Lipase immobilization via cross-linked enzyme aggregates: Problems and prospects – A review

International Journal of Biological Macromolecules, Journal Year: 2022, Volume and Issue: 215, P. 434 - 449

Published: June 22, 2022

In this review we have focused on the preparation of cross-linked enzyme aggregates (CLEAs) from lipases, as these are among most used in bioprocesses. This immobilization method is considered very attractive due to simplicity, non-use supports and possibility using crude extracts. CLEAs provide lipase stabilization under extreme temperature or pH conditions presence organic solvents, addition preventing leaching aqueous medium. However, it presents some problems limitations their use. The refer mainly crosslinking step, may be solved an aminated feeder. handling been tackled designing magnetic-CLEAs trapping particles with better mechanical properties, substrate diffusion has reduced by producing more porous-CLEAs, etc. co-immobilization combi-CLEAs also a new tendency. Therefore, explores methodology aimed at its applications.

Language: Английский

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Microbial Lipases and Their Potential in the Production of Pharmaceutical Building Blocks

International Journal of Molecular Sciences, Journal Year: 2022, Volume and Issue: 23(17), P. 9933 - 9933

Published: Sept. 1, 2022

Processes involving lipases in obtaining active pharmaceutical ingredients (APIs) are crucial to increase the sustainability of industry. Despite their lower production cost, microbial striking for versatile catalyzing reactions beyond physiological role. In context taking advantage synthesis API building blocks, this review focuses on: (i) structural origins catalytic properties lipases, including results techniques such as single particle monitoring (SPT) and description its selectivity Kazlauskas rule "Mirror-Image Packing" or "Key Region(s) influencing enantioselectivity" (KRIE); (ii) immobilization methods given conferred operative advantages industrial applications modulating capacity lipase properties; (iii) a comprehensive use conventional promiscuous catalyst key organic (Knoevenagel condensation, Morita-Baylis-Hillman (MBH) reactions, Markovnikov additions, Baeyer-Villiger oxidation, racemization, among others). Finally, will also focus on research perspective necessary application development towards greener

Language: Английский

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Cascades of Evolved Enzymes for the Synthesis of Complex Molecules

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(39)

Published: Aug. 26, 2022

Abstract Thanks to advances in enzyme discovery and protein engineering combined with the development of enzymatic multistep reaction cascades, new efficient routes for drug synthesis have been created that are superior chemical syntheses. This supports goal pharmaceutical industries move more sustainable environmentally friendly processes. Recently described outstanding examples include biocatalytic cascade syntheses cyclic dinucleotide MK‐1454, molnupiravir, islatravir, as well fixation CO 2 make starch using an artificial cascade.

Language: Английский

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Bayesian Optimization for an ATP-Regenerating In Vitro Enzyme Cascade

Catalysts, Journal Year: 2023, Volume and Issue: 13(3), P. 468 - 468

Published: Feb. 23, 2023

Enzyme cascades are an emerging synthetic tool for the synthesis of various molecules, combining advantages biocatalysis and one-pot multi-step reactions. However, more complex enzyme cascade is, difficult it is to achieve adequate productivities product concentrations. Therefore, whole process must be optimized account synergistic effects. One way deal with this challenge involves data-driven models in combination experimental validation. Here, Bayesian optimization was applied ATP-producing -regenerating consisting polyphosphate kinases. The co-substrate concentrations were adjusted ATP-dependent reaction, catalyzed by mevalonate kinase (MVK). With a total 16 experiments, we able iteratively optimize initial components used improve specific activity MVK 10.2 U mg−1. even exceeded results reference reaction stoichiometrically added ATP amounts, which 8.8 mg−1 reached. At same time, also improved so that complete yields achieved.

Language: Английский

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Evaluating Multienzyme Cascade Routes for Pharmaceutically Relevant Molecules

European Journal of Organic Chemistry, Journal Year: 2024, Volume and Issue: 27(8)

Published: Jan. 5, 2024

Abstract The constant demand for sustainable approaches to pharmaceutical synthesis has thrived the enzyme catalysis (biocatalysis) field. most sought‐after features like high specificity, environmentally benign, and biodegradable nature have rendered enzymatic processes predominantly eco‐friendly. Owing these merits, biocatalysis is now being developed extensively exploit their applications in designing alternate synthetic routes manufacturing pharmaceuticals, line with green chemistry principles. Further, combining multiple enzymes one‐pot a highly efficient methodology as it eliminates intermediate isolation, shifts equilibrium forward direction, proceeds ambient aqueous conditions. This review highlights importance of recently multienzyme cascade pharmaceutically important bioactive molecules by relating them well‐known chemical route.

