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WORKSHOP „Nutrition-related biotechnology“. Improving food production by biotechnology. Prof. Wilfried Schwab Biotechnology of Natural Products Technical University München Liesel -Beckmann-Str. 1 Germany. Hamburg. Berlin. Germany. Frankfurt. Freising. München.
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WORKSHOP „Nutrition-related biotechnology“ Improving food production by biotechnology Prof. Wilfried Schwab Biotechnology of Natural Products Technical University München Liesel-Beckmann-Str. 1 Germany Hamburg Berlin Germany Frankfurt Freising München Prof. Dr. Wilfried Schwab
TUM The Entrepreneurial University TUM was one of the first "Universities of Excellence" of the nationwide Excellence Initiative announced by DFG TUM educates approximately 25,000 students in 133 study programs. Its scientists teach and research across multiple disciplines. Mathematics Physics Chemistry TUM School of Management Civil Engineering and Surveying Architecture Mechanical Engineering Electrical Engineering and Information Technology Informatics Center of Life and Food Sciences Weihenstephan Medicine Sport and Health Science TUM School of Education airport Prof. Dr. Wilfried Schwab
2003 Prof. Dr. Wilfried Schwab
Campus Weihenstephan in Freising Research Departments at Center of Life and Food Sciences Weihenstephan Agricultural Economics Animal Sciences Biosciences Ecology and Ecosystem Management Engineering Sciences for Food Products and Biogenetic Raw Materials (Life Science Engineering) Nutrition and Food Sciences Plant Sciences dedicated to “Nutrition-related biotechnology” Prof. Dr. Wilfried Schwab
BIOTECHNOLOGY OF NATURAL PRODUCTS founded in 2003 asDegussaprofessorship Biomolecular Food Technology Mission: to study the biosynthetic pathways and the metabolism of plant secondary products during fruit development by analytical, biochemical and molecular biological techniques Goal: To develop biocatalytical processes for the production of natural compounds which determine food quality due to their technological or physiological effects Prof. Dr. Wilfried Schwab
On-goingcollaborationwithChileanresearchers Raúl Herrera Faúndez Bioquímico, Ph.D. InstitutoBiologíaVegetal y BiotecnologíaUniversidad de Talca 2 Norte 685 phone 56-71-200280 fax 56-71-200276 Miriam E González Rojas; PhDstudent Instituto de BiologíaVegetal y Biotecnología Universidad de Talca Casilla 747, Talca, Chile Fono: +56-71-200275 Fax: +56-71-200276 DAAD fundedresearcherexchange 15. September 2010 – 20. May 2011 Project: Biochemical characterization of FcAAT1 an enzyme involved in the formation of fruit esters in strawberry FcAAT1 Prof. Dr. Wilfried Schwab
Improving food production by biotechnology 1 production of high-value food crops (biotechnology as analytical tool or as method) quantitative: higher biomass through resistance to diseases and pests and environmental stresses (drought, salt); higher efficiency of photosynthesis qualitative: vitamins, essential minerals, protein content, allergens and anti-nutrients, altered starch and fatty acid profile, antioxidant (lycopene, polyphenols) 2 biotechnology to improve microorganisms that are used for the production of food (bread, beer, wine, starter culture for meat products, probiotics) 3 bioprocess for the production of food additives (vitamins, amino acids, flavor, sweetners, carotenoids, modified starch, etc) 4 biotechnological production of enzymes used for food industry (alpha-amylase, glucose isomerase, chymosin, pectinase, lipase, lipoxygenase, etc.) Prof. Dr. Wilfried Schwab
Improving food production by biotechnology 1 production of high-value food crops (biotechnology as analytical tool or as method) quantitative: higher biomass through resistance to diseases and pests and environmental stresses (drought, salt); higher efficiency of photosynthesis qualitative: vitamins, essential minerals, protein content, allergens and anti-nutrients, altered starch and fatty acid profile, antioxidant (lycopene, polyphenols) 2 biotechnology to improve microorganisms that are used for the production of food (bread, beer, wine, starter culture for meat products, probiotics) 3 bioprocess for the production of food additives (vitamins, amino acids, flavor, sweetners, carotenoids, modified starch, etc) 4 biotechnological production of enzymes used for food industry (alpha-amylase, glucose isomerase, chymosin, pectinase, lipase, lipoxygenase, etc.) Prof. Dr. Wilfried Schwab
Strawberry Bet v 1-homologous allergen (Fra a 1) is down-regulated in colorless strawberry mutant Hjernø et al., Proteomics 2006, 6, 1574-1587 Prof. Dr. Wilfried Schwab
A B C Intron F D E 14 days Functionalgenomics in fruit: Transient overexpressionorRNAi-mediatedgenesilencing in ripeningstrawberryfruit chalcone synthase (CHS) Agroinfiltration Hoffmann et al. Plant J. 2006, 48, 818 Prof. Dr. Wilfried Schwab
Fra a 1 Homologue of Bet v1 in strawberry Nucleotides alignment Proteins alignment m109b07R Fra a 1e c201c06R Fra a 2 m117g04R Fra a 3 Munoz et al., Molecular Plant 2010, 3, 113 Prof. Dr. Wilfried Schwab
RT-PCR analysis of Fra a 1, Fra a 2 and Fra a 3 in strawberry Fra a 1 RT-PCR reactions. • Were performed with primers specific for the ESTs M109B07, C201C06 or M117G04. • The interspacer gene was used as control. Fra a 2 • RNA samples were from the F. ananassa vegetative tissues leaves (L), roots (Rt) and closed (CF) or fully mature flowers (OF), as well as from green (G), white (W), red (R) or post-harvest (PH) fruits Fra a 3 Prof. Dr. Wilfried Schwab
Positive control (CHSi) Fra a 1 Negative control (pBI-intron) Fra a 3 Downregulationofthestrawberry allergen Fra a Prof. Dr. Wilfried Schwab
Effects of Fra a Fra a Phe PAL allergen CHS coumaroyl CoA flavonoids coniferyl alcohol pathogenesis- related (PR) protein anthocyanins lignin phenylpropenoids benzenoids resistance color firmness flavor agronomic traits Prof. Dr. Wilfried Schwab
Improving food production by biotechnology 1 production of high-value food crops (biotechnology as analytical tool or as method) quantitative: higher biomass through resistance to diseases and pests and environmental stresses (drought, salt); higher efficiency of photosynthesis qualitative: vitamins, essential minerals, protein content, allergens and anti-nutrients, altered starch and fatty acid profile, antioxidant (lycopene, polyphenols) 2 biotechnology to improve microorganisms that are used for the production of food (bread, beer, wine, starter culture for meat products, probiotics) 3 bioprocess for the production of food additives (vitamins, amino acids, flavor, sweetners, carotenoids, modified starch, etc) 4 biotechnological production of enzymes used for food industry (alpha-amylase, glucose isomerase, chymosin, pectinase, lipase, lipoxygenase, etc.) Prof. Dr. Wilfried Schwab
Metabolism of proteins, peptides and amino acids Meat proteins Lactobacillus sakei genome: lack of aminotransferase and decarboxylase genes new protein with aminotransferase activity isolated peptides rapidly disappear from the solution but amino acids produce higher levels of aroma compounds after resorption peptides are not only hydrolyzed to amino acids ? Prof. Dr. Wilfried Schwab
