Sustainable synthesis of L-phenylalanine derivatives in continuous flow by immobilized phenylalanine ammonia lyase

The application of phenyl ammonia lyases for the amination of a variety of cinnamic acids has been shown to be a cost-efficient method to produce a variety of phenylalanine analogues. Nonetheless, as many other biocatalytic tools, the process intensification, especially due to the high equivalents of ammonia needed, and the cost-efficiency of the catalyst production and use have been key points to further prove their usefulness. Here, we investigated the use of previously characterized PALs (AvPAL and PbPAL) for the amination of a series of substituted cinnamic acids. To enhance the process scalability and the reusability of the catalyst, we investigated the use of covalent immobilization onto commercially available supports, creating a heterogeneous catalyst with good recovered activity (50%) and excellent stability. The immobilized enzyme was also incorporated in continuous flow for the synthesis of 3-methoxy-phenyl alanine and 4-nitro-phenylalanine, which allowed for shorter reaction times (20 mins of contact time) and excellent conversions (88 ± 4% and 89 ± 5%) respectively, which could be maintained over extended periods of time, up to 24h. This work exemplifies the advantages that the combination of enzyme catalysis with flow technologies can have not only in the reaction kinetics, but also in the productivity, catalyst reusability and downstream processing

Evolution of Biocatalysis at Novartis over the last 40 Years

The fortieth anniversary of biocatalysis started at Ciba-Geigy and later at Novartis is a great time to pause and reflect on development of science and technology in this field. Enzyme-based synthesis became a highly valued enabling tool for pharmaceutical research and development over the last decades. In this perspective we aim to discuss how the scientific approaches and trends evolved over the time and present future challenges and opportunities

Nitrile biotransformationby aspergillus niger

A nitrile-converting enzyme activity was induced in Aspergillus niger K10 by 3-cyanopyridine. The whole cell biocatalyst was active at pH 3–11 and hydrolyzed the cyano group into acid and/or amide functions in benzonitrile as well as in its meta- and para-substituted derivatives, cyanopyridines, 2-phenylacetonitrile and thiophen-2-acetonitrile. Amides constituted a significant part of the total biotransformation products of 2- and 4-cyanopyridine, 4-chlorobenzonitrile, 4-tolunitrile and 1,4-dicyanobenzene, while α-substituted acrylonitriles gave amides as the sole product

Bioinformatic Toolbox for the Development of novel CH-Activation Biocatalysts (CH-Pred)

Enzymes are particularly well-suited catalysts for the site-specific C-H activation of active pharmaceutical intermediates. To accelerate enzyme development, an in silico screening workflow for the identification of active CH-activation enzymes will be developed and applied in product synthesis

BM CDT Research Proposals 2018/19 from GDC BIR. Title: Stereoselective, scalable route towards phosphorothioate analogues of ATP

Enantioselective synthesis of phosphorothioate analogues of adenosine 5'-triphosphate is difficult to achieve in highly atom and cost-efficient way.1 Novel ideas to address this challenge are sought after, which could lead to a scalable and sustainable route towards analogues of (Sp)-ATP-αS

Enzymatic Bioconjugation: A Perspective from the Pharmaceutical Industry.

Enzymes have firmly established themselves as bespoke catalysts for small molecule transformations in the pharmaceutical industry, from early research and development stages to large-scale production. In principle, their exquisite selectivity and rate acceleration can also be leveraged for modifying macromolecules to form bioconjugates. However, available catalysts face stiff competition from other bioorthogonal chemistries. In this Perspective, we seek to illuminate applications of enzymatic bioconjugation in the face of an expanding palette of new drug modalities. With these applications, we wish to highlight some examples of current successes and pitfalls of using enzymes for bioconjugation along the pipeline and try to illustrate opportunities for further development

The Development of Biocatalysis as a Tool for Drug Discovery

Enzymes are versatile biocatalysts capable of performing selective reactions. The advantages of enzymes in comparison to classical chemistry including chemical catalysts are the generally milder process conditions and avoidance of harmful reactants. Their high selectivity and specificity are especially beneficial for the enzymatic synthesis of new products with potential applications in drug research. Therefore, in the past decades, the utilization of isolated enzymes or whole-cell biocatalysts has spread through a growing number of biotechnological industries. The applications comprise the production of chiral building blocks for the pharmaceutical and fine chemical industry, the enzymatic synthesis of drug metabolites for testing of toxicity, function, biological activity, degradation and the production of biocatalytically modified natural products, which all play a role in drug discovery. Especially Oreste Ghisalba's contributions, which paved the way for the industrial use of enzymes, will be considered in this review

Discovery of novel Fe(II)/α-ketoglutarate-dependent dioxygenases for oxidation of L-proline

Genome-mining for novel Fe(II)/α-ketoglutarate-dependent dioxygenases (αKGDs) to expand the enzymatic repertoire in the oxidation of L-proline is reported. Through clustering of genes, we predicted regio- and stereoselectivity in hydroxylation reaction and validated this hypothesis experimentally. Two novel by-products in reactions with BcePH and Ssp5PH were observed, isolated and structure was determined as an epoxide and a 3,4-diol, respectively. Mechanism for formation of epoxide is suggested and validated by using 18O-labelling experiment, that proceeds via cis-3-hydroxylation step first, followed by ring closure. A Biocatalytic step was performed on sub-gram quantities of starting material without any significant condition optimization. The substrate concentration, however, is already up to 40-fold higher than the usually reported titers for P450-mediated hydroxylations, showing the synthetic potential of αKGDs on preparative scal

Surfing the biocatalysis wave to new applications Changed to: From nature to industry: Harnessing enzymes for biocatalysis, as per request of Editor

Biocatalysis applies enzymes to make valuable products. Today, this green technology is effectively utilized from bench scale to industrial production. The last decade has seen an explosion in the development of experimental and in silico tools to tailor enzymatic properties, the latter relying on the surge of available bioinformatic data and continuously accelerating computational advances. By harnessing (chemo)-enzymatic synthesis routes or intricate enzyme cascades, completely new targets can be synthesized, ranging from DNA and complex pharmaceuticals to starch made in vitro simply from CO2-derived methanol. In addition, intriguing new chemistries have emerged by combining biocatalysis with transition metal-, photo- and electrocatalysis. Against this exciting backdrop, this review highlights recent key developments, identifies current limitations, and provides a future prospect for this rapidly developing research field