Optimization of Tabersonine Methoxylation to Increase Vindoline Precursor Synthesis in Yeast Cell Factories

Plant specialized metabolites are widely used in the pharmaceutical industry, including the monoterpene indole alkaloids (MIAs) vinblastine and vincristine, which both display anticancer activity. Both compounds can be obtained through the chemical condensation of their precursors vindoline and catharanthine extracted from leaves of the Madagascar periwinkle. However, the extensive use of these molecules in chemotherapy increases precursor demand and results in recurrent shortages, explaining why the development of alternative production approaches, such microbial cell factories, is mandatory. In this context, the precursor-directed biosynthesis of vindoline from tabersonine in yeast-expressing heterologous biosynthetic genes is of particular interest but has not reached high production scales to date. To circumvent production bottlenecks, the metabolic flux was channeled towards the MIA of interest by modulating the copy number of the first two genes of the vindoline biosynthetic pathway, namely tabersonine 16-hydroxylase and tabersonine-16-O-methyltransferase. Increasing gene copies resulted in an optimized methoxylation of tabersonine and overcame the competition for tabersonine access with the third enzyme of the pathway, tabersonine 3-oxygenase, which exhibits a high substrate promiscuity. Through this approach, we successfully created a yeast strain that produces the fourth biosynthetic intermediate of vindoline without accumulation of other intermediates or undesired side-products. This optimization will probably pave the way towards the future development of yeast cell factories to produce vindoline at an industrial scale.

A New Method for the Synthesis of 3-Thiocyanatopyrazolo[1,5-a]pyrimidines

In this article, we demonstrate how an original effective “metal-free” and “chromatography-free” route for the synthesis of 3-thiocyanatopyrazolo[1,5-a]pyrimidines has been developed. It is based on electrooxidative (anodic) C–H thiocyanation of 5-aminopyrazoles by thiocyanate ion leading to 4-thiocyanato-5-aminopyrazoles (stage 1, yields up to 87%) following by their chemical condensation with 1,3-dicarbonyl compounds or their derivatives (stage 2, yields up to 96%). This method is equally effective for the synthesis of 3-thiocyanatopyrazolo[1,5-a]pyrimidines, both without substituents and with various donor (acceptor) substituents in the pyrimidine ring.

Influence of Magnetite Nanoparticles on Mechanical and Shielding Properties of Concrete

This paper presents an experimental study on the performance of shielding concrete with additives of magnetite nanoparticles. Two concretes with magnetite additives as well as one based concrete were tested. In order to achieve the high-performance concrete, all concrete mixes had a constant water/cement ratio of 0.45. In order to measure the mechanical properties, concrete samples were made in accordance with dimension such as 40 × 40 × 160 mm. But, for measurement of protective properties the concrete was made in accordance with dimension of rotary antennas such as 400 × 400 mm with a thickness of 10 mm. The nanoparticles Fe3O4 were synthesized by chemical condensation method. XRD have shown the presence of cubic structure of Fe3O4 spinel with crystallite size is equal to 130.0 Å. The TEM microphotograph shows that the Fe3O4 nanoparticles are spherical, the range of sizes is 12‒30 nm. The magnetic retardation suggests that the magnetite nanoparticles have superparamagnetic properties. This is explained by the fact that under the influence of external magnetic field, they are single-domain, in other words, they become uniformly magnetized throughout the volume. The additives of magnetite nanoparticles at a concentration of 0.5% mass have not a negative effect on flexural strength. The samples with additives of magnetite nanoparticles showed better shielding of microwave radiation in the frequency range from 0.7 GHz to 13 GHz. The maximum efficiency of suppression of electromagnetic disturbance is equal to 19.9 dB at a frequency of 1.5 GHz with a thickness of 10 mm.

Controlled Stepwise Growth of Siloxane Chains Using Bivalent Building Units With Different Functionalities

Organic/inorganic hybrids of silicon and their subsequent chemical modification are of interest for tailoring and structuring surfaces on the nanoscale. The formation of monolayers on hydroxylated silicon surfaces was employed to synthesize molecular dimethylsiloxane chains by wet-chemical condensation reactions, using dimethylmonochlorosilane as the precursor. The SiH group of the resulting dimethylsilyl termination could be selectively oxidized to the SiOH group, which opened the possibility of bonding another species. By repeating the condensation and oxidation cycle the stepwise growth of one-dimensional dimethylsiloxane chains was achieved. The ongoing chain growth was characterized by attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy, x-ray photoelectron spectroscopy (XPS), spectroscopic ellipsometry (SE), and determination of the surface energy by contact-angle experiments.