“I de 11/2 år jeg hat arbejdet på AGC biologics. Her Keith været min teamlader. Det har været en sand fornæjelse at have Keith som daglige leder. Han er super kompentent til at jonglere alle bolde i luften, som høre med til et teams leders arbejdsbord. Keith er en meget glad og smillende person. Han er meget hjælpsom og flittig. Han er der altid dagligt for sit team, også i situantioner som er svære at klare. Han er som en blæksprutte der jonglere sit talent mellem 2 teams og det klare han til UG.”
København, Region Hovedstaden, Danmark
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Happy 5th Anniversary, Solar Biotech! 🎉 Today, we kick off our celebrations as we mark 5 years of remarkable achievements and growth. 🌟 Reflecting…
Happy 5th Anniversary, Solar Biotech! 🎉 Today, we kick off our celebrations as we mark 5 years of remarkable achievements and growth. 🌟 Reflecting…
Keith McCall synes godt om dette
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We have an open Scientist position with my team in NORCE's Industrial Biotechnology group. The team is doing cutting edge work in both precision…
We have an open Scientist position with my team in NORCE's Industrial Biotechnology group. The team is doing cutting edge work in both precision…
Delt af Keith McCall
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Proud to be part of AGC Biologics💚
Proud to be part of AGC Biologics💚
Keith McCall synes godt om dette
Erfaring og uddannelse
Erfaring med frivilligt arbejde
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Mentor
IndieBio
– nu 9 år 7 måneder
Videnskab og teknologi
Mentoring small biotech startups in a biotech incubator.
Udgivelser
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Metabolic engineering of Aspergillus oryzae NRRL 3488 for increased production of L-malic acid
Applied Microbiology and Biotechnology
Malic acid, a petroleum-derived C4-dicarboxylic acid that is used in the food and beverage industries, is also produced by a number of microorganisms that follow a variety of metabolic routes. Several members of the genus Aspergillus utilize a two-step cytosolic pathway from pyruvate to malate known as the reductive tricarboxylic acid (rTCA) pathway. This simple and efficient pathway has a maximum theoretical yield of 2 mol malate/mol glucose when the starting pyruvate originates from…
Malic acid, a petroleum-derived C4-dicarboxylic acid that is used in the food and beverage industries, is also produced by a number of microorganisms that follow a variety of metabolic routes. Several members of the genus Aspergillus utilize a two-step cytosolic pathway from pyruvate to malate known as the reductive tricarboxylic acid (rTCA) pathway. This simple and efficient pathway has a maximum theoretical yield of 2 mol malate/mol glucose when the starting pyruvate originates from glycolysis. Production of malic acid by Aspergillus oryzae NRRL 3488 was first improved by overexpression of a native C4-dicarboxylate transporter, leading to a greater than twofold increase in the rate of malate production. Overexpression of the native cytosolic alleles of pyruvate carboxylase and malate dehydrogenase, comprising the rTCA pathway, in conjunction with the transporter resulted in an additional 27 % increase in malate production rate. A strain overexpressing all three genes achieved a malate titer of 154 g/L in 164 h, corresponding to a production rate of 0.94 g/L/h, with an associated yield on glucose of 1.38 mol/mol (69 % of the theoretical maximum). This rate of malate production is the highest reported for any microbial system.
Andre forfattereSe udgivelse -
Zap1 activation domain 1 and its role in controlling gene expression in response to cellular zinc status
Molecular Microbiology
The Zap1 transcription factor is a central player in zinc homeostasis in yeast. This protein regulates the expression of genes involved in zinc accumulation and storage. For most of its target genes, Zap1 activates expression in zinc-limited cells and this function is inhibited in replete cells. Zap1 has two activation domains, AD1 and AD2, which are independently regulated by zinc status. In this study, we characterized AD1 and its regulation by zinc. AD1 was mapped using deletions to residues…
The Zap1 transcription factor is a central player in zinc homeostasis in yeast. This protein regulates the expression of genes involved in zinc accumulation and storage. For most of its target genes, Zap1 activates expression in zinc-limited cells and this function is inhibited in replete cells. Zap1 has two activation domains, AD1 and AD2, which are independently regulated by zinc status. In this study, we characterized AD1 and its regulation by zinc. AD1 was mapped using deletions to residues 332-402 of Zap1. The region required for the zinc responsiveness of this activation domain, designated 'ZRD(AD1), was mapped to residues 182-502. Thus, AD1 is embedded within its larger zinc-responsive domain. Using a combination of in silico analysis, random mutagenesis and site-directed mutagenesis, we identified key residues within ZRD(AD1) required for its regulation by zinc. Most of these residues are cysteines and histidines that could potentially serve as Zn(II) ligands. These results suggest that ZRD(AD1) senses zinc by direct Zn(II) binding. Consistent with this hypothesis, purified ZRD(AD1) bound multiple Zn(II) ions. Finally, our results indicate that, in the context of the full-length Zap1 protein, AD1 and AD2 are both critical to the full control of gene expression in response to zinc.
