Biocatalysis

Using biocatalysts for chemical manufacturing is a highly developed and applied science, used widely for production and processing of pharmaceuticals and food items. Both enzymes and whole cells can be used to catalyse organic reactions, with the most general example of biocatalysis being the yeast-mediated transformation of sugars into alcohols, acids or gases.

However, the most frequently employed group of biocatalysts is lipases. These enzymes are very versatile, facilitating the hydrolysis of lipids into fatty acids as well as the synthesis of esters and amides. Areas of lipase-catalysed reactions include food processing, synthesis of fine chemicals and production of biofuel. Other regularly used enzymatic biocatalysts are proteases, cellulases and amylases, used for the hydrolysis of proteins, cellulose and starch, respectively

The technique of using enzymes to mediate organic synthesis is widely endorsed due to their high selectivity and specificity, which ensures a more satisfactory yield and purity of the transformed products. Furthermore, by the use of protein engineering, the biocatalysis can be made even more efficient by having the enzyme customized to fit a specific need

Immobilized enzymes

When utilizing isolated enzymes for organic synthesis, a typical approach is to keep the enzymes immobilized on the surface of an insoluble material. This is preferred to using soluble enzymes due to both technical and economic benefits. The immobilization will allow for easier recycling, manipulation and separation of the enzyme, as well as an increase in its thermal and operational stability. Additionally, immobilization of the enzyme will ensure minimized protein contamination during the experimental procedure, and improved control over the overall reaction process.

Rotating bed reactors

A rotating bed reactor (RBR) is designed to prevent disintegration of solid phase particles by retaining them inside a rotating cylinder during the reaction span. This minimizes solid phase debris even at high speeds, which would normally occur due to mechanical forces when using stirred tank reactors. Due to these properties of the RBR, reaction rates can be improved by increasing the speed at which an RBR is rotated, without shortening the lifespan of the solid phase particles. This ensures easy in-line monitoring of the process, as well as a more efficient recycling of the solid phase particles, making the RBR a cost and time efficient alternative to conventional methods for immobilized enzyme biocatalysis.

Application 1014
Biocatalysis by immobilized enzymes in a rotating bed reactor

Time lapse video showing how straightforward it is to use immobilized enzymes in a rotating bed reactor. A substrate giving a yellow coloured product was used to follow the reaction progress of an ester hydrolysis by an immobilized lipase. This substrate is commonly used to screen and characterize lipases.

Application L1704
Biocatalysis engineering: the big picture

A tutoral review taking a holistic approach to the engineering of biotransformations utilising isolated enzymes. The authors highlighted SpinChem® RBR technology benefits and wrote; “… in addition to protecting the biocatalyst from shear forces caused by the stirrer, it greatly accelerates mass transfer, thus affording substantially higher reaction rates and creating the possibility to use much smaller reactors”.   Keywords: Biotransformation, Immobilized enzymes, Organic molecules, Review, Scientific literature

Application 1023
Biocatalysis in rotating bed reactors – from screening to production

Poster describing the screening of resins with immobilized enzymes for esterification reactions. The screening was performed with prepacked cartridges inside a SpinChem® rotating bed reactor (RBR) and compared to parallel screening with prototype disposable magnetic RBR (MagRBR) in 10 mL volume. Both approaches were very quick, required minimum preparation and no filtration. Up-scaled processes enabled quick enzyme recycling, opening for the possibility of semi-continuous processes with attractive production economy.

Application 1003
Effective phase-transfer between immiscible liquids and an ion exchange resin

Illustrative video showing how a phenolic colourant is deprotonated and extracted from an organic to an aqueous solvent. Using SpinChem® RBR in a flower-baffled reaction vessel created fine emulsion droplets resulting in effective phase-transfer between the two liquids and the solid phase. Keywords: Immiscible liquids, Ion exchange, Organic molecules, Technology

Application L1301
Efficient biocatalysis with immobilized enzymes or encapsulated whole cell microorganism by using the SpinChem reactor system

Hendrik Mallin, Jan Muschiol, Dr. Emil Byström, Prof. Dr. Uwe T. BornscheuerChemCatChem, 5 (2013) 3529-3532   "...the immobilized transaminase was better protected from mechanical forces in the SpinChem device."    Keywords: Alginate, Biotransformation, Encapsulated cells, Immobilized enzymes, Organic molecules, Scientific literature

Application 1002
Efficient synthesis of chiral lactones by encapsulated cells in a rotating bed reactor

Comparison of SpinChem® rotating bed reactor (RBR) with traditional reaction set-ups for a demanding biotransformation. SpinChem® RBR matched or outperformed the other systems and gave a 10 to 25-fold more time-efficient recycling of the encapsulated cells.   Keywords: Alginate, Biotransformation, Encapsulated cells, Gas-distribution, Organic molecules, Quick recycling

Application 1028
Enzyme immobilization screening using magnetic rotating bed reactors

The SpinChem® MagRBR ECR screening kit, pre-packed with Purolite® Lifetech™ resins, was used to screen six different enzyme carrier resins in parallel for the immobilization of lipase CalB. Easy sampling and monitoring of the process, together with effortless handling, established the MagRBR as a time and labour efficient screening device. Keywords: Biotransformation, Easy handling, Immobilized enzymes, Rapid screening

Application L1402
Lipase catalyzed regioselective lactamization as a key step in the synthesis of N-Boc (2R)-1,4-oxazepane-2-carboxylic acid

