Recent developments in biocatalysis beyond the laboratory

Narancic, Tanja; Davis, Reeta; Nikodinovic-Runic, Jasmina; Connor, Kevin E. O’

doi:10.1007/s10529-014-1762-4

Cited by 53 publications

(34 citation statements)

References 72 publications

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“…We describe an improved strategy for the generation of tailored core-shell related nanoparticles with several genetically encoded and tunable functionalities including magnetism, the presence of a biological envelope, fluorescence, and catalytic activity. Since the stable expression of catalytically active enzyme proteins, their immobilization, and reusability are still common challenges, [50] this provides a highly promising route also for the display of other enzyme proteins and peptides more relevant Table 3. Protein amounts, copy numbers, and enzymatic activities of magnetosomes displaying GusA arrays on the particle surface.…”

Section: Resultsmentioning

confidence: 99%

Generation of Multifunctional Magnetic Nanoparticles with Amplified Catalytic Activities by Genetic Expression of Enzyme Arrays on Bacterial Magnetosomes

Mickoleit

Schüler

2017

Adv. Biosys.

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Due to their highly regulated biosynthesis, magnetosomes biomineralized by magnetotactic bacteria represent natural magnetic nanoparticles with unique physical and chemical properties. They consist of a magnetite core that is surrounded by a biological membrane and are therefore reminiscent to magnetic “core–shell” nanoparticles. Their usability in many nanotechnological and biomedical applications would be further improved by the introduction of additional catalytic and imaging modalities. Here, a new in vivo strategy is explored for magnetosome display of foreign polypeptides with maximized protein‐to‐particle ratios. Arrays of up to five monomers of the model enzyme glucuronidase GusA plus the additional fluorophore mEGFP are genetically fused as single large hybrid proteins to highly expressed magnetosome protein anchors. In total, about 190 GusA monomers are covalently attached to individual particles. Assuming layers of GusA rows surrounding the particles, the monomers would thus cover up to 90% of the magnetosome surface. The approach generates nanoparticles that exhibit magnetism, fluorescence, and stable catalytic activities, which are stepwise increased with the number of GusA monomers. In summary, multicopy expression of arrayed foreign proteins represents a powerful methodology for the biosynthesis of tailored biohybrid magnetic nanoparticles with several genetically encoded and tunable functionalities.

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Section: Resultsmentioning

confidence: 99%

Generation of Multifunctional Magnetic Nanoparticles with Amplified Catalytic Activities by Genetic Expression of Enzyme Arrays on Bacterial Magnetosomes

Mickoleit

Schüler

2017

Adv. Biosys.

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“…Biocatalytic processes are increasingly being considered in industrial organic syntheses since in certain situations they are considered 'greener' and can bring considerable reductions to overall process costs [1,2]. This is a consequence of the mild temperature, pressure and pH conditions under which enzymes function.…”

Section: Introductionmentioning

confidence: 99%

High throughput screening of monoamine oxidase (MAO-N-D5) substrate selectivity and rapid kinetic model generation

Rios‐Solis

Mothia

et al. 2015

Journal of Molecular Catalysis B: Enzymatic

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a b s t r a c tFull kinetic models provide insight into enzyme mechanism and kinetics and also support bioconversion process design and feasibility assessment. Previously we have established automated microwell methods for rapid data collection and hybrid kinetic modelling techniques for quantification of kinetic constants. In this work these methods are applied to explore the substrate selectivity and kinetics of monoamine oxidase, MAO-N-D5, from Aspergillus niger. In particular we examine the MAO-N-D5 variant Ile246Met/Asn336Ser/Met348Lys/Thr384Asn to allow the oxidation of secondary amines Initial screening showed that MAO-N-D5 enabled the selective oxidation of secondary amines in 8 and 9 carbon rings, as well as primary ethyl and propyl amines attached to secondary amines of indolines and pyrrolidines. Subsequently we developed a first kinetic model for the MAO-N-D5 enzyme based on the ping-pong bi-bi mechanism (similar to that for the human MAO-A enzyme). The full set of kinetic parameters were then established for three MAO-N-D5 substrates namely; 3-azabicyclo[3,3,0]octane, 1-(2 amino ethyl) pyrrolidine and 3-(2,3-dihydro-1H-indole-1-yl)propan-1-amine. The models for each amine substrate showed excellent agreement with experimentally determined progress curves over a range of operating conditions. They indicated that in each case amine inhibition was the main determinant of overall reaction rate rather than oxygen or imine (product) inhibition. From the perspective of larger scale bioconversion process design, the models indicated the need for fed-batch addition of the amine substrate and to increase the dissolved oxygen levels in order to maximize bioconversion process productivity.

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“…Moreover, lipases do not usually require cofactors, are quite robust and stable, and can be active in organic solvents [1,18–20]. However, not all lipolytic enzymes bear the properties required for a defined biotechnological process, and availability of new catalysts with striking properties is still a challenge [21,22]. For those reactions requiring harsh conditions like high temperature or extreme pH, different strategies can be followed.…”

Section: Introductionmentioning

confidence: 99%

Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past

et al. 2017

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A search for extremophile enzymes from ancient volcanic soils in El Hierro Island (Canary Islands, Spain) allowed isolation of a microbial sporulated strain collection from which several enzymatic activities were tested. Isolates were obtained after sample cultivation under several conditions of nutrient contents and temperature. Among the bacterial isolates, supernatants from the strain designated JR3 displayed high esterase activity at temperatures ranging from 30 to 100°C, suggesting the presence of at least a hyper-thermophilic extracellular lipase. Sequence alignment of known thermophilic lipases allowed design of degenerated consensus primers for amplification and cloning of the corresponding lipase, named LipJ. However, the cloned enzyme displayed maximum activity at 30°C and pH 7, showing a different profile from that observed in supernatants of the parental strain. Sequence analysis of the cloned protein showed a pentapeptide motif -GHSMG- distinct from that of thermophilic lipases, and much closer to that of esterases. Nevertheless, the 3D structural model of LipJ displayed the same folding as that of thermophilic lipases, suggesting a common evolutionary origin. A phylogenetic study confirmed this possibility, positioning LipJ as a new member of the thermophilic family of bacterial lipases I.5. However, LipJ clusters in a clade close but separated from that of Geobacillus sp. thermophilic lipases. Comprehensive analysis of the cloned enzyme suggests a common origin of LipJ and other bacterial thermophilic lipases, and highlights the most probable divergent evolutionary pathway followed by LipJ, which during the harsh past times would have probably been a thermophilic enzyme, having lost these properties when the environment changed to more benign conditions.

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Recent developments in biocatalysis beyond the laboratory

Cited by 53 publications

References 72 publications

Generation of Multifunctional Magnetic Nanoparticles with Amplified Catalytic Activities by Genetic Expression of Enzyme Arrays on Bacterial Magnetosomes

Generation of Multifunctional Magnetic Nanoparticles with Amplified Catalytic Activities by Genetic Expression of Enzyme Arrays on Bacterial Magnetosomes

High throughput screening of monoamine oxidase (MAO-N-D5) substrate selectivity and rapid kinetic model generation

Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past

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