes present many different features [ 26 , 59 ]. FadIJ was modeled as a hetero-octameric enzyme, possibly membrane-bound.…”

Section: Pathways Of Fatty Acid Degradationmentioning

confidence: 99%

Degradation of Exogenous Fatty Acids in Escherichia coli

2022

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Many bacteria possess all the machineries required to grow on fatty acids (FA) as a unique source of carbon and energy. FA degradation proceeds through the ß-oxidation cycle that produces acetyl-CoA and reduced NADH and FADH cofactors. In addition to all the enzymes required for ß-oxidation, FA degradation also depends on sophisticated systems for its genetic regulation and for FA transport. The fact that these machineries are conserved in bacteria suggests a crucial role in environmental conditions, especially for enterobacteria. Bacteria also possess specific enzymes required for the degradation of FAs from their environment, again showing the importance of this metabolism for bacterial adaptation. In this review, we mainly describe FA degradation in the Escherichia coli model, and along the way, we highlight and discuss important aspects of this metabolism that are still unclear. We do not detail exhaustively the diversity of the machineries found in other bacteria, but we mention them if they bring additional information or enlightenment on specific aspects.

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“…Generally, the level of ATP is directly proportional to the intensity of the emitted light. By monitoring the level of ATP, the physiological and pathological properties of cells can be obtained [ 20 , 21 ]. In the area of bacteria detection, the ATP luminescent method can be used to measure the amount of bacteria conveniently and quickly.…”

Section: Introductionmentioning

confidence: 99%

Microfluidics for the rapid detection of Escherichia coli O157:H7 using antibody-coated microspheres

Shopsin

Sun

et al. 2021

Bioengineered

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This study developed a novel method for the rapid detection of Escherichia coli (E. coli) O157:H7 on a microfluidic platform. First, the concentration of bacteria in a sample was determined with the adenosine triphosphate (ATP) method. Then, the specific detection of E. coli was achieved in a microfluidic chip by the immune-microsphere technique. The influences of the culture time, flow rate and capture time on the detection of the target bacteria were investigated systematically. Generally, with increasing capture time, more bacteria could be captured by the microspheres, which had a positive effect on bacterial detection. Furthermore, the sensitivity and specificity of the method were also tested. The results showed that this method could specifically detect E. coli with a sensitivity as high as 49.1 cfu/μL; the consumption of bacteria was 1 μL, and the reagent was at the microliter level. The testing time can be controlled within one and a half hours, and the cost of testing was approximately RMB 10. The method described in this article is simple and accurate and has great application value in bacterial detection for medical diagnostics.

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Insights into the stability and substrate specificity of the E. coli aerobic β-oxidation trifunctional enzyme complex

Cited by 13 publications

References 55 publications

Finis tolueni: a new type of thiolase with an integrated Zn‐finger subunit catalyzes the final step of anaerobic toluene metabolism

Finis tolueni: a new type of thiolase with an integrated Zn‐finger subunit catalyzes the final step of anaerobic toluene metabolism

Degradation of Exogenous Fatty Acids in Escherichia coli

Microfluidics for the rapid detection of Escherichia coli O157:H7 using antibody-coated microspheres

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