Immobilizing Systems Biocatalysis for the Selective Oxidation of Glycerol Coupled to In Situ Cofactor Recycling and Hydrogen Peroxide Elimination

Rocha-Martín, Javier; Acosta, Andreína; Guisán, José M.; López‐Gallego, Fernando

doi:10.1002/cctc.201500210

Cited by 25 publications

(25 citation statements)

References 40 publications

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“…A tri-enzyme system for selective oxidation of glycerol was improved, but in this case it was necessary to combine protein mutagenesis and immobilization techniques. The three enzymes were co-immobilized sequentially in the same porous carrier and co-crosslinked [21]. A bi-enzymatic system consisting of L-N-carbamoylase from Geobacillus stearothermophilus CECT43 (BsLcar) and N-succinyl-amino acid racemase from Geobacillus kaustophilus CECT 4264 (GkNSAAR) as carbamoyl racemase was immobilized in Immobeads IB-161, showing 75% activity after 10 reaction cycles.…”

Section: Efficiency Of the System: Substrate Spectra And Reaction Cyclesmentioning

confidence: 99%

l-Amino Acid Production by a Immobilized Double-Racemase Hydantoinase Process: Improvement and Comparison with a Free Protein System

et al. 2017

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Protein immobilization is proving to be an environmentally friendly strategy for manufacturing biochemicals at high yields and low production costs. This work describes the optimization of the so-called "double-racemase hydantoinase process," a system of four enzymes used to produce optically pure L-amino acids from a racemic mixture of hydantoins. The four proteins were immobilized separately, and, based on their specific activity, the optimal whole relation was determined. The first enzyme, D,L-hydantoinase, preferably hydrolyzes D-hydantoins from D,L-hydantoins to N-carbamoyl-D-amino acids. The remaining L-hydantoins are racemized by the second enzyme, hydantoin racemase, and continue supplying substrate D-hydantoins to the first enzyme. N-carbamoyl-D-amino acid is racemized in turn to N-carbamoyl-L-amino acid by the third enzyme, carbamoyl racemase. Finally, the N-carbamoyl-L-amino acid is transformed to L-amino acid by the fourth enzyme, L-carbamoylase. Therefore, the product of one enzyme is the substrate of another. Perfect coordination of the four activities is necessary to avoid the accumulation of reaction intermediates and to achieve an adequate rate for commercial purposes. The system has shown a broad pH optimum of 7-9, with a maximum activity at 8 and an optimal temperature of 60 • C. Comparison of the immobilized system with the free protein system showed that the reaction velocity increased for the production of norvaline, norleucine, ABA, and homophenylalanine, while it decreased for L-valine and remained unchanged for L-methionine.

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Section: Efficiency Of the System: Substrate Spectra And Reaction Cyclesmentioning

confidence: 99%

l-Amino Acid Production by a Immobilized Double-Racemase Hydantoinase Process: Improvement and Comparison with a Free Protein System

et al. 2017

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“…The colocalisation of both enzymes across the same solid surface normally favours improved performance of the reaction cascade due to channelling of substrates and cofactors between the different enzyme active sites. In this case, control of the spatial organisation of each enzyme is compulsory to enhance the kinetics and performance of the system …”

Section: Introductionmentioning

confidence: 99%

Sustainable and Continuous Synthesis of Enantiopure l‐Amino Acids by Using a Versatile Immobilised Multienzyme System

et al. 2017

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The enzymatic synthesis of α-amino acids is a sustainable and efficient alternative to chemical processes, through which achieving enantiopure products is difficult. To more address this synthesis efficiently, a hierarchical architecture that irreversibly co-immobilises an amino acid dehydrogenase with polyethyleneimine on porous agarose beads has been designed and fabricated. The cationic polymer acts as an irreversible anchoring layer for the formate dehydrogenase. In this architecture, the two enzymes and polymer colocalise across the whole microstructure of the porous carrier. This multifunctional heterogeneous biocatalyst was kinetically characterised and applied to the enantioselective synthesis of a variety of canonical and noncanonical α-amino acids in both discontinuous (batch) and continuous modes. The co-immobilised bienzymatic system conserves more than 50 % of its initial effectiveness after five batch cycles and 8 days of continuous operation. Additionally, the environmental impact of this process has been semiquantitatively calculated and compared with the state of the art.

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“…Few research works refer to a multi‐step reaction with immobilized proteins and this study may therefore be a promising incentive for industrial applications in biocatalysis . An enzymatic cascade implies continuous substrate feedback and free mass transfer.…”

Section: Introductionmentioning

confidence: 99%

“…An enzymatic cascade implies continuous substrate feedback and free mass transfer. The different enzyme activities must be perfectly coordinated as they are working in an environment that is so different to the natural media . In this situation the use of the four enzymes in solution proved successful but highly expensive.…”

Section: Introductionmentioning

confidence: 99%

“…Few research works refer to a multi-step reaction with immobilized proteins and this study may therefore be a promising incentive for industrial applications in biocatalysis. 7 -10 An enzymatic cascade implies continuous substrate feedback and free mass transfer. The different enzyme activities must be perfectly coordinated as they are working in an environment that is so different to the natural media.…”

Section: Introductionmentioning

confidence: 99%

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Immobilization of a multi‐enzyme system for L‐amino acids production

Rodríguez-Alonso

Rodríguez‐Vico

Heras‐Vázquez

et al. 2015

J of Chemical Tech & Biotech

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BACKGROUND Four enzymes were immobilized for the production of optically pure natural and non‐natural L‐amino acids via the ‘Double‐Racemase Hydantoinase Process’. Immobilization constitutes an empirical process, and for each enzyme we tested 11 carriers with different functional groups; epoxide, EC‐EP, EC‐HFA, IB‐150 and IB‐350, carboxylic acid IB‐C435, quaternary ammonium IB‐A161, IB‐A171 and IB‐A369, aromatic group IB‐S861, and hydroxyl group IB‐S60S and IB‐S60P. RESULTS Each protein showed preference for binding on one or several supports: D,L‐hydantoinase/IB‐350, hydantoin racemase/EC‐EP, L‐carbamoylase/EC‐EP and carbamoyl racemase/IB‐A161. The process was optimized for each enzyme by modifying temperature, pH and ionic strength. For the enzymatic cascade, it was demonstrated that it was essential to use supports having homogeneous characteristics. The product of the first reaction is the substrate of the next one, and so on. Free mass diffusion from one enzyme to the other is crucial to avoid retention on the support and to maintain the protein–substrate interaction constant. CONCLUSION It was proved that support size, weight and hydrophobicity must be homogeneous to avoid the formation of separate layers of the matrix selected. For this reason, the support IB‐350 was used for the four enzymes, even though it may not be the most efficient one for some of them. © 2015 Society of Chemical Industry

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Immobilizing Systems Biocatalysis for the Selective Oxidation of Glycerol Coupled to In Situ Cofactor Recycling and Hydrogen Peroxide Elimination

Cited by 25 publications

References 40 publications

l-Amino Acid Production by a Immobilized Double-Racemase Hydantoinase Process: Improvement and Comparison with a Free Protein System

l-Amino Acid Production by a Immobilized Double-Racemase Hydantoinase Process: Improvement and Comparison with a Free Protein System

Sustainable and Continuous Synthesis of Enantiopure l‐Amino Acids by Using a Versatile Immobilised Multienzyme System

Immobilization of a multi‐enzyme system for L‐amino acids production

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