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Probing determinants of the metal ion selectivity in carbonic anhydrase using mutagenesis
Biochemistry
Using Carbonic Anhydrase as a model enzyme, various hypotheses regarding metal ion selectivity of metalloproteins were tested through site-directed mutagenesis and binding kinetics studies.
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Zinc fingers can act as Zn2+ sensors to regulate transcriptional activation domain function.
EMBO J.
An investigation into the zinc mediation of the yeast Zap1 transcription factor.
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Chapter 19. Mitochondrial Copper Ion Transport, in Microbial Transport Systems (ed G. Winkelmann)
Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, FRG
This chapter contains sections titled:
Introduction
Mitochondrial Structure
Mitochondrial Transport
Assembly of Mitochondrial Cytochrome c Oxidase
Copper Ion Delivery to Targets other than the Mitochondrion
Copper Ion Transport to the Mitochondrion by Cox17
Co-metallochaperones in Cu Metallation of Cytochrome c Oxidase
Terminal Oxidases in Prokaryotes
Metallation of Prokaryotic Terminal Oxidases
Postulated Model -
Thermodynamics of metal ion binding. 2. Metal ion binding by carbonic anhydrase variants.
Biochemistry
The ability to construct molecular motifs with predictable properties in aqueous solution requires an extensive knowledge of the relationships between structure and energetics. The design of metal binding motifs is currently an area of intense interest in the bioorganic community. To date synthetic motifs designed to bind metal ions lack the remarkable affinities observed in biological systems. To better understand the structural basis of metal ion affinity, we report here the thermodynamics of…
The ability to construct molecular motifs with predictable properties in aqueous solution requires an extensive knowledge of the relationships between structure and energetics. The design of metal binding motifs is currently an area of intense interest in the bioorganic community. To date synthetic motifs designed to bind metal ions lack the remarkable affinities observed in biological systems. To better understand the structural basis of metal ion affinity, we report here the thermodynamics of binding of divalent zinc ions to wild-type and mutant carbonic anhydrases and the interpretation of these parameters in terms of structure. Mutations were made both to the direct His ligand at position 94 and to indirect, or second-shell, ligands Gln-92, Glu-117, and Thr-199. The thermodynamics of ligand binding by several mutant proteins is complicated by the development of a second zinc binding site on mutation; such effects must be considered carefully in the interpretation of thermodynamic data. In all instances modification of the protein produces a complex series of changes in both the enthalpy and entropy of ligand binding. In most cases these effects are most readily rationalized in terms of ligand and protein desolvation, rather than in terms of changes in the direct interactions of ligand and protein. Alteration of second-shell ligands, thought to function primarily by orienting the direct ligands, produces profoundly different effects on the enthalpy of binding, depending on the nature of the residue. These results suggest a range of activities for these ligands, contributing both enthalpic and entropic effects to the overall thermodynamics of binding. Together, our results demonstrate the importance of understanding relationships between structure and hydration in the construction of novel ligands and biological polymers.
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Thermodynamics of Metal Ion Binding. 1. Metal Ion Binding by Wild-Type Carbonic Anhydrase
Biochemistry
Understanding the energetic consequences of molecular structure in aqueous solution is a prerequisite to the rational design of synthetic motifs with predictable properties. Such properties include ligand binding and the collapse of polymer chains into discrete three-dimensional structures. Despite advances in macromolecular structure determination, correlations of structure with high-resolution thermodynamic data remain limited. Here we compare thermodynamic parameters for the binding of…
Understanding the energetic consequences of molecular structure in aqueous solution is a prerequisite to the rational design of synthetic motifs with predictable properties. Such properties include ligand binding and the collapse of polymer chains into discrete three-dimensional structures. Despite advances in macromolecular structure determination, correlations of structure with high-resolution thermodynamic data remain limited. Here we compare thermodynamic parameters for the binding of Zn(II), Cu(II), and Co(II) to human carbonic anhydrase II. These calorimetrically determined values are interpreted in terms of high-resolution X-ray crystallographic data. While both zinc and cobalt are bound with a 1:1 stoichiometry, CAII binds two copper ions. Considering only the high-affinity site, there is a diminution in the enthalpy of binding through the series Co(II) → Zn(II) → Cu(II) that mirrors the enthalpy of hydration; this observation reinforces the notion that the thermodynamics of solute association with water is at least as important as the thermodynamics of solute−solute interaction and that these effects must be considered when interpreting association in aqueous solution. Additionally, ΔCp data suggest that zinc binding to CAII proceeds with a greater contribution from desolvation than does binding of either copper or cobalt, suggesting Nature optimizes binding by optimizing desolvation.
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Colorimetric and fluorimetric assays to quantitate micromolar concentrations of transition metals
Analytical Biochemistry
Transition metal ions, although maintained at low concentrations, play diverse important roles in many biological processes. Two assays useful for the rapid quantification of a range of first-row transition metal ions have been developed. The colorimetric assay extends the 4-(2-pyridylazo)resorcinol assay of Hunt et al. (J. Biol. Chem. 255, 14793 (1984)) to measure nanomole quantities of Co2+, Ni2+, and Cu2+ as well as Zn2+. The fluorimetric assay takes advantage of the coordination of a number…
Transition metal ions, although maintained at low concentrations, play diverse important roles in many biological processes. Two assays useful for the rapid quantification of a range of first-row transition metal ions have been developed. The colorimetric assay extends the 4-(2-pyridylazo)resorcinol assay of Hunt et al. (J. Biol. Chem. 255, 14793 (1984)) to measure nanomole quantities of Co2+, Ni2+, and Cu2+ as well as Zn2+. The fluorimetric assay takes advantage of the coordination of a number of metal ions (Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+) by Fura-2 and can also be used to measure nanomole quantities of these ions. The assays developed here have the advantage of not requiring the extensive sample preparation necessary for other methodologies, such as atomic absorption spectroscopy and inductively coupled plasma emission spectroscopy (ICPES), while being comparable in accuracy to the detection limits of ICPES for the first-row transition metal ions. To demonstrate the effectiveness of these assays, we determined the affinity of carbonic anhydrase II (CA II), a prototypical zinc enzyme, for Ni2+ and Cd2+. These data indicate that CA II binds transition metals with high affinity and is much more selective for Zn2+ over Ni2+ or Cd2+ than most small-molecule chelators or other metalloenzymes.
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Function and mechanism of zinc metalloenzymes
The Journal of Nutrition
Zinc is required for the activity of > 300 enzymes, covering all six classes of enzymes. Zinc binding sites in proteins are often distorted tetrahedral or trigonal bipyramidal geometry, made up of the sulfur of cysteine, the nitrogen of histidine or the oxygen of aspartate and glutamate, or a combination. Zinc in proteins can either participate directly in chemical catalysis or be important for maintaining protein structure and stability. In all catalytic sites, the zinc ion functions as a…
Zinc is required for the activity of > 300 enzymes, covering all six classes of enzymes. Zinc binding sites in proteins are often distorted tetrahedral or trigonal bipyramidal geometry, made up of the sulfur of cysteine, the nitrogen of histidine or the oxygen of aspartate and glutamate, or a combination. Zinc in proteins can either participate directly in chemical catalysis or be important for maintaining protein structure and stability. In all catalytic sites, the zinc ion functions as a Lewis acid. Researchers in our laboratory are dissecting the determinants of molecular recognition and catalysis in the zinc-binding site of carbonic anhydrase. These studies demonstrate that the chemical nature of the direct ligands and the structure of the surrounding hydrogen bond network are crucial for both the activity of carbonic anhydrase and the metal ion affinity of the zinc-binding site. An understanding of naturally occurring zinc-binding sites will aid in creating de novo zinc-binding proteins and in designing new metal sites in existing proteins for novel purposes such as to serve as metal ion biosensors.
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Determination of Picomolar Concentrations of Metal Ions Using Fluorescence Anisotropy: Biosensing with a “Reagentless” Enzyme Transducer
Analytical Chemistry
Because of their high affinity and selectivity, metalloproteins can be used as transducers in novel sensors, i.e., biosensors, for the determination of trace levels of metal ions in solution. Here, we exploit carbonic anhydrase to determine picomolar to nanomolar concentrations of free transition metal ions by fluorescence anisotropy (polarization) in a reagentless format. Carbonic anhydrase variants engineered with a cysteine replacing a residue chosen near the active site (F131C and H64C)…
Because of their high affinity and selectivity, metalloproteins can be used as transducers in novel sensors, i.e., biosensors, for the determination of trace levels of metal ions in solution. Here, we exploit carbonic anhydrase to determine picomolar to nanomolar concentrations of free transition metal ions by fluorescence anisotropy (polarization) in a reagentless format. Carbonic anhydrase variants engineered with a cysteine replacing a residue chosen near the active site (F131C and H64C) were covalently labeled with derivatives of benzoxadiazole sulfonamide. These labeled variants exhibited changes in anisotropy up to 0.07 upon binding free Cu(II), Co(II), and Zn(II) with apparent Kd's close to the values observed with wild-type apocarbonic anhydrase. The covalent attachment of the label has significant advantages over noncovalent labels we have described previously. Furthermore, the metal ion-dependent anisotropy changes were predictable using simple theory. The results demonstrate that free transition metal ions can be determined at trace levels in aqueous solution using inexpensive instruments.
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Determination of multiple analytes using a fiber optic biosensor based on fluorescence energy transfer
SPIE Proceedings
Recently, we have developed a biosensor for zinc based on the very tight binding of this metal by the enzyme carbonic anhydrase, which requires Zn(II) for catalysis. We were able to transduce the binding of the metal as a change in fluorescence intensity or lifetime by use of a colored inhibitor whose metal-dependent binding permits fluorescence resonance energy transfer (Forster transfer) to occur. We have extended this concept to include other metals and other analytes which may be bound in…
Recently, we have developed a biosensor for zinc based on the very tight binding of this metal by the enzyme carbonic anhydrase, which requires Zn(II) for catalysis. We were able to transduce the binding of the metal as a change in fluorescence intensity or lifetime by use of a colored inhibitor whose metal-dependent binding permits fluorescence resonance energy transfer (Forster transfer) to occur. We have extended this concept to include other metals and other analytes which may be bound in the native (or mutant) enzyme active site with a concomitant color change; the color change is transduced as a change in energy transfer efficiency. We have also recently demonstrated a similar approach, wherein the presence of a metal ion in the binding site is transduced as a change in fluorescence anisotropy. Results in cuvettes and with fiber optic sensors are shown.
Patenter
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PROCESSES FOR PRODUCING FERMENTATION PRODUCTS
Udstedt EU WO/2006/101832
The present invention provides a process of producing a fermentation product comprises the steps of i) pre-treating lignocellulosic material to release or separate cellulose, hemi-cellulose and/or lignin, ii) subjecting the pre-treated material to a cellulase, iii) fermenting in the presence of a fermenting organism, wherein xylose isomerase is added in step ii) and/or step iii).
Andre opfindereSe patent -
COMPOSITIONS AND METHODS FOR ENHANCING PROTEIN EXPRESSION, SOLUBILITY, AND PURIFICATION
Anmeldt WSGR Docket Number 57607-711.101
Sprog
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English
Modersmåls- eller tosprogsfærdighed
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Danish
Professionel praktisk færdighed
Organisationer
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CphLAN
Member
– nuCphLAN is a Copenhagen-based Laboratory Automation Networking organization focused on helping fellow laboratory automation technicians, scientists, and managers succeed in their work by sharing experiences, tips, and honest opinions about various laboratory automation solutions.
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3 personer har anbefalet Keith
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From concept to stores, EQUII's Multi-Grain + Fiber Bread is trailblazing the path to sustainable protein and balanced nutrition on our plates…
From concept to stores, EQUII's Multi-Grain + Fiber Bread is trailblazing the path to sustainable protein and balanced nutrition on our plates…
Keith McCall synes godt om dette
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bright #Future ahead with more #biosolutions by #ChrHansen & #Novozymes as #novonesis https://lnkd.in/dYjxm6Wg
bright #Future ahead with more #biosolutions by #ChrHansen & #Novozymes as #novonesis https://lnkd.in/dYjxm6Wg
Keith McCall synes godt om dette
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Når dette år slutter er det et Farvel og på gensyn til AGC og alle mine fantastiske kolleger i AD. kæmpe tak til alle for de fantastiske år og gode…
Når dette år slutter er det et Farvel og på gensyn til AGC og alle mine fantastiske kolleger i AD. kæmpe tak til alle for de fantastiske år og gode…
Keith McCall synes godt om dette
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Thrilled to announce my new role as a Laboratory Technician in the QC Microbiology Department at AGC Biologics! Excited to be part of a fantastic…
Thrilled to announce my new role as a Laboratory Technician in the QC Microbiology Department at AGC Biologics! Excited to be part of a fantastic…
Keith McCall synes godt om dette
Andre lignende profiler
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Anne Ingeborg Myhr
NORCE Norwegian Research Centre AS
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Ani Tejirian, Biochemistry BS
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Elisabeth Ueland
Quality manager raw materials and suppliers, Skretting Norge
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Naouel Gharbi
Research Director Fish Biology & Aquaculture (FBA) Group
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Jeffrey Hern
Power Plant Operator at Northern California power agency
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Jannie Rosendahl Christensen
Laboratory Technician hos Samplix ApS
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Preparative Biochemistry and Biotechnology (PBB)
Taylor & Francis Journal
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Amber Clark
Human Resources Manager at Atlas Home Improvement
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Kate Boccadoro
Chief Scientist NORCE Norwegian Research Centre
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Paul Addis
Partner at Michigan Justice
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Andre med navnet Keith McCall
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Keith McCall
Assistant Director, Office of Graduate Fellowships and Awards
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Keith McCall
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Keith McCall
Assistant Director of Creative Video Northwestern Football
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Keith McCall
President and CEO at Dream City.Org (Dream City Charter School)
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