Efficient production of cyclic adenosine monophosphate from adenosine triphosphate by the N-terminal half of adenylate cyclase from Escherichia coli

In this study, we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate (cAMP) from adenosine triphosphate (ATP). First, adenylate cyclase from Escherichia coli MG1655 (EAC) and Bordetella Pertussis (BAC) were expressed in E. coli BL21(DE3) and comparatively analyzed for their activities. As a result, EAC from E. coli MG1655 exhibited a higher activity. However, amount of EAC were obtained in an insoluble form. Therefore, we expressed the first 446 amino acids of EAC (EAC446) to avoid the inclusion body. The effects of induction temperature, incubation time, and incubation pH were further evaluated to improve the expression of EAC446. Subsequently, the reaction process for the production of cAMP with ATP as a starting material was investigated. As none of cAMP was detected in the whole-cell based biocatalytic process, the reaction catalyzed by the crude enzyme was determined for cAMP production. What's more, the reaction temperature, reaction pH, metal ion additives and substrate concentration was optimized, and the maximum cAMP production of 18.45 g/L was achieved with a yield of 95.4% after bioconversion of 6 h.Graphical Abstract
Source: Chinese Journal of Chemical Engineering - Category: Chemistry Source Type: research

Related Links:

acute;a del Rayo Camacho-Corona Hechtia glomerata Zucc. is used both as a source of food and in ethnomedicine to treat various diseases derived from bacterial infections such as bronchitis, laryngitis, nephritis, whooping cough, urethritis, and sepsis. There are no previous reports about its chemistry and biological activities. Therefore, the aims of this study were to identify components from organic and aqueous extracts of H. glomerata and test the extracts and major isolate compounds against resistant bacteria. Hexane, CHCl3/MeOH, and aqueous extracts were prepared and analyzed by different chromatographic technique...
Source: Molecules - Category: Chemistry Authors: Tags: Article Source Type: research
Conclusions Current data indicates that Aa can colonize tooth surfaces above the gum-line and that this is an early event. In the section below we will describe how Aa can compete for nutritional elements with a multitude of rival bacteria in order to survive. Misconception 2: Nutritional Fastidious Nature of Aa Clinical Aa has been characterized as a fastidious microbe that requires CO2, serum, and specific carbohydrates such as glucose for its growth (34). It has been reported that Aa has a mandatory requirement for 5% CO2 for primary isolation, and that its growth is purported to be boosted by addition of serum (34)...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Publication date: Available online 20 February 2019Source: Phytochemistry LettersAuthor(s): Grażyna Łaska, Elwira Sieniawska, Łukasz Świątek, Jordan Zjawiony, Shabana Khan, Anastazja Boguszewska, Marcin Stocki, Mirosław Angielczyk, Małgorzata Polz-DacewiczAbstractPolemonium caeruleum L. is a Chinese medicinal plant used in the treatment of tuberculosis, whooping cough, fever, infertility, epilepsy, endometritis and insomnia. It was also used as sedative and anaesthetic agent. In this work the phytochemical investigation using LC-ESI-QTOF-MS and GC–MS techniques was done for chemical characterization of methano...
Source: Phytochemistry Letters - Category: Chemistry Source Type: research
This study has paved a way to understand the hydroxylation step of putrebactin synthesis which can be further investigated by studying its kinetic mechanism and physiological role.
Source: Protein Expression and Purification - Category: Biochemistry Source Type: research
Publication date: 5 September 2018Source: Journal of Hazardous Materials, Volume 357Author(s): Hua Fang, Lingxi Han, Houpu Zhang, Zhengnan Long, Lin Cai, Yunlong YuAbstractThe dissemination of antibiotic resistance genes (ARGs), human pathogenic bacteria (HPB), and antibiotic-resistant HPB (ARHPB) from animal feedlot to nearby environment poses a potentially high risk to environmental ecology and public health. Here, a metagenomic analysis was employed to explore the dissemination of ARGs, HPB, and ARHPB from a pig feedlot to surrounding stream and agricultural soils. In total, not detectable (ND)-1,628.4 μg/kg of ant...
Source: Journal of Hazardous Materials - Category: Environmental Health Source Type: research
Abstract Previously, we demonstrated that the ∼130-kDa CyaA-hemolysin (CyaA-Hly, Met482-Arg1706) from Bordetella pertussis was palmitoylated at Lys983 when co-expressed with CyaC-acyltransferase in Escherichia coli, and thus activated its hemolytic activity. Here, further investigation on a possible requirement of the N-terminal hydrophobic region (HP, Met482-Leu750) for toxin acylation was performed. The ∼100-kDa RTX (Repeat-in-ToXin) fragment (CyaA-RTX, Ala751-Arg1706) containing the K983-acylation region (AR, Ala751-Gln1000), but lacking HP, was co-produced with CyaC in E. coli. Hemolysis assay indicate...
Source: Biochemical and Biophysical Research communications - Category: Biochemistry Authors: Tags: Biochem Biophys Res Commun Source Type: research
Abstract Dynamic flux balance analysis (DFBA) has become an instrumental modeling tool for describing the dynamic behavior of bioprocesses. DFBA involves the maximization of a biologically meaningful objective subject to kinetic constraints on the rate of consumption/production of metabolites. In this paper, we propose a systematic data-based approach for finding both the biological objective function and a minimum set of active constraints necessary for matching the model predictions to the experimental data. The proposed algorithm accounts for the errors in the experiments and eliminates the need for ad hoc choi...
Source: Bioprocess and Biosystems Engineering - Category: Biomedical Engineering Authors: Tags: Bioprocess Biosyst Eng Source Type: research
zzi R Abstract Despite high vaccination coverage world-wide, whooping cough, a highly contagious disease caused by Bordetella pertussis, is recently increasing in occurrence suggesting that novel vaccine formulations targeted at the prevention of colonization and transmission should be investigated. In order to identify new candidates for inclusion in the acellular formulation, we used spontaneously released outer membrane vesicles (OMV) as a potential source of key adhesins. The enrichment of Bvg+ OMV with adhesins and the ability of anti-OMV serum to inhibit the adhesion of B. pertussis to lung epithelial cells ...
Source: Molecular and Cellular Proteomics : MCP - Category: Molecular Biology Authors: Tags: Mol Cell Proteomics Source Type: research
In conclusion, our results reveal the molecular mechanism underpinning activation of pertussis-like toxins, and we also identified differences in host target specificity.
Source: Journal of Biological Chemistry - Category: Chemistry Authors: Tags: Editors ' Picks Source Type: research
Conclusion This study revealed that the MPCE assay is a rapid, reliable, and high-throughput method with high specificity and sensitivity. This assay has great potential in the molecular epidemiological survey of respiratory pathogens.
Source: Biomedical and Environmental Sciences - Category: Biomedical Science Source Type: research
More News: Chemistry | China Health | Gastroenteritis | Study | Whooping Cough