Recombinant Production of Mussel Byssus Inspired Proteins

2018 ◽  
Vol 13 (12) ◽  
pp. 1800146 ◽  
Author(s):  
Jia Wang ◽  
Thomas Scheibel
Keyword(s):  
Recombinant Production ◽  
Mussel Byssus

The Medicinal Chemistry of Therapeutic Peptides: Recent Developments in Synthesis and Design Optimizations

2019 ◽  
Vol 26 (13) ◽  
pp. 2330-2355 ◽  
Author(s):  
Anutthaman Parthasarathy ◽  
Sasikala K. Anandamma ◽  
Karunakaran A. Kalesh
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Therapeutic Potential ◽  
The Past ◽  
Recombinant Production ◽  
Tremendous Progress ◽  
Recent Developments ◽  
Therapeutic Peptides ◽  
Development Processes ◽  
Delivery Strategies

Peptide therapeutics has made tremendous progress in the past decade. Many of the inherent weaknesses of peptides which hampered their development as therapeutics are now more or less effectively tackled with recent scientific and technological advancements in integrated drug discovery settings. These include recent developments in synthetic organic chemistry, high-throughput recombinant production strategies, highresolution analytical methods, high-throughput screening options, ingenious drug delivery strategies and novel formulation preparations. Here, we will briefly describe the key methodologies and strategies used in the therapeutic peptide development processes with selected examples of the most recent developments in the field. The aim of this review is to highlight the viable options a medicinal chemist may consider in order to improve a specific pharmacological property of interest in a peptide lead entity and thereby rationally assess the therapeutic potential this class of molecules possesses while they are traditionally (and incorrectly) considered ‘undruggable’.

Enhanced Biocatalytic Activity of Recombinant Lipase Immobilized on Gold Nanoparticles

2019 ◽  
Vol 20 (6) ◽  
pp. 497-505 ◽  
Author(s):  
Abeer M. Abd El-Aziz ◽  
Mohamed A. Shaker ◽  
Mona I. Shaaban
Keyword(s):  
Gold Nanoparticles ◽  
Lipolytic Activity ◽  
Market Demand ◽  
Biotechnological Applications ◽  
Lipase Gene ◽  
Expression Plasmid ◽  
E Coli ◽  
Nickel Affinity Chromatography ◽  
Recombinant Production ◽  
Sds Page

Background: Bacterial lipases especially Pseudomonas lipases are extensively used for different biotechnological applications. Objectives: With the better understanding and progressive needs for improving its activity in accordance with the growing market demand, we aimed in this study to improve the recombinant production and biocatalytic activity of lipases via surface conjugation on gold nanoparticles. Methods: The full length coding sequences of lipase gene (lipA), lipase specific foldase gene (lipf) and dual cassette (lipAf) gene were amplified from the genomic DNA of Pseudomonas aeruginosa PA14 and cloned into the bacterial expression vector pRSET-B. Recombinant lipases were expressed in E. coli BL-21 (DE3) pLysS then purified using nickel affinity chromatography and the protein identity was confirmed using SDS-PAGE and Western blot analysis. The purified recombinant lipases were immobilized through surface conjugation with gold nanoparticles and enzymatic activity was colorimetrically quantified. Results: Here, two single expression plasmid systems pRSET-B-lipA and pRSET-B-lipf and one dual cassette expression plasmid system pRSET-B-lipAf were successfully constructed. The lipolytic activities of recombinant lipases LipA, Lipf and LipAf were 4870, 426 and 6740 IUmg-1, respectively. However, upon immobilization of these recombinant lipases on prepared gold nanoparticles (GNPs), the activities were 7417, 822 and 13035 IUmg-1, for LipA-GNPs, Lipf-GNPs and LipAf-GNPs, respectively. The activities after immobilization have been increased 1.52 and 1.93 -fold for LipA and LipAf, respectively. Conclusion: The lipolytic activity of recombinant lipases in the bioconjugate was significantly increased relative to the free recombinant enzyme where immobilization had made the enzyme attain its optimum performance.

scholarly journals Recombinant Production and Characterization of an Extracellular Subtilisin-Like Serine Protease from Acinetobacter baumannii of Fermented Food Origin

2021 ◽  
Author(s):  
Nur Syafiqah Muhammed ◽  
Nurulfarhana Hussin ◽  
Aik Siang Lim ◽  
Mohd Anuar Jonet ◽  
Shaza Eva Mohamad ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Serine Protease ◽  
Fermented Food ◽  
Recombinant Production

scholarly journals Protein L—More Than Just an Affinity Ligand

Processes ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 874
Author(s):  
Stefan Kittler ◽  
Mihail Besleaga ◽  
Julian Ebner ◽  
Oliver Spadiut
Keyword(s):  
Antibody Fragments ◽  
Downstream Process ◽  
Affinity Ligands ◽  
The Past ◽  
Affinity Ligand ◽  
Recombinant Production ◽  
Production Processes ◽  
Monitoring Tools ◽  
Platform Technology ◽  
Analytical Tools

In the past 30 years, highly specific drugs, known as antibodies, have conquered the biopharmaceutical market. In addition to monoclonal antibodies (mAbs), antibody fragments are successfully applied. However, recombinant production faces challenges. Process analytical tools for monitoring and controlling production processes are scarce and time-intensive. In the downstream process (DSP), affinity ligands are established as the primary and most important step, while the application of other methods is challenging. The use of these affinity ligands as monitoring tools would enable a platform technology to monitor process steps in the USP and DSP. In this review, we highlight the current applications of affinity ligands (proteins A, G, and L) and discuss further applications as process analytical tools.

scholarly journals Human Plasma and Recombinant Hemopexins: Heme Binding Revisited

2021 ◽  
Vol 22 (3) ◽  
pp. 1199
Author(s):  
Elena Karnaukhova ◽  
Catherine Owczarek ◽  
Peter Schmidt ◽  
Dominik J. Schaer ◽  
Paul W. Buehler
Keyword(s):  
Protein S ◽  
Soret Band ◽  
Cell System ◽  
Size Exclusion ◽  
Heme Binding ◽  
Systematic Analysis ◽  
Molar Ratios ◽  
Recombinant Production ◽  
Wide Range ◽  
Exclusion Chromatography

Plasma hemopexin (HPX) is the key antioxidant protein of the endogenous clearance pathway that limits the deleterious effects of heme released from hemoglobin and myoglobin (the term “heme” is used in this article to denote both the ferrous and ferric forms). During intra-vascular hemolysis, heme partitioning to protein and lipid increases as the plasma concentration of HPX declines. Therefore, the development of HPX as a replacement therapy during high heme stress could be a relevant intervention for hemolytic disorders. A logical approach to enhance HPX yield involves recombinant production strategies from human cell lines. The present study focuses on a biophysical assessment of heme binding to recombinant human HPX (rhHPX) produced in the Expi293FTM (HEK293) cell system. In this report, we examine rhHPX in comparison with plasma HPX using a systematic analysis of protein structural and functional characteristics related to heme binding. Analysis of rhHPX by UV/Vis absorption spectroscopy, circular dichroism (CD), size-exclusion chromatography (SEC)-HPLC, and catalase-like activity demonstrated a similarity to HPX fractionated from plasma. In particular, the titration of HPX apo-protein(s) with heme was performed for the first time using a wide range of heme concentrations to model HPX–heme interactions to approximate physiological conditions (from extremely low to more than two-fold heme molar excess over the protein). The CD titration data showed an induced bisignate CD Soret band pattern typical for plasma and rhHPX versions at low heme-to-protein molar ratios and demonstrated that further titration is dependent on the amount of protein-bound heme to the extent that the arising opposite CD couplet results in a complete inversion of the observed CD pattern. The data generated in this study suggest more than one binding site in both plasma and rhHPX. Furthermore, our study provides a useful analytical platform for the detailed characterization of HPX–heme interactions and potentially novel HPX fusion constructs.

scholarly journals Recombinant production of a peroxidase-protein G fusion protein in Pichia pastoris

2016 ◽  
Vol 219 ◽  
pp. 24-27 ◽  
Author(s):  
Florian Wolfgang Krainer ◽  
Barbara Darnhofer ◽  
Ruth Birner-Gruenberger ◽  
Anton Glieder
Keyword(s):  
Pichia Pastoris ◽  
Fusion Protein ◽  
Protein G ◽  
Recombinant Production

Recombinant Production of Isotope-Labeled Peptides and Spontaneous Cyclization of Amino-Terminal Glutamine into Pyroglutamic Acid

ChemBioChem ◽  
2012 ◽  
Vol 13 (10) ◽  
pp. 1421-1423 ◽  
Author(s):  
André Mischo ◽  
Oliver Ohlenschläger ◽  
Karl-Heinz Gührs ◽  
Matthias Görlach
Keyword(s):  
Pyroglutamic Acid ◽  
Amino Terminal ◽  
Recombinant Production

In SituAnalysis of Peptidyl DOPA in Mussel Byssus Using Rotational-Echo Double-Resonance NMR

1996 ◽  
Vol 333 (1) ◽  
pp. 221-224 ◽  
Author(s):  
Christopher A. Klug ◽  
Luis A. Burzio ◽  
J.Herbert Waite ◽  
Jacob Schaefer
Keyword(s):  
Double Resonance ◽  
Rotational Echo Double Resonance ◽  
Mussel Byssus

scholarly journals Recombinant Production of Human Aquaporin-1 to an Exceptional High Membrane Density in Saccharomyces cerevisiae

PLoS ONE ◽  
2013 ◽  
Vol 8 (2) ◽  
pp. e56431 ◽  
Author(s):  
Julie Bomholt ◽  
Claus Hélix-Nielsen ◽  
Peter Scharff-Poulsen ◽  
Per Amstrup Pedersen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Aquaporin 1 ◽  
Recombinant Production ◽  
Membrane Density
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