A5016123662- Bacteriophages and microbial interactions
- Bacterial Genetics and Biotechnology
- Viral Infectious Diseases and Gene Expression in Insects
- Protein purification and stability
- Microbial Metabolic Engineering and Bioproduction
- RNA and protein synthesis mechanisms
- SARS-CoV-2 and COVID-19 Research
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- RNA Interference and Gene Delivery
- Cancer Research and Treatments
- Monoclonal and Polyclonal Antibodies Research
- Biofuel production and bioconversion
- Genomics and Phylogenetic Studies
- SARS-CoV-2 detection and testing
- Escherichia coli research studies
- Transgenic Plants and Applications
- Glycosylation and Glycoproteins Research
- Biosensors and Analytical Detection
- Endoplasmic Reticulum Stress and Disease
- Acute Kidney Injury Research
- Viral gastroenteritis research and epidemiology
- Microfluidic and Bio-sensing Technologies
- Biosimilars and Bioanalytical Methods
- Microfluidic and Capillary Electrophoresis Applications
Pharis Biotec (Germany)
2020-2023
Hudson Institute
2019
John Wiley & Sons (United States)
2019
BOKU University
2006-2016
Austrian Centre of Industrial Biotechnology (Austria)
2013-2014
Graz University of Technology
2012
Escherichia coli K–12 and B strains are among the most frequently used bacterial hosts for production of recombinant proteins on an industrial scale. To improve existing processes to accelerate bioprocess development, we performed a detailed host analysis. We investigated different behaviors E. BL21, RV308, HMS174 in response high-glucose concentrations. Tightly controlled cultivations were conducted under defined environmental conditions in-depth analysis physiological behavior. In addition...
The phage-derived T7 RNA polymerase is the most prominent orthogonal transcriptions system used in field of synthetic biology. However, gene expression driven by prone to read-through transcription due contextuality terminator. native terminator has a termination efficiency approximately 80% and therefore provides insufficient insulation unit. By using combination signal with two well-known transcriptional terminators (rrnBT1 T7), we have been able increase 99%. To characterize putative...
ABSTRACT Plasmid-based Escherichia coli BL21(DE3) expression systems are extensively used for the production of recombinant proteins. However, combination a high gene dosage with strong promoters exerts extremely stressful conditions on producing cells, resulting in multitude protective reactions and malfunctions host cell impact yield quality product. Here, we provide in-depth characterization plasmid-based perturbations protein production. A plasmid-free T7 system single copy interest...
Modulating resource allocation in bacteria to redirect metabolic building blocks the formation of recombinant proteins rather than biomass remains a grand challenge biotechnology. Here, we present novel approach for improved protein production (RPP) using Escherichia coli (E. coli) by decoupling synthesis from cell growth. We show that division and host mRNA transcription can be successfully inhibited coexpression bacteriophage-derived E. RNA polymerase (RNAP) inhibitor peptide genes...
Antibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection or vaccination. While first-generation antibody have primarily provided qualitative results, accurate seroprevalence studies and tracking of levels over time require highly specific, sensitive quantitative test setups.
Genetic modification of stem cells, prior to transplantation, can enhance their survival and improve function in cell therapy settings. Mesenchymal cells (MSC) are considered one the most promising tools for cell-based gene therapy, due multipotency, ease isolation, as well high ex vivo expansion potential. Neural (NSC) may also present an ideal route have been use replacement therapies various neurodegenerative diseases. Gene therapy-based applications require transfer genetic material,...
The presence of antibiotic resistance genes in the delivered plasmids is one drawbacks modern gene therapy and DNA vaccine applications. Here, we describe a strategy that allows for plasmid selection bacterial hosts, without requirement any marker. Several strains were modified, so plasmid's replicational inhibitor RNA I could suppress translation growth essential by RNA-RNA antisense reaction. An (murA) was modified such repressor protein (tetR) would hamper its expression. Only and, hence,...
Abstract Background Precise regulation of gene expression is utmost importance for the production complex membrane proteins (MP), enzymes or other toxic to host cell. In this article we show that genes under control a normally Isopropyl β- d -1-thiogalactopyranoside (IPTG)-inducible P T7-lacO promoter can be induced solely with l -arabinose in newly constructed Escherichia coli BL21-AI< gp2 >, strain based on recently published approach bacteriophage inspired growth-decoupled...
