Directed Evolution Empowered Redesign of Natural Proteins for the Sustainable Production of Chemicals and Pharmaceuticals

Directed evolution has advanced into a standard industrial “tool” to tailor naturally occurring proteins for a variety of biotechnological applications, thus enabling product valorization and bringing societal benefits across industrial sectors. Examples are sustainable enzymatic production processes for chemicals, pharmaceuticals, or applications in the food, feed, and laundry industries. In essence, directed evolution has contributed to sustainable industrial processes that fuel the transition from a fossil‐based economy to a biobased economy utilizing renewable resources. Phage display technologies represent a comparable breakthrough that allow for the directed evolution of binding proteins by physical coupling between a phenotype and the respective genotype, thus enabling the identification of highly selective antibodies for a broad variety of applications in diagnostics and therapy.     

Nanosecond surface interferometry measurements on designed and commercial polymers

The effect of the ablation mechanism on surface morphology changes during an ablation process was studied by comparing three different polymers: a triazene polymer, a polyimide and poly(methylmethacrylate) (PMMA) with nanosecond surface interferometry. The triazene polymer, for which only indications for a photochemical ablation mechanism had been detected in previous studies, revealed no surface swelling, which could be attributed to a thermal ablation mechanism. For polyimide, a photothermal ablation mechanism is usually used to describe the ablation process at irradiation wavelengths 248 nm. However, the interferometric measurements do not show any surface swelling, which would be a clear indication for a thermal ablation mechanism. A surface swelling was only detected for PMMA with irradiation at 248 nm and fluences below the threshold of permanent surface modification. The detected phase shift, which is proportional to the change of the film thickness and the refractive index, can be explained by the opposite signs of the thermal expansion coefficient and the thermal refractive-index coefficient. PACS 52.38.Mf; 42.87.Bg; 71.20.Rv     

High-throughput assay of (<Emphasis Type="Italic">R</Emphasis>)-phenylacetylcarbinol synthesized by pyruvate decarboxylase

A novel assay has been developed for the detection of ( R)-phenylacetylcarbinol, ( R)-PAC, a chiral intermediate in the industrial synthesis of ephedrine. It is the product of a biotransformation of benzaldehyde catalysed by the enzyme pyruvate decarboxylase. The assay, using 2,3,5-triphenyltetrazolium chloride, enables high-throughput photometric analysis of the activity of the enzyme thus avoiding time-consuming chromatographic procedures.     

Biocatalytic racemization of aliphatic, arylaliphatic, and aromatic alpha-hydroxycarboxylic acids

[reaction: see text] Biocatalytic racemization of a range of aliphatic, (aryl)aliphatic, and aromatic alpha-hydroxycarboxylic acids was accomplished by using whole resting cells of a range of Lactobacillus spp. The mild (physiological) reaction conditions ensured an essentially "clean" isomerization in the absence of side reactions, such as elimination or decomposition. Whereas straight-chain aliphatic 2-hydroxycarboxylic acids were racemized with excellent rates (up to 85% relative to lactate), steric hindrance was observed for branched-chain analogues. Good rates were observed for aryl-alkyl derivatives, such as 3-phenyllactic acid (up to 59%) and 4-phenyl-2-hydroxybutanoic acid (up to 47%). In addition, also mandelate and its o-chloro analogue were accepted at a fair rate (45%). This biocatalytic racemization represents an important tool for the deracemization of a number of pharmaceutically important building blocks.     

Evaluation of adsorbent materials for heat pump and thermal energy storage applications in open systems

The evaluation of solid adsorbents in open sorption systems for heating, cooling and thermal energy storage (TES) applications is crucial for the ecological and economical performance of these systems. An appropriate adsorbent has to reach the temperature limit given by the heating/cooling system of the consumer. It has to provide high energy efficiency and a high energy density for storage applications. A method for an easy evaluation of different adsorbents for a specific application has been developed. The method is based on the adsorption equilibrium of the adsorbent and water vapor. The crucial property for the discussed field of applications is the differential heat of adsorption. Criteria for the evaluation of the adsorbent are the breakthrough curves (responsible for the dynamics of the process), the possible temperature lift (or the dehumidification) of the air, the thermal COP and the storage capacity.     

The central role of the metal ion for photoactivity: Zn– vs. Ni–Mabiq

Photoredox catalysts are integral components of artificial photosystems, and have recently emerged as powerful tools for catalysing numerous organic reactions. However, the development of inexpensive and efficient earth-abundant photoredox catalysts remains a challenge. We here present the photochemical and photophysical properties of a Ni–Mabiq catalyst ([Ni^II(Mabiq)]OTf (1); Mabiq = 2-4:6-8-bis(3,3,4,4-tetramethyldihydropyrrolo)-10-15-(2,2-biquinazolino)-[15]-1,3,5,8,10,14-hexaene1,3,7,9,11,14-N₆)—and of a Zn-containing analogue ([Zn^II(Mabiq)OTf] (2))—using steady state and time resolved optical spectroscopy, time-dependent density functional theory (TDDFT) calculations, and reactivity studies. The Ni and Zn complexes exhibit similar absorption spectra, but markedly different photochemical properties. These differences arise because the excited states of 2 are ligand-localized, whereas metal-centered states account for the photoactivity of 1. The distinct properties of the Ni and Zn complexes are manifest in their behavior in the photo-driven aza-Henry reaction and oxidative coupling of methoxybenzylamine.

The development of earth-abundant photoredox catalysts remains a challenge. Studies of Ni- and Zn-Mabiq complexes demonstrate how the coordinating metal ion influences the photochemistry, photodynamics and reactivity of photocatalysts.