Biocatalytic production of semi-synthetic cephalosporins: process technology and integration

Tramper, J.; Beeftink, H.H.; Janssen, A.E.M.; Ooijkaas, L.P.; Roon, J.L. van; Strubel, M.; Schroën, C.G.P.H.

doi:10.1007/978-94-010-0850-1_5

Cited by 5 publications

(4 citation statements)

References 33 publications

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“…These include facilitated biocatalyst reuse due to high mechanical and operational (apparent) stability, the possibility to operate at extremely high biocatalyst loadings, leading to small reactor sizes, and the possibility to employ the biocatalyst in packed‐bed or fluidized‐bed reactors. In this way, immobilization may lead to efficient (possibly continuous) conversions with high volumetric productivity and a substantial half‐life time (1, 2).…”

Section: Introductionmentioning

confidence: 99%

Enzyme Distribution Derived from Macroscopic Particle Behavior of an Industrial Immobilized Penicillin‐G Acylase

Roon¹,

Joerink²,

Rijkers³

et al. 2003

Biotechnology Progress

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The macroscopic kinetic behavior of an industrially employed immobilized penicillin-G acylase, called Assemblase, formed the basis for a discussion on some simple intraparticle biocatalytic model distributions. Assemblase catalyzes the synthesis of the widely used semisynthetic antibiotic cephalexin. Despite the obvious advantages of immobilization, less cephalexin and more of the unwanted byproduct d-(-)-phenylglycine are obtained due to diffusional limitations when the immobilized enzyme is employed. To rationally optimize Assemblase, the parameters particle size, enzyme loading, and enzyme distribution, which severely determine the macroscopic particle performance, were studied on the basis of macroscopic observations. Laser diffraction measurements showed that the particle sizes in Assemblase vary as much as 100-fold. The relative and total enzyme loadings in Assemblase and fractions thereof of different sizes were determined by initial-rate d-(-)-phenylglycine amide hydrolysis, cephalexin synthesis experiments, and active-site titration. These experiments revealed that the loading of penicillin-G acylase in Assemblase was inversely correlated with the particle diameter. Apart from enzyme loadings, estimates on the intraparticle enzyme distribution came from cephalexin synthesis experiments, where mass-transport limitations were present. Although this method cannot provide the level of detail of specific labeling experiments, it is simple, fast, and cheap. Within the set of simple model predictions, a heterogeneous enzyme distribution with most biocatalyst present in the outer region of the particle (within the outer 100 microm) gave the best description of the observed behavior, although no exact correlation was established. Highly detailed determination of intraparticle enzyme distributions must come from immunolabeling.

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Section: Introductionmentioning

confidence: 99%

Enzyme Distribution Derived from Macroscopic Particle Behavior of an Industrial Immobilized Penicillin‐G Acylase

Roon¹,

Joerink²,

Rijkers³

et al. 2003

Biotechnology Progress

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“…419 Hence, the biocatalytic route became an attractive alternative. 18 It reduced the reaction steps (from three to one) as well as the process costs, and it only required ammonia to control the pH (0.09 kg per kg of 6-APA) 420 to neutralize the formed acid. 51 Covalently immobilized acylase is nowadays used to produce >10,000 tons per year of penicillin and cephalosporin antibiotics.…”

Section: Aminesmentioning

confidence: 99%

“…Until about 1970, 6-APA was produced chemically involving protection and deprotection steps, at low temperatures (−40 °C) and at the expense of environmentally problematic reagents (PCl 5 ) and solvents (CH 2 Cl 2 ), which generated huge amounts of waste (10–100 kg of waste per kg of product) . Hence, the biocatalytic route became an attractive alternative . It reduced the reaction steps (from three to one) as well as the process costs, and it only required ammonia to control the pH (0.09 kg per kg of 6-APA) to neutralize the formed acid .…”

Section: Functional Groups Formed In Biocatalytic Reactions For Api S...mentioning

confidence: 99%

“…The use of enzymes for the catalytic conversion of non-natural compounds, denoted as “biotransformation”, was long regarded as a lab-curiosity before it now became a highly valuable tool in synthesis with even further yet unexploited potential to revolutionize the way APIs are made. Most of the early studies employed uncharacterized crude protein extracts or whole microbial cells like in a black-box approach. − A few landmark-innovations are rare prominent exceptions: examples are for instance (i) the synthesis of (−)-ephedrine from benzaldehyde by baker’s yeast fermentation in 1921, , (ii) the Reichstein-process for the production of vitamin C in 1933, employing whole bacterial cells for the regioselective mono-oxidation of sorbose, (iii) the use of penicillin acylase for the production of penicillin derivatives such as cephalosporins during the late 1960s, , and (iv) the regio- and stereoselective hydroxylation of progesterone , enabling the economic production of cortisone.…”

Section: Introductionmentioning

confidence: 99%

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Shortening Synthetic Routes to Small Molecule Active Pharmaceutical Ingredients Employing Biocatalytic Methods

et al. 2021

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Biocatalysis, using enzymes for organic synthesis, has emerged as powerful tool for the synthesis of active pharmaceutical ingredients (APIs). The first industrial biocatalytic processes launched in the first half of the last century exploited whole-cell microorganisms where the specific enzyme at work was not known. In the meantime, novel molecular biology methods, such as efficient gene sequencing and synthesis, triggered breakthroughs in directed evolution for the rapid development of process-stable enzymes with broad substrate scope and good selectivities tailored for specific substrates. To date, enzymes are employed to enable shorter, more efficient, and more sustainable alternative routes toward (established) small molecule APIs, and are additionally used to perform standard reactions in API synthesis more efficiently. Herein, large-scale synthetic routes containing biocatalytic key steps toward >130 APIs of approved drugs and drug candidates are compared with the corresponding chemical protocols (if available) regarding the steps, reaction conditions, and scale. The review is structured according to the functional group formed in the reaction.

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A multicomponent reaction–diffusion model of a heterogeneously distributed immobilized enzyme

Roon

Arntz

Kallenberg

et al. 2006

Appl Microbiol Biotechnol

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A physical model was derived for the synthesis of the antibiotic cephalexin with an industrial immobilized penicillin G acylase, called Assemblase. In reactions catalyzed by Assemblase, less product and more by-product are formed in comparison with a free-enzyme catalyzed reaction. The model incorporates reaction with a heterogeneous enzyme distribution, electrostatically coupled transport, and pH-dependent dissociation behavior of reactants and is used to obtain insight in the complex interplay between these individual processes leading to the suboptimal conversion. The model was successfully validated with synthesis experiments for conditions ranging from heavily diffusion limited to hardly diffusion limited, including substrate concentrations from 50 to 600 mM, temperatures between 273 and 303 K, and pH values between 6 and 9. During the conversion of the substrates into cephalexin, severe pH gradients inside the biocatalytic particle, which were previously measured by others, were predicted. Physical insight in such intraparticle process dynamics may give important clues for future biocatalyst design. The modular construction of the model may also facilitate its use for other bioconversions with other biocatalysts.

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Biocatalytic production of semi-synthetic cephalosporins: process technology and integration

Cited by 5 publications

References 33 publications

Enzyme Distribution Derived from Macroscopic Particle Behavior of an Industrial Immobilized Penicillin‐G Acylase

Enzyme Distribution Derived from Macroscopic Particle Behavior of an Industrial Immobilized Penicillin‐G Acylase

Shortening Synthetic Routes to Small Molecule Active Pharmaceutical Ingredients Employing Biocatalytic Methods

A multicomponent reaction–diffusion model of a heterogeneously distributed immobilized enzyme

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