Perspectives in Metabolic Engineering: Understanding Cellular Regulation Towards the Control of Metabolic Routes

Metabolic engineering seeks to redirect metabolic pathways through the modification of specific biochemical reactions or the introduction of new ones with the use of recombinant technology. Many of the chemicals synthesized via introduction of product-specific enzymes or the reconstruction of entire metabolic pathways into engineered hosts that can sustain production and can synthesize high yields of the desired product as yields of natural product-derived compounds are frequently low, and chemical processes can be both energy and material expensive; current endeavors have focused on using biologically derived processes as alternatives to chemical synthesis. Such economically favorable manufacturing processes pursue goals related to sustainable development and “green chemistry”. Metabolic engineering is a multidisciplinary approach, involving chemical engineering, molecular biology, biochemistry, and analytical chemistry. Recent advances in molecular biology, genome-scale models, theoretical understanding, and kinetic modeling has increased interest in using metabolic engineering to redirect metabolic fluxes for industrial and therapeutic purposes. The use of metabolic engineering has increased the productivity of industrially pertinent small molecules, alcohol-based biofuels, and biodiesel. Here, we highlight developments in the practical and theoretical strategies and technologies available for the metabolic engineering of simple systems and address current limitations.     

Development of a pre-concentrator-thermo-desorber/micro-gas chromatograph/mass spectrometer coupling for on-site analyses of emissions of volatile organic compounds from landfills

The development and use of a pre-concentrator-thermo-desorber/micro-gas chromatograph/mass spectrometer (μTD/μGC/MSD) coupling for the on-site analysis of VOCs in landfill gases are presented. The coupling has the advantage of analysing compounds with two detectors operated in series: the TCD (of the μGC) initially analyses the gas without destroying it, and then the MSD identifies the compounds. Due to the TCD response, the results were quantified with reference to toluene. The reliability of the analytical chain for quantitative analysis was validated by sampling two gaseous standards, including the EPA TO14 mixture, containing 39 compounds. With the OV1?μGC column, 24 compounds were identified and 16 correctly quantified. The repeatability of the measures estimated by their standard deviation was in the order of 1–2%. The detection limit was evaluated at 0.1?ppbv, for a 40?min pre-concentration on the Tenax of the μTD. The results of VOC analyses in the air of a landfill site obtained with the μTD/μGC/MSD coupling show its potential for on-site analyses: immediate results, high sensitivity, no storage for the samples, and measurements of pollution peaks.     

Triethylsilane as Hydride Donor in Ether Formation from Carbonyl Compounds

Triethylsilane is a convenient hydride source in the conversion of ketones or aldehydes to ethers under mild neutral conditions.     

A Study of the Molecular Weight Distribution of Poly(Amino Acid)s Synthesized by Diphenyl Phosphoryl Azide

Abstract

Direct polycondensations of β-benzyl-l-aspartate (Asp.Bz) and β-benzyl-l-glutamate (Glu.Bz) were carried out in the presence of diphenyl phosphoryl azide (DPPA) as a condensation agent and triethyl amine (TEA). Poly(amino acid)s were obtained by this convenient approach whose structure was confirmed by IR and ¹H-NMR spectroscopy. The effects of the monomer concentration, the polymerization time and temperature, the ratios [DPPA]/[monomer] and [TEA]/[monomer], and the solvent used on the molecular weight distribution of the polymer were studied. When the monomer concentrations were higher than 0.2 g/mL, poly(Asp.Bz) with a bimodal molecular weight distribution was obtained (M_w of 37,000 and M_w/M_n of 1.68). The polycondensations carried out in THF or in bulk provided the highest molecular weight (M_w ? 40,000). Several other amino acids were also polymerized by DPPA.     

Adsorption-desorption mechanism of phosphate by immobilized nano-sized magnetite layer: interface and bulk interactions

Phosphate adsorption mechanism by a homogenous porous layer of nano-sized magnetite particles immobilized onto granular activated carbon (nFe-GAC) was studied for both interface and bulk structures. X-ray Photoelectron Spectroscopy (XPS) analysis revealed phosphate bonding to the nFe-GAC predominantly through bidentate surface complexes. It was established that phosphate was adsorbed to the magnetite surface mainly via ligand exchange mechanism. Initially, phosphate was adsorbed by the active sites on the magnetite surface, after which it diffused into the interior of the nano-magnetite layer, as indicated by intraparticle diffusion model. This diffusion process continues regardless of interface interactions, revealing some of the outer magnetite binding sites for further phosphate uptake. Desorption, using NaOH solution, was found to be predominantly a surface reaction, at which hydroxyl ions replace the adsorbed phosphate ions only at the surface outer biding sites. Five successive fix-bed adsorption/regeneration cycles were successfully applied, without significant reduction in the nFe-GAC adsorption capacity and at high regeneration efficiency.     

Synthesis and photochemical properties of oligo-ortho-azobenzenes

Azobenzenes have attracted great interest in recent years because of their ability to change conformation upon irradiation. This property has been featured in several applications not only in organic chemistry but also in biology. Even though monoazobenzenes have been extensively studied and documented in the literature, only a few methods are available for the synthesis of oligo-ortho-azobenzenes. Also, their photochemical properties have not been reported so far. This study shows an efficient strategy for the preparation of oligo-ortho-azobenzenes and the investigation of their photochemical properties. It is demonstrated that the absorption spectra are highly influenced by the substituents. Interestingly, none of the ortho-bis-, tris-, or tetra-azobenzenes showed any E → Z isomerization. Only the ortho-nitrogen-substituted monoazobenzenes' photochromic behavior upon UV irradiation was observed.     

Chlorobis(pentafluoroethyl)phosphane: improved synthesis and molecular structure in the gas phase

(C₂F₅)₂PCl is now accessible through a significantly improved synthesis protocol starting from the technical product (C₂F₅)₃PF₂. (C₂F₅)₃PF₂ was reduced in the first step with NaBH₄ in a solvent‐free reaction at 120 °C. The product, P(C₂F₅)₃, was treated with an excess of an aqueous sodium hydroxide solution to afford the corresponding phosphinite salt Na⁺(C₂F₅)₂PO^? selectively under liberation of pentafluoroethane. Subsequent chlorination with PhPCl₄ resulted in the selective formation of (C₂F₅)₂PCl, which was isolated by fractional condensation in an overall yield of 66 %. The gas electron diffraction (GED) pattern for (C₂F₅)₂PCl was recorded and found to be described by a two‐conformer model. A quantum chemical investigation of the potential‐energy surface revealed the possible existence of many low‐energy conformers, each with a number of low‐frequency vibrational modes and therefore large‐amplitude motions. The conformer calculated to be most stable was also found to be most abundant by GED and comprised 61(5) % of the total. The molecular structure parameters determined by GED were in good agreement with those calculated at the MP2/TZVPP level of theory; the only significant difference was a discrepancy of about 3° in the C‐P‐C angle, which, for the lowest‐energy conformer, was refined to 98.2(4)° and was calculated to be 94.9°.     

Stress chain solutions in two-dimensional isostatic granular systems: fabric-dependent paths, leakage, and branching

The stress field equations for two-dimensional disordered isostatic granular materials are reformulated, giving new results beyond the commonly accepted force chains. Localized loads give rise to exactly determinable cones of influence, bounded by stress chains. Disorder couples same-source chains, attenuates stresses along chains, causes stress leakage from chains into the cone, and gives rise to branching. Chains from spatially separate sources do not interact. The formulation is convenient for computation, and several numerical solutions are presented.     

Ecologically friendly xanthan gum-PVA matrix for solid polymeric electrolytes

Two water-soluble and biodegradable polymers: xanthan gum (XG) and poly(vinyl alcohol) (PVA) were used to synthesize ecologically friendly solid polymer electrolyte (SPE) matrices. While XG is a natural polymer, PVA is a synthetic one, but both are colorless and form transparent membranes. To obtain ionic conductivity properties, the samples were doped with acetic acid and characterized by electrochemical impedance spectroscopy (EIS), X-ray diffraction, UV-Vis spectroscopy, and tensile test. The best results of ionic conductivity of 1.97 × 10^?4 and 7.41 × 10^?4 S/cm at room temperature and 80 °C, respectively, were obtained for the sample containing 55 wt% of acetic acid. Moreover, this electrolyte was found to be predominantly amorphous with transmittance in the visible region of 80% and absorbance values below 0.5 between 240 and 375 nm. Tensile test of this sample, applied up to 18 N of maximum force, resulted in strain of 2322% and Young’s modulus of 0.02 MPa. The obtained results showed that these new eco-friendly materials are promising for use as electrolytes in electrochemical devices.