scholarly journals “Fishing and Hunting”—Selective Immobilization of a Recombinant Phenylalanine Ammonia-Lyase from Fermentation Media

Molecules ◽  
2019 ◽  
Vol 24 (22) ◽  
pp. 4146 ◽  
Author(s):  
Sánta-Bell ◽  
Molnár ◽  
Varga ◽  
Nagy ◽  
Hornyánszky ◽  
...  
Keyword(s):  
Metal Ion ◽  
Enzyme Purification ◽  
Phenylalanine Ammonia Lyase ◽  
Covalent Bond ◽  
Affinity Tag ◽  
Whole Cells ◽  
Bond Forming ◽  
Downstream Processes ◽  
Selective Immobilization ◽  
Cell Fragments

This article overviews the numerous immobilization methods available for various biocatalysts such as whole-cells, cell fragments, lysates or enzymes which do not require preliminary enzyme purification and introduces an advanced approach avoiding the costly and time consuming downstream processes required by immobilization of purified enzyme-based biocatalysts (such as enzyme purification by chromatographic methods and dialysis). Our approach is based on silica shell coated magnetic nanoparticles as solid carriers decorated with mixed functions having either coordinative binding ability (a metal ion complexed by a chelator anchored to the surface) or covalent bond-forming ability (an epoxide attached to the surface via a proper linker) enabling a single operation enrichment and immobilization of a recombinant phenylalanine ammonia-lyase from parsley fused to a polyhistidine affinity tag.

scholarly journals Immobilization of Phenylalanine Ammonia-lyase via EDTA Based Metal Chelate Complexes – Optimization and Prospects

2021 ◽  
Author(s):  
Evelin Sánta-Bell ◽  
Norbert Krisztián Kovács ◽  
Bálint Alács ◽  
Zsófia Molnár ◽  
Gábor Hornyánszky
Keyword(s):  
Metal Ions ◽  
Metal Ion ◽  
Phenylalanine Ammonia Lyase ◽  
Metal Chelate ◽  
Maximal Activity ◽  
Transition Metal Ions ◽  
Enzyme Inactivation ◽  
Polymeric Carrier ◽  
Activity Yield ◽  
Selective Immobilization

Immobilized metal ion affinity chromatography principles were applied for selective immobilization of recombinant polyhistidine tag fused phenylalanine ammonia-lyase from parsley (PcPAL) on porous polymeric support with aminoalkyl moieties modified with an EDTA dianhydride (EDTADa)-derived chelator and charged with transition metal ions. Out of the five investigated metal ions - Fe3+, Co2+, Ni2+, Cu2+, Zn2+ - the best biocatalytic activity of PcPAL was achieved when the enzyme was immobilized on the Co2+ ion-charged support (31.8 ± 1.2 U/g). To explore the features this PcPAL obtained by selective immobilization, the thermostability and reusability of this PAL biocatalyst were investigated. To maximize the activity of the immobilized PcPAL the surface functionalization of the aminoalkylated polymeric carrier was fine-tuned with using glycidol as a thinning group beside EDTADa. The maximal activity yield (YA=103 %) was earned when the EDTADa and glycidol were used in 1 to 24 ratio. The reversibility of the immobilization method allowed the development of a support regeneration protocol which enables easy reuse of the functionalized support in case of enzyme inactivation.

Rietveld refinement of the cocrystal 2,4-dihydroxybenzoic acid–N′-(propan-2-ylidene)nicotinohydrazide (1/1)

2012 ◽  
Vol 68 (9) ◽  
pp. o335-o337 ◽  
Author(s):  
Saul H. Lapidus ◽  
Andreas Lemmerer ◽  
Joel Bernstein ◽  
Peter W. Stephens
Keyword(s):  
Hydrogen Bond ◽  
Hydrogen Bonding ◽  
Powder Diffraction ◽  
Supramolecular Assembly ◽  
Covalent Bond ◽  
Analytical Tool ◽  
Powder Diffraction Data ◽  
Dihydroxybenzoic Acid ◽  
Bond Forming ◽  
Hydrogen Bond Interactions

A further example of using a covalent-bond-forming reaction to alter supramolecular assembly by modification of hydrogen-bonding possibilities is presented. This concept was introduced by Lemmerer, Bernstein & Kahlenberg [CrystEngComm(2011),13, 55–59]. The title structure, C9H11N3O·C7H6O4, which consists of a reacted niazid molecule,viz.N′-(propan-2-ylidene)nicotinohydrazide, and 2,4-dihydroxybenzoic acid, was solved from powder diffraction data using simulated annealing. The results further demonstrate the relevance and utility of powder diffraction as an analytical tool in the study of cocrystals and their hydrogen-bond interactions.