Language: Английский

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Window of sustainable bioprocess operation: towards merging environmental sustainability assessment and process operation at early-stage bioprocess development

Current Opinion in Biotechnology, Journal Year: 2025, Volume and Issue: 93, P. 103292 - 103292

Published: March 18, 2025

Language: Английский

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Biological activity of new bioactive steroids deriving from biotransformation of cortisone

Microbial Cell Factories, Journal Year: 2022, Volume and Issue: 21(1)

Published: Nov. 24, 2022

Abstract Cortisone is a metabolite belonging to the corticosteroid class that used pharmaceutically directly as drug or prodrug. In addition its large consumption, use linked several side effects, so pharmaceutical research aims develop effective drugs with low no alternative compounds cortisone are part of an active investment in ongoing on discovery. Since biotransformation can be considered source new molecules potential therapeutic use, present work focuses preliminary vitro study aimed at evaluating mutagenic, anti-inflammatory, antioxidant and neuroprotective activity SCA SCB obtained from using Rh. Rhodnii strain DSM 43960. The results very encouraging due safety biotransformed reference genotoxicity checked by Ames test, high capacity anti-inflammatory activity. fact, thecompounds inhibited both TNFα-stimulated expression secretion NFkB target cytokines, COX activity, activate glucocorticoid receptor. Finally exhibited properties.

Language: Английский

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Debottlenecking of an In Vitro Enzyme Cascade Using a Combined Model‐ and Experiment‐Based Approach

Chemie Ingenieur Technik, Journal Year: 2023, Volume and Issue: 95(4), P. 543 - 548

Published: Feb. 10, 2023

Abstract Designing a feasible in vitro enzyme cascade is challenging task that often preceded by optimization steps. Computational methods can increase the efficiency of process providing deeper insight into system. In this study, bottlenecks farnesyl pyrophosphate‐producing were determined combination silico modeling and wet‐lab experiments. ATP precursor‐isomerizing isopentenyl diphosphate δ ‐isomerase identified as limitations cascade.

Language: Английский

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Pushing the boundaries of phosphorylase cascade reaction for cellobiose production I: Kinetic model development

Biotechnology and Bioengineering, Journal Year: 2023, Volume and Issue: 121(2), P. 580 - 592

Published: Nov. 20, 2023

Abstract One‐pot cascade reactions of coupled disaccharide phosphorylases enable an efficient transglycosylation via intermediary α‐ d ‐glucose 1‐phosphate (G1P). Such transformations have promising applications in the production carbohydrate commodities, including cellobiose for food and feed use. Several studies shown sucrose phosphorylase synthesis from sucrose, but boundaries on transformation efficiency that result kinetic thermodynamic characteristics individual enzyme are not known. Here, we assessed a step‐by‐step systematic fashion practical requirements model to describe at industrially relevant substrate concentrations up 600 mM glucose each. Mechanistic initial‐rate models two‐substrate (sucrose + phosphate → G1P fructose) (G1P phosphate) were needed additionally required expansion by terms inhibition, particular distinctive two‐site inhibition (from Cellulumonas uda ). Combined with mass action accounting approach equilibrium, gave excellent fit robust prediction full reaction time courses wide range activities as well concentrations, variable substoichiometric concentration phosphate. The thus provides essential engineering tool disentangle highly interrelated factors conversion reaction; it establishes necessary basis window operation calculations targeted optimizations toward different process tasks.

Language: Английский

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Pushing the boundaries of phosphorylase cascade reaction for cellobiose production II: Model‐based multiobjective optimization

Biotechnology and Bioengineering, Journal Year: 2023, Volume and Issue: 121(2), P. 566 - 579

Published: Nov. 21, 2023

Abstract The inherent complexity of coupled biocatalytic reactions presents a major challenge for process development with one‐pot multienzyme cascade transformations. Kinetic models are powerful engineering tools to guide the optimization towards performance suitable scale up an actual production. Here, we report kinetic model‐based window operation analysis cellobiose production (≥100 g/L) from sucrose and glucose by indirect transglycosylation via 1‐phosphate as intermediate. two‐step transformation is catalyzed phosphorylase in presence substoichiometric amounts phosphate (≤27 mol% substrate). modeling was instrumental uncover hidden effect bulk microviscosity due high sugar concentrations on decreasing rate specifically. mechanistic‐empirical hybrid model thus developed gives comprehensive description reaction at industrially relevant substrate conditions. Model simulations serve unravel opposed relationships between efficient utilization enzymes maximized concentration (or yield) product within given time, dependence initial used. Optimum balance these competing key metrics suggested model‐calculated verified experimentally. evidence shown highlights important use characterization ways that appear be inaccessible purely data‐driven approaches.

Language: Английский

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