Co-immobilised aspartase and transaminase for high-yield synthesis of l-phenylalanine

Cárdenas-Fernández, Max; Khalikova, Elvira; Korpela, Timo; López, Carmen; Álvaro, Gregorio

doi:10.1016/j.bej.2014.10.010

Cited by 12 publications

(8 citation statements)

References 23 publications

(35 reference statements)

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“…One example is the recent co-immobilisation of AspB with a microbial aspartate transaminase (AspTA) to produce L-phenylalanine from phenylpyruvate, using as the amino donor L-aspartate formed in situ from fumarate (Scheme 17a). 203 The two enzymes were co-immobilized on an amino-epoxy support, giving a 95% yield of L-phenylalanine at 100 mM concentration in continuous flow. In another example, Zhu and co-workers employed aspartase as an ammonia scavenger to shift the equilibria of a nitrilase reaction for the production of β-alanine (Scheme 17b).…”

Section: Synthesis Of Aspartic Acid Derivativesmentioning

confidence: 99%

Synthetic and Therapeutic Applications of Ammonia-lyases and Aminomutases

et al. 2017

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Ammonia-lyases and aminomutases are mechanistically and structurally diverse enzymes which catalyze the deamination and/or isomerization of amino acids in nature by cleaving or shifting a C-N bond. Of the many protein families in which these enzyme activities are found, only a subset have been employed in the synthesis of optically pure fine chemicals or in medical applications. This review covers the natural diversity of these enzymes, highlighting particular enzyme classes that are used within industrial and medical biotechnology. These highlights detail the discovery and mechanistic investigations of these commercially relevant enzymes, along with comparisons of their various applications as stand-alone catalysts, components of artificial biosynthetic pathways and biocatalytic or chemoenzymatic cascades, and therapeutic tools for the potential treatment of various pathologies.

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Section: Synthesis Of Aspartic Acid Derivativesmentioning

confidence: 99%

Synthetic and Therapeutic Applications of Ammonia-lyases and Aminomutases

et al. 2017

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“…[3] Furthermore, AAL has been coimmobilized with a transaminase on amino-epoxy ReliZyme® support and used for the production of l-phenylalanine. [25] Magnetic nanoparticles (MNPs) attracted attention in various fields of biotechnology as suitable carriers for enzyme immobilization. [26,27] MNPs have several advantageous features such as high surface area and low mass transfer resistanceresulting in high reactivity-, and simple isolation of the immobilized biocatalyst from the reaction mixture.…”

Section: Introductionmentioning

confidence: 99%

“…AALs were immobilized on epoxy (Eupergit® C) and amine (MANA‐agarose) supports, [3] or entrapped in polyacrylamide gel [24] and polyvinyl alcohol (LentiKats®) [3] . Furthermore, AAL has been co‐immobilized with a transaminase on amino‐epoxy ReliZyme® support and used for the production of l ‐phenylalanine [25] …”

Section: Introductionmentioning

confidence: 99%

Immobilization of the Aspartate Ammonia‐Lyase from Pseudomonas fluorescens R124 on Magnetic Nanoparticles: Characterization and Kinetics

et al. 2022

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Aspartate ammonia-lyases (AALs) catalyze the non-oxidative elimination of ammonia from l-aspartate to give fumarate and ammonia. In this work the AAL coding gene from Pseudomonas fluorescens R124 was identified, isolated, and cloned into the pET-15b expression vector and expressed in E. coli. The purified enzyme (PfAAL) showed optimal activity at pH 8.8, Michaelis-Menten kinetics in the ammonia elimination from l-aspartate, and no strong dependence on divalent metal ions for its activity. The purified PfAAL was covalently immobilized on epoxy-functionalized magnetic nanoparticles (MNP), and effective kinetics of the immobilized PfAAL-MNP was compared to the native solution form. Glycerol addition significantly enhanced the storability of PfAAL-MNP. Inhibiting effect of the growing viscosity (modulated by addition of glycerol or glucose) on the enzymatic activity was observed for the native and immobilized form of PfAAL, as previously described for other free enzymes. The storage stability and recyclability of PfAAL-MNP is promising for further biocatalytic applications.

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“…Genetically, engineered transaminases have demonstrated excellent performance in the synthesis of important drug scaffolds [5]. In addition, keeping the recyclability issue in mind, researchers have proposed successful immobilization strategies for transaminases using for example natural and synthetic polymers [8][9][10][11][12] or silica [13][14][15] as carriers.…”

Section: Introductionmentioning

confidence: 99%

Enantioselective Transamination in Continuous Flow Mode with Transaminase Immobilized in a Macrocellular Silica Monolith

2017

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ω-Transaminases have been immobilized on macrocellular silica monoliths and used as heterogeneous biocatalysts in a continuous flow mode enantioselective transamination reaction. The support was prepared by a sol-gel method based on emulsion templating. The enzyme was immobilized on the structured silica monoliths both by adsorption, and by covalent grafting using amino-functionalized silica monoliths and glutaraldehyde as a coupling agent. A simple reactor set-up based on the use of a heat-shrinkable Teflon tube is presented and successfully used for the continuous flow kinetic resolution of a chiral amine, 4-bromo-α-methylbenzylamine. The porous structure of the supports ensures effective mass transfer and the reactor works in the plug flow regime without preferential flow paths. When immobilized in the monolith and used in the flow reactor, transaminases retain their activity and their enantioselectivity. The solid biocatalyst is also shown to be stable both on stream and during storage. These essential features pave the way to the successful development of an environmentally friendly process for chiral amines production.

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Co-immobilised aspartase and transaminase for high-yield synthesis of l-phenylalanine

Cited by 12 publications

References 23 publications

Synthetic and Therapeutic Applications of Ammonia-lyases and Aminomutases

Synthetic and Therapeutic Applications of Ammonia-lyases and Aminomutases

Immobilization of the Aspartate Ammonia‐Lyase from Pseudomonas fluorescens R124 on Magnetic Nanoparticles: Characterization and Kinetics

Enantioselective Transamination in Continuous Flow Mode with Transaminase Immobilized in a Macrocellular Silica Monolith

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