Affiliation: Vienna University of Technology
- Spadiut O, Rossetti L, Dietzsch C, Herwig C. Purification of a recombinant plant peroxidase produced in Pichia pastoris by a simple 2-step strategy. Protein Expr Purif. 2012;86:89-97 pubmed publisher..Combining a hydrophobic charge induction chromatography operated in flowthrough mode and a size-exclusion chromatography, we were able to purify rHRP more than 12-fold from a specific activity of 80U/mg to more than 1000U/mg. ..
- Krainer F, Capone S, JÃ¤ger M, Vogl T, Gerstmann M, Glieder A, et al. Optimizing cofactor availability for the production of recombinant heme peroxidase in Pichia pastoris. Microb Cell Fact. 2015;14:4 pubmed publisher..pastoris can be easily enhanced by supplementation of the cultivation medium with hemin. Thereby, secreted apo-species of the target protein are effectively saturated with cofactor, maximizing the yield of target enzyme activity. ..
- Slouka C, Wurm D, Brunauer G, Welzl Wachter A, Spadiut O, Fleig J, et al. A Novel Application for Low Frequency Electrochemical Impedance Spectroscopy as an Online Process Monitoring Tool for Viable Cell Concentrations. Sensors (Basel). 2016;16: pubmed..It is shown that impedance spectroscopy at low frequencies is a powerful tool for cultivation monitoring. ..
- Hausjell J, Halbwirth H, Spadiut O. Recombinant production of eukaryotic cytochrome P450s in microbial cell factories. Biosci Rep. 2018;38: pubmed publisher..This review describes current strategies for expression of P450s in the microbial cell factories Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris. ..
- Capone S, Ä†orajeviÄ‡ L, Bonifert G, Murth P, Maresch D, Altmann F, et al. Combining Protein and Strain Engineering for the Production of Glyco-Engineered Horseradish Peroxidase C1A in Pichia pastoris. Int J Mol Sci. 2015;16:23127-42 pubmed publisher..This variant shows considerable catalytic activity and thermal stability and is less glycosylated, which might allow more controlled and efficient conjugation to antibodies and lectins. ..
- Kopp J, Slouka C, Ulonska S, Kager J, Fricke J, Spadiut O, et al. Impact of Glycerol as Carbon Source onto Specific Sugar and Inducer Uptake Rates and Inclusion Body Productivity in E. coli BL21(DE3). Bioengineering (Basel). 2017;5: pubmed publisher..Additionally, it was shown that glycerol has beneficial effects on viability of cells and on productivity of the recombinant protein compared to glucose. ..
- Hofer A, Herwig C, Spadiut O. Lecithin is the key material attribute in soy bean oil affecting filamentous bioprocesses. AMB Express. 2018;8:90 pubmed publisher..Thus, over-dosage and potential resulting negative impacts on fungal growth and productivity can be omitted. ..
- Rajamanickam V, Metzger K, Schmid C, Spadiut O. A novel bi-directional promoter system allows tunable recombinant protein production in Pichia pastoris. Microb Cell Fact. 2017;16:152 pubmed publisher..This bi-directional promoter system offers a high degree of freedom for bioprocess design and development, making bi-directional promoters in P. pastoris highly attractive for recombinant protein production. ..
- Gundinger T, Spadiut O. A comparative approach to recombinantly produce the plant enzyme horseradish peroxidase in Escherichia coli. J Biotechnol. 2017;248:15-24 pubmed publisher..coli allowing straight forward screening for improved variants, and finally produce these variants as IB in high amounts, which are then refolded. ..
- Rajamanickam V, Wurm D, Slouka C, Herwig C, Spadiut O. A novel toolbox for E. coli lysis monitoring. Anal Bioanal Chem. 2017;409:667-671 pubmed publisher..coli lysis in a fast and reliable manner. We are convinced that this toolbox will not only facilitate E. coli bioprocess monitoring but will also allow enhanced process control in the future. ..
- Wurm D, Veiter L, Ulonska S, Eggenreich B, Herwig C, Spadiut O. The E. coli pET expression system revisited-mechanistic correlation between glucose and lactose uptake. Appl Microbiol Biotechnol. 2016;100:8721-9 pubmed publisher..We believe that this mechanistic model might serve as platform knowledge for E. coli. ..
- Eggenreich B, Scholz E, Wurm D, Forster F, Spadiut O. The production of a recombinant tandem single chain fragment variable capable of binding prolamins triggering celiac disease. BMC Biotechnol. 2018;18:30 pubmed publisher..The high binding specificity of tscFv towards prolamin-containing grains makes this novel molecule a valuable candidate to support patients suffering from CD in the future. ..
- Gmeiner C, Saadati A, Maresch D, Krasteva S, Frank M, Altmann F, et al. Development of a fed-batch process for a recombinant Pichia pastoris Î”och1 strain expressing a plant peroxidase. Microb Cell Fact. 2015;14:1 pubmed publisher..In terms of productivity and product purity a temperature of 20Â°C was highly beneficial. In summary, we determined cultivation conditions for a recombinant P. pastoris Î”och1 strain allowing high productivity and product purity. ..