Distribution of an enzyme in porous polymer beads

Ampon, Kamaruzaman Hj.

doi:10.1002/jctb.280550213

Cited by 11 publications

(3 citation statements)

References 24 publications

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“…In contrast, RhB–LovD–BuCh2 colonized deeper regions of EziG1 resulting in a higher protein infiltration distance (24.42 μm) (Figure d). Additional electrostatic interactions between the enzyme and the EziG1 carrier might contribute to localize the enzyme in inner regions of the porous carrier surface as observed for other enzymes. , Additionally, the larger infiltration within the material is translated into a lower enzyme density per μm 3 of the carrier, thus reducing the protein molecular packing, which could explain the superior rRA of LovD–BuCh2@EziG1 (Table ).…”

Section: Resultsmentioning

confidence: 98%

Immobilization and Stabilization of an Engineered Acyltransferase for the Continuous Biosynthesis of Simvastatin in Packed-Bed Reactors

García‐Marquina

Langer

Sánchez-Costa

et al. 2022

ACS Sustainable Chem. Eng.

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Simvastatin is a top-selling cholesterol-lowering drug traditionally obtained through a semi-synthetic process starting from lovastatin. However, this process is cost-demanding and makes use of chemical reagents that can generate considerable waste. The sustainability concerns underlying the current semisynthetic process encouraged us to immobilize the engineered acyltransferase LovD−BuCh2 on different pore carriers to develop an innovative and sustainable process for the continuous biomanufacturing of simvastatin. We systematically assessed the effect of the immobilization on the functional and structural properties of the immobilized enzyme, the enzyme spatial distribution across the solid material, and the thermal stability of the immobilized biocatalysts. After screening several immobilization carriers, we selected LovD−BuCh2 immobilized on the controlled porous glass particles functionalized with Fe 3+ − catechol complexes as the most suitable heterogeneous biocatalyst to optimize the continuous synthesis of simvastatin. This immobilized enzyme was four times more thermally stable than its free counterpart and maintained >60% product yield during five operational cycles in the batch. Under the optimal flow conditions, we achieved 100% of simvastatin yield with a space-time yield of 4.61 g L −1 h −1 and a specific productivity of 17 mg product × mg enzyme −1 L −1 . This flow-biocatalysis system improves some green chemistry metrics such as the reaction mass efficiency and the atom economy when compared to previous studies of simvastatin manufacturing.

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Section: Resultsmentioning

confidence: 98%

Immobilization and Stabilization of an Engineered Acyltransferase for the Continuous Biosynthesis of Simvastatin in Packed-Bed Reactors

García‐Marquina

Langer

Sánchez-Costa

et al. 2022

ACS Sustainable Chem. Eng.

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“…One advantage of enzymatic catalysis in an organic solvent system is that the enzyme can be effectively immobilised by simple adsorption onto an inert support, which provides a large surface area and high protein loading capacity (Ampon 1992). Immobilised lipases were produced with 45.8% (103.05 mg protein g (1 support) and 71.9% (161.75 mg protein g (1 support) protein loading for Ni-LDH and Ni-SDS-LDH nanocomposites, respectively (Table II).…”

Section: Immobilisation Of Lipasementioning

confidence: 99%

Application of advanced materials as support for immobilisation of lipase fromCandida rugosa

Rahman

Yunus

Hussein

et al. 2005

Biocatalysis and Biotransformation

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Ni/Al-layered double hydroxides (Ni-LDHs) and Ni/Al-sodium dodecyl sulfonate layered double hydroxide nanocomposites (Ni-SDS-LDHs) with a molar ratio of Ni:Al (4:1) have been prepared by a co-precipitation (or salt-base) method. Their structures were determined using Powder X-Ray Diffractometer (PXRD) and the spectra showed that basal spacings for Ni-LDHs and Ni-SDS-LDHs synthesised were around 8.1 Å and 34.8 Å , respectively. Lipase from Candida rugosa was immobilised onto these advanced materials, by physical adsorption. The activity of immobilised lipase was investigated through esterification of palmitic acid and isopropyl alcohol in hexane. The effects of reaction temperature, thermostability, stability in organic solvent, operational stability, leaching and storage studies of the immobilised lipase were investigated. These biocatalysts exhibited higher activities than the native lipase with an optimum temperature of 408C. Immobilised lipases showed higher storage stability than native lipase (up to 60 days) and during operational studies at 308C for 5 h, more than 50% of its activity was retained. Leaching studies showed that physical adsorption is suitable for the attachment of enzymes onto LDHs.

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“…On the other hand, we also wanted to quantitatively measure the enzyme distribution in the immobilized derivatives in order to rationally design new improved biocatalysts. Staining with different colored dyes (such as Ponceau S and naphthol blue black) has been used to study enzyme distribution in immobilized biocatalysts (Carleysmith et al 1980;Ampon 1992). Other methods require tagging of the enzyme (e.g.…”

Section: Introductionmentioning

confidence: 99%

Kinetic and microstructural characterization of immobilized penicillin acylase fromStreptomyces lavendulaeon Sepabeads EC-EP

Hormigo

Mata

Castillón

et al. 2009

Biocatalysis and Biotransformation

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Recombinant penicillin acylase from Streptomyces lavendulae was covalently bound to epoxy-activated Sepabeads EC-EP303 † . Optimization of the immobilization process led to a homogeneous distribution of the enzyme on the support surface avoiding the attachment of enzyme aggregates, as shown by confocal electron microscopy. The optimal immobilized biocatalyst had a specific enzymatic activity of 26.2 IU g wet carrier (1 in the hydrolysis of penicillin V at pH 8.0 and 408C. This biocatalyst showed the highest activity at pH 8.5 and 658C, 1.5 pH units lower and 58C higher than its soluble counterpart. Substrate specificity of the derivative also showed its ability to efficiently hydrolyze other natural aliphatic penicillins such as penicillins K, F and dihydroF. The immobilized enzyme was highly stable at 408C and pH 8.0 (t 1/2 0625 h vs. t 1/2 0397 h for the soluble enzyme), and it could be recycled for at least 30 consecutive batch reactions without loss of catalytic activity.

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Distribution of an enzyme in porous polymer beads

Cited by 11 publications

References 24 publications

Immobilization and Stabilization of an Engineered Acyltransferase for the Continuous Biosynthesis of Simvastatin in Packed-Bed Reactors

Immobilization and Stabilization of an Engineered Acyltransferase for the Continuous Biosynthesis of Simvastatin in Packed-Bed Reactors

Application of advanced materials as support for immobilisation of lipase fromCandida rugosa

Kinetic and microstructural characterization of immobilized penicillin acylase fromStreptomyces lavendulaeon Sepabeads EC-EP

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