Theoretical Study of Iron Heterogeneous Growth on the Surface of C60 Molecule

2014 ◽  
Vol 1081 ◽  
pp. 115-118
Author(s):  
Qi Cheng Liu ◽  
Yun Fang Li
Keyword(s):  
Crystallization Process ◽  
Theoretical Study ◽  
Covalent Bond ◽  
Md Simulations ◽  
Nucleation And Growth ◽  
Bond Forming ◽  
Regular Icosahedron ◽  
Molecule Dynamics ◽  
Molecule Surface ◽  
Regular Closed

Direct molecule dynamics (MD) simulations have also been performed to study heterogeneous nucleation and growth of iron on C60 molecule. The grown mechanism of this crystallization process was explored. The results indicate that 92 iron atoms attach to C60 molecule surface can form new covalent bond, forming a closed regular icosahedron. More atoms grow in layer to form bigger regular closed clathrate base on the structure of former one. As increase of atoms number, there will appear some crystal faces.

The impact of species, respiration type, growth phase and genetic inventory on absolute metal content of intact bacterial cells

Metallomics ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 925-935 ◽  
Author(s):  
Rohit Budhraja ◽  
Chang Ding ◽  
Philipp Walter ◽  
Stephan Wagner ◽  
Thorsten Reemtsma ◽  
...  
Keyword(s):  
Growth Phase ◽  
Metal Ion ◽  
Metal Content ◽  
Growth Conditions ◽  
Bacterial Cells ◽  
Ion Content ◽  
Whole Cells ◽  
Microbial Species ◽  
The Impact

Absolute metal ion content was determined from whole cells of different microbial species and changes were related to growth conditions and change of encoded genes.

A quantum mechanical approach to ligand binding — Calculation of ligand–protein binding affinities for stromelysin-1 (MMP-3) inhibitors

2009 ◽  
Vol 87 (10) ◽  
pp. 1480-1484 ◽  
Author(s):  
Jian Li ◽  
Charles. H. Reynolds
Keyword(s):  
Ligand Binding ◽  
Metal Ion ◽  
Protein Complexes ◽  
Field Method ◽  
Binding Energies ◽  
Quantum Mechanical ◽  
Binding Affinities ◽  
Bond Forming ◽  
Proton Charge ◽  
Quantum Mechanical Approach

Linear-scaling quantum mechanical method was applied to calculate binding affinities of six stromelysin-1 (MMP-3) inhibitors with two different zinc binding groups (ZBGs). The entire protein and ligand–protein complexes were calculated using PM5 Hamiltonian, which enables the treatment of metal ion coordination, bond forming/breaking, and proton/charge transfers associated with the ligand binding process by the self-consistent field method. The calculated binding energies reproduce the binding-affinity trend observed experimentally.

scholarly journals Supercontracted state of vertebrate smooth muscle cell fragments reveals myofilament lengths.

1990 ◽  
Vol 111 (6) ◽  
pp. 2451-2461 ◽  
Author(s):  
J V Small ◽  
M Herzog ◽  
M Barth ◽  
A Draeger
Keyword(s):  
Smooth Muscle ◽  
Actin Filaments ◽  
Cytoskeletal Proteins ◽  
Terminal Phase ◽  
Myosin Filament ◽  
Protein Loss ◽  
Whole Cells ◽  
Myosin Filaments ◽  
Central Bundle ◽  
Cell Fragments

Isolated cell preparations from chicken gizzard smooth muscle typically contain a mixture of cell fragments and whole cells. Both species are spontaneously permeable and may be preloaded with externally applied phalloidin and antibodies and then induced to contract with Mg ATP. Labeling with antibodies revealed that the cell fragments specifically lacked certain cytoskeletal proteins (vinculin, filamin) and were depleted to various degrees in others (desmin, alpha-actinin). The cell fragments showed a unique mode of supercontraction that involved the protrusion of actin filaments through the cell surface during the terminal phase of shortening. In the presence of dextran, to minimize protein loss, the supercontracted products were star-like in form, comprising long actin bundles radiating in all directions from a central core containing myosin, desmin, and alpha-actinin. It is concluded that supercontraction is facilitated by an effective uncoupling of the contractile apparatus from the cytoskeleton, due to partial degradation of the latter, which allows unhindered sliding of actin over myosin. Homogenization of the cell fragments before or after supercontraction produced linear bipolar dimer structures composed of two oppositely polarized bundles of actin flanking a central bundle of myosin filaments. Actin filaments were shown to extend the whole length of the bundles and their length averaged integral to 4.5 microns. Myosin filaments in the supercontracted dimers averaged 1.6 microns in length. The results, showing for the first time the high actin to myosin filament length ratio in smooth muscle are readily consistent with the slow speed of shortening of this tissue. Other implications of the results are also discussed.

scholarly journals HNF4α Is a Covalent Bond-Forming Receptor

2021 ◽  
Vol 67 (2) ◽  
pp. 126-129
Author(s):  
Satoshi KANAMORI ◽  
Nami OHASHI ◽  
Hiroaki ISHIDA ◽  
Keiko YAMAMOTO ◽  
Toshimasa ITOH
Keyword(s):  
Covalent Bond ◽  
Bond Forming
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