Plasmid DNA (pDNA) is a key biotechnological product whose importance became apparent in the last years due to its role as raw material messenger ribonucleic acid (mRNA) vaccine manufacturing process. In pharmaceutical production processes, cells need grow defined medium order guarantee highest standards of quality and repeatability. However, often these requirements result low titer, productivity, yield. this study, we used constraint-based metabolic modeling optimize average volumetric...
Abstract Sugar nucleotide‐dependent (Leloir) glycosyltransferases from plants are important catalysts for the glycosylation of small molecules and natural products. Limitations on their applicability biocatalytic synthesis arise because low protein expression (≤10 mg/L culture) in standard microbial hosts. Here, we showed two representative glycosyltransferases: sucrose synthase soybean UGT71A15 apple. A synthetic biology‐based strategy decoupling enzyme Escherichia coli BL21(DE3) cell...
The site-specific incorporation of noncanonical amino acids (ncAAs) into proteins by amber stop codon suppression has become a routine method in academic laboratories. This approach requires an suppressor tRNACUA to read the and aminoacyl-tRNA synthetase charge with ncAA. However, major drawback is low yield mutant protein comparison wild type. effect primarily results from competition release factor 1 charged for at A-site ribosome. A number laboratories have attempted improve efficiency...
In recent years, receptor-mediated drug delivery has gained major attention in the treatment of cancer. The pathogen-derived Shiga Toxin B subunit (STxB) can be used as a carrier that detects tumor-associated glycosphingolipid globotriaosylceramide (Gb3) receptors. While conjugation via lysine or cysteine offers random attachment to carriers, click chemistry potential improve engineering systems site specificity eliminate interference with active binding tumor ligands. We present production...
The secretion and direct capture of proteins from the extracellular medium is a promising approach for purification, thus enabling integrated bioprocesses.We demonstrate nanobody (VHH) to (EM) its by bare, non-functionalized magnetic nanoparticles (MNPs). An ompA signal peptide periplasmic localization, polyglutamate-tag (E8 ) selective MNP binding, factor Xa protease cleavage site were fused N-terminally nanobody. production E8 -VHH (36 mg L-1 was enabled using growth-decoupled Escherichia...
The use of plasmid DNA for gene therapeutical purposes is a novel technology with advantages and drawbacks. One the required improvements to avoid antibiotic resistance genes or other additional sequences selection within plasmid. Here, we describe an alternative approach equip ColE1 regulatory function cell, which could be used carrying cells. No are required, since mechanism based on RNA/RNA antisense interaction involving naturally occurring RNA I derived from plasmid's origin...
The nucleocapsid protein (NP) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for several steps the viral life cycle, and abundantly expressed during infection, making it an ideal diagnostic target protein. This has a strong tendency dimerization interaction with nucleic acids. For first time, high titers NP were in E. coli CASPON tag, using growth-decoupled expression system. Purification was accomplished by nuclease treatment cell homogenate sequence downstream...
Abstract With the growing interest in continuous cultivation of Escherichia coli , secretion product to medium is not only a benefit, but necessity future bioprocessing. In this study, it shown that induced decoupling growth and heterologous gene expression E. X‐press strain (derived from BL21(DE3)) facilitates extracellular recombinant protein production. The effect process parameters temperature specific glucose consumption rate ( q S ) on growth, productivity, lysis leakiness,...
Abstract Antibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection. While first-generation antibody have primarily provided qualitative results with low specificity, accurate seroprevalence studies and tracking of levels over time require highly specific, sensitive quantitative test setups. Here, we describe two ELISA based on the spike receptor-binding domain nucleocapsid protein. Comparative expression in bacterial, insect, mammalian plant-based...
Escherichia coli strain K-12 substrain HMS174 is an engineered descendant of the E. wild-type strain. Like its ancestor, it important organism in biotechnological research and used fermentation processes for heterologous protein production. Here, we report complete genome sequence (ATCC 47011).