TY - JOUR A1 - Kunte, Hans-Jörg A1 - Lentzen, G. A1 - Galinski, E.A. T1 - Industrial production of the cell protectant ectoine: protection mechanisms, processes, and products N2 - Bacteria, Ärchaea and Eukatya can adapt to saline environments by accumulating compatible solutes in order to maintain an osmotic equilibrium. Compatible solutes are of diverse Chemical structure (sugars, polyols, amino acid derivatives) and are beneficial for bacterial cells not only as osmoregulatory solutes, but also as protectants of proteins by mitigating detrimental effects of ffeezing, drying and high temperatures. The aspartate derivative ectoine is a wide spread compatible solute in Bacteria and possesses additional protective properties compared with other compatible solutes, and stabilizes even vvhole cells against stresses such as UV radiation or cytotoxins. The protective properties of ectoine for proteins can be explained by its strong (kosmotropic) interaction with water and subsequent exclusion ffom Protein surface, the decrease of the solubility of the peptide backbone and the strengthening of intramolecular hydrogen bonds (secondary structures). The stabilizing and UV-protective properties of ectoine attracted industry, which saw the potential to market ectoine as a novel active component in health care products and cosmetics. In joint efforts of industry and research large-scale fermentation procedures have been developed with the halophilic bacterium Halomonas elongata used as a producer strain. The two key technologies that allow for the annual production of ectoine on a scale of tons are the bacterial milking procedure and the development and application of ectoine-excreting mutants (“leaky” mutant). The details of these two procedures including the strain development and fermentation processes will be introduced and current and future applications of ectoine will be discussed. KW - Bacterial milking KW - Batch fermentation KW - Continuous culture KW - Ectoine excretion KW - Hofmeister effect KW - Kosmotrope KW - Leaky mutant KW - Osmophobic effect KW - Protein protection KW - Preferential exclusion PY - 2014 SN - 2211-5501 SN - 2211-551X VL - 3 IS - 1 SP - 1 EP - 16 PB - Bentham Science CY - Sharjah [u.a.] AN - OPUS4-30078 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Brodehl, Antje A1 - Möller, Anne A1 - Kunte, Hans-Jörg A1 - Koch, Matthias A1 - Maul, Ronald T1 - Biotransformation of the mycotoxin zearalenone by fungi of the genera Rhizopus and Aspergillus N2 - Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin biosynthesized by various Fusarium fungi. These fungal species frequently infest grains; therefore, ZEN represents a common contaminant in cereal products. The biotransformation of ZEN differs significantly from species to species, and several metabolites are known to be formed by animals, plants, and microorganisms. The aim of the present study was to investigate the microbial conversion of ZEN by species of the genera Rhizopus and Aspergillus representing relevant fungi for food processing (e.g. fermentation). To monitor the ZEN metabolism, ZEN was added to liquid cultures of the different fungal species. After a period of 3 days, the media were analyzed by HPLC-MS/MS for metabolite formation. Two Aspergillus oryzae strains and all seven Rhizopus species were able to convert ZEN into various metabolites, including ZEN-14-sulfate as well as ZEN-O-14- and ZEN-O-16-glucoside. Microbial transformation of ZEN into the significantly more estrogenic α-zearalenol (α-ZEL) was also observed. Additionally, a novel fungal metabolite, α-ZEL-sulfate, was detected. Semi-quantification of the main metabolites indicates that more than 50% of initial ZEN may be modified. The results show that fungal strains have the potential to convert ZEN into various metabolites leading to a masking of the toxin, for example in fermented food. KW - Microbial conversion KW - Metabolites KW - Fermentation KW - Alpha-zearalenol KW - Conjugation KW - Mycotoxin biotransformation KW - Zearalenone-sulfate PY - 2014 U6 - https://doi.org/10.1111/1574-6968.12586 SN - 0378-1097 SN - 1574-6968 VL - 359 IS - 1 SP - 124 EP - 130 PB - Wiley-Blackwell CY - Malden, Mass., USA AN - OPUS4-31636 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Kunte, Hans-Jörg A1 - Lentzen, G. A1 - Galinski, E. T1 - Industrial production of the cell protectant ectoine: protection mechanisms, processes, and products N2 - Bacteria, Archaea and Eukarya can adapt to Saline environments by accumulating compatible solutes in order to maintain an osmotic equilibrium. Compatible solutes are of diverse chemical structure (sugars, polyols, amino acid derivatives) and are beneficial for bacterial cells not only as osmoregulatory solutes, but also as protectants of proteins by mitigating detrimental effects of freezing, drying and high temperatures. The aspartate derivative ectoine is a wide spread compatible solute in Bacteria and possesses additional protective properties compared with other compatible solutes, and stabilizes even whole cells against stresses such as UV radiation or cytotoxins. The protective properties of ectoine for proteins can be explained by its strong (kosmotropic) interaction with water and subsequent exclusion from Protein surface, the decrease of the solubility of the peptide backbone and the strengthening of intramolecular hydrogen bonds (secondary structures). The stabilizing and UV-protective properties of ectoine attracted industry, which saw the potential to market ectoine as a novel active component in health care products and cosmetics. In joint efforts of industry and research large-scale fermentation procedures have been developed with the halophilic bacterium Halomonas elongata used as a producer strain. The two key technologies that allow for the annual production of ectoine on a scale of tons are the bacterial milking procedure and the development and application of ectoine-excreting mutants (“leaky” mutant). The details of these two procedures including the strain development and Fermentation processes will be introduced and current and future applications of ectoine will be discussed. KW - Bacterial milking KW - Batch fermentation KW - Continuous culture KW - Ectoine excretion KW - Hofmeister effect KW - Kosmotrope KW - “leaky” mutant KW - Osmophobic effect KW - Protein protection KW - Preferential exclusion PY - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-376805 SN - 2211-551X VL - 3 IS - 1 SP - 10 EP - 25 PB - Bentham Science AN - OPUS4-37680 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Kuhne, Maren A1 - Dippong, Martin A1 - Flemig, Sabine A1 - Hoffmann, Katrin A1 - Petsch, K. A1 - Schenk, J.A. A1 - Kunte, Hans-Jörg A1 - Schneider, Rudolf T1 - Comparative characterization of mAb producing hapten-specific hybridoma cells by flow cytometric analysis and ELISA N2 - A novel method that optimizes the screening for antibody-secreting hapten-specific hybridoma cells by using flow cytometry is described. Cell clones specific for five different haptens were analyzed. We selectively double stained and analyzed fixed hybridoma cells with fluorophore-labeled haptens to demonstrate the target-selectivity, and with a fluorophore-labeled anti-mouse IgG antibody to characterize the level of surface expression of membrane-bound IgGs. ELISA measurements with the supernatants of the individual hybridoma clones revealed that antibodies from those cells, which showed the highest fluorescence intensities in the flow cytometric analysis, also displayed the highest affinities for the target antigens. The fluorescence intensity of antibody-producing cells corresponded well with the produced antibodies' affinities toward their respective antigens. Immunohistochemical staining verified the successful double labeling of the cells. Our method makes it possible to perform a high-throughput screening for hybridoma cells, which have both an adequate IgG production rate and a high target affinity. KW - Immunization KW - Hapten KW - Monoclonal antibodies KW - Hybridoma KW - Flow cytometry KW - ELISA KW - Estradiol KW - Estrone KW - Digoxigenin KW - Zearalenone KW - Aflatoxin KW - CLSM PY - 2014 U6 - https://doi.org/10.1016/j.jim.2014.07.004 SN - 0022-1759 SN - 1872-7905 VL - 413 SP - 45 EP - 56 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-32322 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -