Das Projekt "Teilprojekt A" wird vom Umweltbundesamt gefördert und von Leibniz Universität Hannover, Institut für Lebensmittelchemie durchgeführt. Aim: Valorisation of side-streams of the Citrus industry using the genetic diversity of monokarya from the basidiomycete Pleurotus sapidus. The genetic diversity of the basidiospores of Pleurotus sapidus (MKs) obtained from two dikaryotic strains of P. sapidus (Dk421 and Dk3174) will be exploited. Mks with high growth rate on milled Citrus peel, pulp and seed of orange, tangerine, lemon will be selected and grown as solid state and submerged fermentation (SF). Metabolites will be extracted and evaluated for biological activities. Samples before and after the fungal transformation taken from SSF and SF cultures will be analysed. Rapid product analyses using TLC and established coupled HPLC-DAD-ELSD will focus on the most promising strains. Specific targets are flavonoids with an increased number of hydroxyl groups on the B-ring, unsaturated carbonyls and terpenoids from the oxo-functionalisation of limonene, citronellal and farnesene isomers. High resolution and multi-dimensional GC-MS and multireaction monitoring (varying MS collision energies) will be used. Extracts from various strain/culture combinations (SSF or SF) will be lyophilized. One fraction of each sample will be tested for its biopesticide action, and another one for its quality as a feed supplement. SSF will be carried out in a rotary drum solid-substrate fermentation system.
The project is comprised of seven major work packages: 1. Generation and selection of the monokaryons (CITER) 2. Growth of the monokaryons (CITER) 3. Selection of the optimal culture conditions to obtain bioactive compounds using the selected Mk form step 2. (CITER, LUH, JLU, JUB) 4. Analytical evaluation of the biotransformation/conversion products (LUH, JLU) 5. Automated screening of Mks by chiral GC-GC (JLU) 6. Bioactivity test of crude extracts obtained from SSF and SF (IMBIV, IIB) 7. Bioprocess design and scale-up (JLU, JUB).
Das Projekt "Teilprojekt C" wird vom Umweltbundesamt gefördert und von Jacobs University Bremen GmbH, School of Engineering and Science durchgeführt. Valorisation of side-streams of the Citrus industry using the genetic diversity of monokarya from the basidiomycete Pleurotus sapidus. The genetic diversity of the basidiospores of Pleurotus sapidus (MKs) obtained from two dikaryotic strains of P. sapidus (Dk421 and Dk3174) will be exploited. Mks with high growth rate on milled Citrus peel, pulp and seed of orange, tangerine, lemon will be selected and grown as solid state and submerged fermentation (SF). Metabolites will be extracted and evaluated for biological activities. Samples before and after the fungal transformation taken from SSF and SF cultures will be analysed. Rapid product analyses using TLC and established coupled HPLC-DAD-ELSD will focus on the most promising strains. Specific targets are flavonoids with an increased number of hydroxyl groups on the B-ring, -- or -- unsaturated carbonyls, and terpenoids from the oxo-functionalisation of limonene, citronellal and farnesene isomers. High resolution and multi-dimensional GC-MS and multireaction monitoring (varying MS collision energies) will be used. Extracts from various strain/culture combinations (SSF or SF) will be lyophilized and milled. One fraction of each sample will be tested for its biopesticide action, and another one for its quality as feed supplement. Five and 150 L fermenters will be operated to scale-up the results. SSF will be carried out in a rotary drum solid-substrate fermentation system. The project is comprised of seven major work packages: 1. Generation and selection of the monokaryons (CITER) 2. Growth of the monokaryons (CITER) 3. Selection of the optimal culture conditions to obtain bioactive compounds using the selected Mk form step 2. (CITER, LUH, JLU, JUB) 4. Analytical evaluation of the biotransformation/conversion products (LUH, JLU) 5. Automated screening of Mks by chiral GC-GC (JLU) 6. Bioactivity test of crude extracts obtained from SSF and SF (IMBIV, IIB) 7. Bioprocess design and scale-up (JLU, JUB)
Das Projekt "Teilprojekt B" wird vom Umweltbundesamt gefördert und von Universität Gießen, Institut für Lebensmittelchemie und Lebensmittelbiotechnologie durchgeführt. Großvolumige Nebenströme der industriellen Citrus-Verarbeitung werden mit Basidiomyceten fermentiert, um neben Pilzbiomasse hochwertige Metabolite wie Aromen und funktionelle Lebensmittelinhaltsstoffe zu gewinnen. Monokaryotische Stämme (Mks) des Basidiomyceten Pleurotus sapidus werden auf Schrägagar direkt in Headspace-Vials auf dem Multi-Purpose-Sampler eines Gaschromatographiesystems kultiviert und vollautomatisiert mittels GC-Tandemmassenspektrometrie analysiert. Interessante Stämme werden anschließend zusätzlich olfaktometrisch mittels GC-MS/MS-O untersucht. Die potentesten Stämme werden in Emers- und Submerskulturen überführt und die jeweils gebildeten Aromastoffe qualitativ und quantitativ erfasst. Nichtflüchtige Aromapräkursoren werden zudem hochsensitiv mittels UHPLC-MS/MS detektiert. Mittels multidimensionaler Gaschromatographie in Kopplung mit einem massenspektrometrischen Detektor werden chirale Verbindungen getrennt und charakterisiert. Um den Partnern hinreichende Mengen an Probenmaterial für die Testung der insektiziden und fungiziden Aktivität ebenso wie für die Fütterungsexperimente mit Fischen zur Verfügung stellen zu können, ist eine weitere Maßstabsvergrößerung des Fermentationsprozesses ebenso erforderlich wie die Entwicklung eines geeigneten Downstream-Processings. Am LCB werden dafür Submerskulturen im 150 L-Bioreaktor mit 100 L Medium geführt. Kulturüberstand und Myzel werden mittels Filtration voneinander getrennt, das Myzel eingefroren, ggf. zur Standardisierung lyophilisiert und den Projektpartnern zur Verfügung gestellt.