Environmental concerns and bioaccumulation of psychiatric drugs in water bodies - Conventional versus biocatalytic systems of mitigation.

The COVID-19 pandemic has brought increments in market sales and prescription of medicines commonly used to treat mental health disorders, such as depression, anxiety, stress, and related problems. The increasing use of these drugs, named psychiatric drugs, has led to their persistence in aquatic systems (bioaccumulation), since they are recalcitrant to conventional physical and chemical treatments typically used in wastewater treatment plants. An emerging environmental concern caused by the bioaccumulation of psychiatric drugs has been attributed to the potential ecological and toxicological risk that these medicines might have over human health, animals, and plants. Thus, by the application of biocatalysis-assisted techniques, it is possible to efficiently remove psychiatric drugs from water. Biocatalysis, is a widely employed and highly efficient process implemented in the biotransformation of a wide range of contaminants, since it has important differences in terms of catalytic behavior, compared to common treatment techniques, including photodegradation, Fenton, and thermal treatments, among others. Moreover, it is noticed the importance to monitor transformation products of degradation and biodegradation, since according to the applied removal technique, different toxic transformation products have been reported to appear after the application of physical and chemical procedures. In addition, this work deals with the discussion of differences existing between high- and low-income countries, according to their environmental regulations regarding waste management policies, especially waste of the drug industry.

New Additions to the Arsenal of Biocatalysts for Noncanonical Amino Acid Synthesis.

Noncanonical amino acids (ncAAs) merge the conformational behavior and native interactions of proteinogenic amino acids with nonnative chemical motifs and have proven invaluable in developing modern therapeutics. This blending of native and nonnative characteristics has resulted in essential drugs like nirmatrelvir, which comprises three ncAAs and is used to treat COVID-19. Enzymes are appearing prominently in recent syntheses of ncAAs, where they demonstrate impressive control over the stereocenters and functional groups found therein. Here we review recent efforts to expand the biocatalyst arsenal for synthesizing ncAAs with natural enzymes. We also discuss how new-to-nature enzymes can contribute to this effort by catalyzing reactions inspired by the vast repertoire of chemical catalysis and acting on substrates that would otherwise not be used in synthesizing ncAAs. Abiotic enzyme-catalyzed reactions exploit the selectivity afforded by a macromolecular catalyst to access molecules not available to natural enzymes and perhaps not even chemical catalysis.

Mechanism of reaction of RNA-dependent RNA polymerase from SARS-CoV-2.

We combine molecular dynamics, statistical mechanics, and hybrid quantum mechanics/molecular mechanics simulations to describe mechanistically the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA-dependent RNA polymerase (RdRp). Our study analyzes the binding mode of both natural triphosphate substrates as well as remdesivir triphosphate (the active form of drug), which is bound preferentially over ATP by RdRp while being poorly recognized by human RNA polymerase II (RNA Pol II). A comparison of incorporation rates between natural and antiviral nucleotides shows that remdesivir is incorporated more slowly into the nascent RNA compared with ATP, leading to an RNA duplex that is structurally very similar to an unmodified one, arguing against the hypothesis that remdesivir is a competitive inhibitor of ATP. We characterize the entire mechanism of reaction, finding that viral RdRp is highly processive and displays a higher catalytic rate of incorporation than human RNA Pol II. Overall, our study provides the first detailed explanation of the replication mechanism of RdRp.

A Career in Biocatalysis: Kurt Faber

Kurt Faber, a synthetic organic chemist by training, strongly influenced the field of biocatalysis throughout the course of his four-decade career (from the late 1980s to the early 2020s). The evolution of his career has gone hand in hand with the development of biocatalysis into a mature and versatile discipline originally deep-rooted in organic chemistry that quickly integrated techniques from molecular biology. His work has provided numerous grounds for recognizing natural catalysts as essential tools in organic synthesis, complementing the more chemical approaches. An important aspect of Kurt Faber's career encompasses teaching, mentoring, and encouraging the development of younger scientists worldwide. His book, Biotransformations in Organic Chemistry, now in its seventh edition, has become a recognized tool for training generations of chemists and biotechnologists and remains a reference worldwide. At Biotrans 2021, the 15th International Symposium on Biocatalysis and Biotransformations (originally scheduled to be held in Graz, Austria, and eventually held online due to the COVID-19 pandemic), Kurt Faber received the Biotrans senior award in recognition of his innovative contributions to the field. In this Account, we would like to acknowledge his contributions to the field of biocatalysis, highlighting both the growing importance of this discipline in many (industrial) sectors and the profound changes that have occurred in recent decades, while reflecting on some of Kurt Faber's most important discoveries and his legacy to this field. © 2022 The Authors. Published by American Chemical Society.

Industrially Relevant Enzyme Cascades for Drug Synthesis and Their Ecological Assessment.

Environmentally friendly and sustainable processes for the production of active pharmaceutical ingredients (APIs) gain increasing attention. Biocatalytic synthesis routes with enzyme cascades support many stated green production principles, for example, the reduced need for solvents or the biodegradability of enzymes. Multi-enzyme reactions have even more advantages such as the shift of the equilibrium towards the product side, no intermediate isolation, and the synthesis of complex molecules in one reaction pot. Despite the intriguing benefits, only a few enzyme cascades have been applied in the pharmaceutical industry so far. However, several new enzyme cascades are currently being developed in research that could be of great importance to the pharmaceutical industry. Here, we present multi-enzymatic reactions for API synthesis that are close to an industrial application. Their performances are comparable or exceed their chemical counterparts. A few enzyme cascades that are still in development are also introduced in this review. Economic and ecological considerations are made for some example cascades to assess their environmental friendliness and applicability.

Production of Jet Biofuels by Catalytic Hydroprocessing of Esters and Fatty Acids: A Review

The transition from fossil to bio-based fuels is a requisite for reducing CO2 emissions in the aviation sector. Jet biofuels are alternative aviation fuels with similar chemical composition and performance of fossil jet fuels. In this context, the Hydroprocessing of Esters and Fatty Acids (HEFA) presents the most consolidated pathway for producing jet biofuels. The process for converting esters and/or fatty acids into hydrocarbons may involve hydrodeoxygenation, hydrocracking and hydroisomerization, depending on the chemical composition of the selected feedstock and the desired fuel properties. Furthermore, the HEFA process is usually performed under high H2 pressures and temperatures, with reactions mediated by a heterogeneous catalyst. In this framework, supported noble metals have been preferably employed in the HEFA process;however, some efforts were reported to utilize non-noble metals, achieving a similar performance of noble metals. Besides the metallic site, the acidic site of the catalyst is crucial for product selectivity. Bifunctional catalysts have been employed for the complete process of jet biofuel production with standardized properties, with a special remark for using zeolites as support. The proper design of heterogeneous catalysts may also reduce the consumption of hydrogen. Finally, the potential of enzymes as catalysts for intermediate products of the HEFA pathway is highlighted.

Semi-Automated High-Throughput Substrate Screening Assay for Nucleoside Kinases.

Nucleoside kinases (NKs) are key enzymes involved in the in vivo phosphorylation of nucleoside analogues used as drugs to treat cancer or viral infections. Having different specificities, the characterization of NKs is essential for drug design and nucleotide analogue production in an in vitro enzymatic process. Therefore, a fast and reliable substrate screening method for NKs is of great importance. Here, we report on the validation of a well-known luciferase-based assay for the detection of NK activity in a 96-well plate format. The assay was semi-automated using a liquid handling robot. Good linearity was demonstrated (r² > 0.98) in the range of 0-500 µM ATP, and it was shown that alternative phosphate donors like dATP or CTP were also accepted by the luciferase. The developed high-throughput assay revealed comparable results to HPLC analysis. The assay was exemplarily used for the comparison of the substrate spectra of four NKs using 20 (8 natural, 12 modified) substrates. The screening results correlated well with literature data, and additionally, previously unknown substrates were identified for three of the NKs studied. Our results demonstrate that the developed semi-automated high-throughput assay is suitable to identify best performing NKs for a wide range of substrates.

Applications of Ionic Liquids in Whole-Cell and Isolated Enzyme Biocatalysis.

Ionic liquids have unique chemical properties that have fascinated scientists in many fields. The effects of adding ionic liquids to biocatalysts are many and varied. The uses of ionic liquids in biocatalysis include improved separations and phase behaviour, reduction in toxicity, and stabilization of protein structures. As the ionic liquid state of the art has progressed, concepts of what can be achieved in biocatalysis using ionic liquids have evolved and more beneficial effects have been discovered. In this review ionic liquids for whole-cell and isolated enzyme biocatalysis will be discussed with an emphasis on the latest developments, and a look to the future.

An Expedient Synthesis of Flexible Nucleosides through Enzymatic Glycosylation of Proximal and Distal Fleximer Bases.

The structurally unique "fleximer" nucleosides were originally designed to investigate how flexibility in a nucleobase could potentially affect receptor-ligand recognition and function. Recently they have been shown to have low-to-sub-micromolar levels of activity against a number of viruses, including coronaviruses, filoviruses, and flaviviruses. However, the synthesis of distal fleximers in particular has thus far been quite tedious and low yielding. As a potential solution to this issue, a series of proximal fleximer bases (flex-bases) has been successfully coupled to both ribose and 2'-deoxyribose sugars by using the N-deoxyribosyltransferase II of Lactobacillus leichmannii (LlNDT) and Escherichia coli purine nucleoside phosphorylase (PNP). To explore the range of this facile approach, transglycosylation experiments on a thieno-expanded tricyclic heterocyclic base, as well as several distal and proximal flex-bases were performed to determine whether the corresponding fleximer nucleosides could be obtained in this fashion, thus potentially significantly shortening the route to these biologically significant compounds. The results of those studies are reported herein.