Artificial Splitting of a Non‐Ribosomal Peptide Synthetase by Inserting Natural Docking Domains

The interaction in multisubunit non‐ribosomal peptide synthetases (NRPSs) is mediated by docking domains that ensure the correct subunit‐to‐subunit interaction. We introduced natural docking domains into the three‐module xefoampeptide synthetase (XfpS) to create two to three artificial NRPS XfpS subunits. The enzymatic performance of the split biosynthesis was measured by absolute quantification of the products by HPLC‐ESI‐MS. The connecting role of the docking domains was probed by deleting integral parts of them. The peptide production data was compared to soluble protein amounts of the NRPS using SDS‐PAGE. Reduced peptide synthesis was not a result of reduced soluble NRPS concentration but a consequence of the deletion of vital docking domain parts. Splitting the xefoampeptide biosynthesis polypeptide by introducing docking domains was feasible and resulted in higher amounts of product in one of the two tested split‐module cases compared to the full‐length wild‐type enzyme.     

吴茱萸次碱的合成进展

系统地总结了吴茱萸次碱的合成方法. 根据构建环的种类不同, 把合成方法分为七类, 大部分方法都是以色胺和邻氨基苯甲酸或其衍生物为起始原料, 步骤较长, 适合工业生产的简便、经济适用的方法还有待于进一步研究探索.     

Synthesis and activity of two trifluorinated analogues of 1-deoxy-d-xylulose 5-phosphate

An improved synthesis of 1,1,1-trifluoro-1-deoxy-d-xylulose 5-phosphate and an access to the reduced diastereomer mixture analogues 1,1,1-trifluoro-1-deoxy-d-xylitol 5-phosphate and 1,1,1-trifluoro-1-deoxy-d-lyxitol 5-phosphate are described. Inhibitor activity of all compounds on the MEP pathway for isoprenoid biosynthesis was evaluated.     

A highly efficient synthesis of [1-C, O]- and [1-C, H2]-glycerol for the elucidation of biosynthetic pathways

Labeled glycerol is a widely used biochemical probe to investigate biosynthetic pathways. A highly efficient synthesis of [1-¹³C, ¹⁸O]- and [1-¹³C, ²H₂]-glycerol is described in which the ¹³C label is introduced using cyanide. The ¹⁸O label was introduced by a Pinner synthesis and reduction of the ester 5 allowed incorporation of the ²H labels.     

High Hydrostatic Pressure to Increase the Biosynthesis and Extraction of Phenolic Compounds in Food: A Review

Phenolic compounds from fruits and vegetables have shown antioxidant, anticancer, anti-inflammatory, among other beneficial properties for human health. All these benefits have motivated multiple studies about preserving, extracting, and even increasing the concentration of these compounds in foods. A diverse group of vegetable products treated with High Hydrostatic Pressure (HHP) at different pressure and time have shown higher phenolic content than their untreated counterparts. The increments have been associated with an improvement in their extraction from cellular tissues and even with the activation of the biosynthetic pathway for their production. The application of HHP from 500 to 600 MPa, has been shown to cause cell wall disruption facilitating the release of phenolic compounds from cell compartments. HPP treatments ranging from 15 to 100 MPa during 10–20 min at room temperature have produced changes in phenolic biosynthesis with increments up to 155%. This review analyzes the use of HHP as a method to increase the phenolic content in vegetable systems. Phenolic content changes are associated with either an immediate stress response, with a consequent improvement in their extraction from cellular tissues, or a late stress response that activates the biosynthetic pathways of phenolics in plants.     

Extracellular biosynthesis of monodispersed gold nanoparticles by a SAM capping route

Monodispersed gold nanoparticles capped with a self-assembled monolayer of dodecanethiol were biosynthesized extracellularly by an efficient, simple, and environmental friendly procedure, which involved the use of Bacillus megatherium D01 as the reducing agent and the use of dodecanethiol as the capping ligand at 26 °C. The kinetics of gold nanoparticle formation was followed by transmission electron microscope (TEM) and UV-vis spectroscopy. It was shown that reaction time was an important parameter in controlling the morphology of gold nanoparticles. The effect of thiol on the shape, size, and dispersity of gold nanoparticles was also studied. The results showed that the presence of thiol during the biosynthesis could induce the formation of small size gold nanoparticles (<2.5 nm), hold the shape of spherical nanoparticles, and promote the monodispersity of nanoparticles. Through the modulation of reaction time and the use of thiol, monodispersed spherical gold nanoparticles capped with thiol of 1.9 ± 0.8 nm size were formed by using Bacillus megatherium D01.     

AibA/AibB Induces an Intramolecular Decarboxylation in Isovalerate Biosynthesis by Myxococcus xanthus

Isovaleryl coenzyme A (IV‐CoA) is an important precursor for iso ‐fatty acids and lipids. It acts in the development of myxobacteria, which can produce this compound from acetyl‐CoA through alternative IV‐CoA biosynthesis (aib). A central reaction of aib is catalyzed by AibA/AibB, which acts as a cofactor‐free decarboxylase despite belonging to the family of CoA‐transferases. We developed an efficient expression system for AibA/AibB that allowed the determination of high‐resolution crystal structures in complex with different ligands. Through mutational studies, we show that an active‐site cysteine previously proposed to be involved in decarboxylation is not required for activity. Instead, AibA/AibB seems to induce an intramolecular decarboxylation by binding its substrate in a hydrophobic cavity and forcing it into a bent conformation. Our study opens opportunities for synthetic biology studies, since AibA/AibB may be suitable for the production of isobutene, a precursor of biofuels and chemicals.