Improving food production by biotechnology 1 production of high-value food crops (biotechnology as analytical tool or as method) quantitative: higher biomass through resistance to diseases and pests and environmental stresses (drought, salt); higher efficiency of photosynthesis qualitative: vitamins, essential minerals, protein content, allergens and anti-nutrients, altered starch and fatty acid profile, antioxidant (lycopene, polyphenols) 2 biotechnology to improve microorganisms that are used for the production of food (bread, beer, wine, starter culture for meat products, probiotics) 3 bioprocess for the production of food additives (vitamins, amino acids, flavor, sweetners, carotenoids, modified starch, etc) 4 biotechnological production of enzymes used for food industry (alpha-amylase, glucose isomerase, chymosin, pectinase, lipase, lipoxygenase, etc.) Prof. Dr. Wilfried Schwab
Biosynthesis of thekeystrawberryflavorcompound (HDMF) Raab et al., Plant Cell, 2006, 18, 1023 Klein et al., J. Agric. Food Chem., 2007, 55, 6705 Wein et al., Plant J., 2002, 31, 755 Lavid et al., J. Agric. Food Chem., 2002, 50, 4025 Lunkenbein et al., J. Exp. Bot., 2006, 57, 4025 Prof. Dr. Wilfried Schwab
Biotechnological production of norisoprenoids Prof. Dr. Wilfried Schwab
Improving food production by biotechnology 1 production of high-value food crops (biotechnology as analytical tool or as method) quantitative: higher biomass through resistance to diseases and pests and environmental stresses (drought, salt); higher efficiency of photosynthesis qualitative: vitamins, essential minerals, protein content, allergens and anti-nutrients, altered starch and fatty acid profile, antioxidant (lycopene, polyphenols) 2 biotechnology to improve microorganisms that are used for the production of food (bread, beer, wine, starter culture for meat products, probiotics) 3 bioprocess for the production of food additives (vitamins, amino acids, flavor, sweetners, carotenoids, modified starch, etc) 4 biotechnological production of enzymes used for food industry (alpha-amylase, glucose isomerase, chymosin, pectinase, lipase, lipoxygenase,dioxygenases) Prof. Dr. Wilfried Schwab
‘Green note‘ production 13-LOX 9-LOX 9-HPL 13-HPL C9 Volatile Products Nonenals, Nonadienals Hexanal, Hexenals Prof. Dr. Wilfried Schwab
Transient expression of LOX and HPL genes Prof. Dr. Wilfried Schwab
Proteins relatedtofoodindustry Flavor FaOMT, FaQR furaneol (strawberry) FaAAT1, FcAAT fruitester FaCCD1, RdCCD1 norisoprenoid, eg. b-ionone FaNES, FaPIN, LaLIN, LaBER, LaLIM mono- andsesquiterpenes FaEGS phenylpropenes, eg. eugenol FaJMT methyljasmonate CrSMT S-methyltransferase thiomethylether TbLOX fattyacidhydroperoxide aldehydes Pigment FaGT1 anthocyan FaGT6, 7 flavonoid FaCHS, FaANS, FaDFR, FaANR anthocyan Firmness shelflife FaCCR, FaCAD, FaPOD lignin Pathogene resistance allergenicity Fra a 1,2,3 affectsanthocyanformation, allergens FaGT2, 5 hydroxycinnamoylglucose Prof. Dr. Wilfried Schwab
Phytoenzymes Towards functionalized natural products through exploitation of the plant gene pool plants are underrepresented as source for industrial biocatalysts: dioxygenases Plants produce an amazing diversity of low molecular weight compounds making the plant proteome a rich source of remarkable biocatalysts such as dioxygenases that catalyze hydroxylation, epoxidation, and peroxidation reactions in a regio- and enantioselective manner. The goal of this interdisciplinary Research Unit is the detailed study of a set of plant dioxygenases of hitherto unknown function with respect to substrate/product specificity, catalytic activity, and protein structure. The project will provide novel tools for protein functional analysis, substantially improve gene annotation, expand the reactome and chemical space of dioxygenases and lead to a better understanding of their biological roles. The newly discovered enzymatic activities will likely also lead to new applications in bioorganic chemistry. Prof. Dr. Wilfried Schwab
THANKS TO and DFG, DAAD, Firmenich, Degussa, BMBF, EU, AIF, ZuTech for financial support Prof. Dr. Wilfried Schwab