A synthesis of N-Boc (2R)-1,4-oxazepane-2-carboxylic acid was developed in 39% yield over seven steps starting from methyl (2R)-glycidate. The key step was a lipase-catalyzed regioselective lactamization performed using SpinChem® rotating bed reactor that simplified work up and recycling of the enzyme. Carl-Johan Aurell*, Staffan Karlsson, Fritiof Pontén, and Søren M. Andersen Keywords: Biotransformation, Immobilized enzymes, Organic molecules, Quick recycling, Scientific literature

Application L1602
Modularized biocatalysis: Immobilization of whole cells for preparative applications in microaqueous organic solvents

Jochen Wachtmeister, Philip Mennicken, Andreas Hunold, Dörte RotherChemCatChem, 8 (2016) 607-614   Keywords: Biotransformation, Encapsulated cells, Organic molecules, Scientific literature

Application L1703
Reaction engineering of biocatalytic (S)-naproxen synthesis integrating in-line process monitoring by Raman spectroscopy

A traditional stirred tank reactor setup was compared to a rotating bed reactor (RBR) for the biocatalytic synthesis of the anti-inflammatory drug (S)-naproxen. Both setups performed well during five repetitive bathes giving an enantiomeric excess of 99% and an isolated yield of 92%, but the RBR was easier to handle and the authors concluded that “… the rotating bed reactor concept can be regarded as a promising option for industrial applications”. Keywords: Biotransformation, Immobilized enzymes, Organic molecules, Scientific literature

Application L1606
Recent advances in whole cell biocatalysis techniques bridging from investigative to industrial scale

Jochen Wachtmeister, Dörte RotherCurrent Opinion in Biotechnology, 42 (2016) 169-177 Keywords: Biotransformation, Encapsulated cells, Organic molecules, Scientific literature

Application 9001
Recycling of Immobilized Enzymes – Simple and Streamlined Biocatalysis

Immobilized catalyst recycling using a SpinChem® rotating bed reactor (RBR) and a Mettler-Toledo EasyMax™ 102 Advanced synthesis workstation. The process proved very time efficient as no filtration steps were needed between cycles, or for the samples extracted for analysis during each run. Washing of the resin between runs was fast, simple and robust, without running the risk of material loss. Keywords: Biotransformation, Immobilized enzymes, Mettler-Toledo, Organic molecules, Preserved activity, Quick recycling

Application 1032
Rotating bed reactor for immobilized enzymatic reactions

Poster on a case study of applying the rotating bed reactor for the lipase-mediated stereoselective acetylation of a racemate amine as a model reaction for the manufacturing of pharmaceutical building blocks. The results showed that enzyme recycling and synthesis scale up was easy to achieve with preserved yield, enantioselectivity and catalytic activity.   Keywords: Biotransformation, Easy handling, Immobilized enzymes, Quick recycling, Seamless scaleup

Application 1025
Rotating bed reactor setup for in situ formation, loading and handling of alginate beads and whole cell encapsulation

Video showing the formation of alginate beads under conditions mimicking whole cell encapsulation. The use of a SpinChem® rotating bed reactor (RBR) allowed easy collection, maturing and washing of the alginate beads. With the RBR setup, it was possible to immediately continue with filling of the reaction substrate into the same vessel, thus reducing the number of handling steps and facilitating bead recycling. The beads showed no signs of physical wear after use in the RBR.   Keywords: Alginate, Biotransformation, Easy handling, One-pot multistep, Technology

Application 9002
Screening of Immobilized Enzymes – Fast and Convenient Reaction Optimization

The stable reaction environment in the EasyMax™ 102 Advanced synthesis workstation and the high flow rates through the SpinChem® RBR allowed for quick and convenient screening of different immobilized lipases to find the enzyme most suitable for further reaction optimization.   Keywords: Biotransformation, Immobilized enzymes, Mettler-Toledo, Organic molecules, Rapid screening

Application 1027
Screening of immobilized lipases using magnetic rotating bed reactors

Six different immobilized lipases were screened in parallel for the esterification of lauric acid into propyl laurate using the pre-packed SpinChem® MagRBR Lipase screening kit. The process proved fast and simple, as efficient sampling and monitoring of the process was achieved without filtration steps, by keeping the immobilized catalyst confined inside the MagRBR.   Keywords: Biotransformation, Easy handling, Immobilized enzymes, Organic molecules, Rapid screening

Application 1009
Simultaneous extraction of two dyes selectively onto different resins

Video illustrating how a mixture of red and blue dyes with different chemical properties can be selectively extracted onto different adsorbents within the same run using a rotating bed reactor (RBR). The dyes were separated based on ionic and hydrophobic interactions, respectively.  Keywords: Cleantech, Decolouration, Extraction, Ion exchange, One-pot multistep, Organic molecules, Polymeric resin, Technology

Application L2001
Using Spinchem Rotating Bed Reactor Technology for Immobilized Enzymatic Reactions: A Case Study

Subhash Pithani, Staffan Karlsson, Hans Emtenäs, and Christopher T. ÖbergOrganic Process Research & Development 2019 23 (9), 1926-1931 "By using SpiChem RBR technology, we found that recycling of the immobilized enzyme was easy with preserved enantioselectivity and catalytic activity. The final optimized process was successfully demonstrated on a 1 kg scale with 39% isolated yield and 98.8% enantiomeric purity."     Keywords: Scientific literature

Application L1706
Whole-cell cascade biotransformations for one-pot multistep organic synthesis

This scientific minireview highlights that “one-pot multi-catalysis reactions is a revolutionary tool for multistep synthesis” and that “reaction engineering is indispensable to further advance biotransformation processes”. The authors concluded that “advanced reactor design such as … the SpinChem reactor, could also enhance cascade biotransformation processes”.   Keywords: Biotransformation, Encapsulated cells, One-pot multistep, Review, Scientific literature

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