technische universität münchen prof. dr. wilfried schwab improving food production by...

Technische Universität München

Prof. Dr. Wilfried Schwab

Improving food production by biotechnologyProf. Wilfried Schwab Biotechnology of Natural ProductsTechnical University MünchenLiesel-Beckmann-Str. 1Germany

WORKSHOP „Nutrition-related biotechnology“

Germany

Freising

München

Berlin

Frankfurt

Hamburg


airport

TUM was one of the first "Universities of Excellence" of the nationwide Excellence Initiative announced by DFGTUM educates approximately 25,000 students in 133 study programs. Its scientists teach and research across multiple disciplines.

MathematicsPhysics

ChemistryTUM School of Management

Civil Engineering and SurveyingArchitecture

Mechanical EngineeringElectrical Engineering and Information

TechnologyInformatics

Center of Life and Food Sciences Weihenstephan

MedicineSport and Health ScienceTUM School of Education

TUM The Entrepreneurial University




2003



Research Departments at Center of Life and Food Sciences WeihenstephanAgricultural Economics Animal Sciences BiosciencesEcology and Ecosystem ManagementEngineering Sciences for Food Products and Biogenetic Raw Materials (Life Science Engineering) Nutrition and Food Sciences Plant Sciences

dedicated to “Nutrition-related biotechnology”

Campus Weihenstephan in Freising



BIOTECHNOLOGY OF NATURAL PRODUCTS

Mission:to study the biosynthetic pathways

and the metabolism of plant secondary products during fruit

development by analytical, biochemical and molecular biological techniquesGoal:

To develop biocatalytical processes for the production of natural

compounds which determine food quality due to their technological or

physiological effects

founded in 2003 as Degussa professorship Biomolecular Food Technology



Raúl Herrera FaúndezBioquímico, Ph.D.Instituto Biología Vegetal y Biotecnología Universidad de Talca2 Norte 685phone 56-71-200280fax 56-71-200276

DAAD funded researcher exchange15. September 2010 – 20. May 2011

Miriam E González Rojas; PhD studentInstituto de Biología Vegetal y BiotecnologíaUniversidad de TalcaCasilla 747, Talca, ChileFono: +56-71-200275Fax: +56-71-200276

On-going collaboration with Chilean researchers

Project:Biochemical characterization of FcAAT1 an enzyme involved in the formation of fruit esters in strawberry

FcAAT1



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.)


Strawberry Bet v 1-homologous allergen (Fra a 1) is down-regulated in colorless strawberry mutant

Hjernø et al., Proteomics 2006, 6, 1574-1587



Functional genomics in fruit: Transient overexpression or RNAi-mediated gene silencing in ripening strawberry fruit

A B C

D FE14 days

chalcone synthase (CHS)

Agroinfiltration

Hoffmann et al. Plant J. 2006, 48, 818


Intron


Nucleotides alignment

Proteins alignment

m109b07R Fra a 1e

c201c06R Fra a 2

m117g04R Fra a 3

Fra a 1 Homologue of Bet v1 in strawberry

Munoz et al., Molecular Plant 2010, 3, 113



Fra a 1

Fra a 2

Fra a 3

RT-PCR reactions.

Were performed with primers specific for the ESTs M109B07, C201C06 or M117G04.

The interspacer gene was used as control.

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

RT-PCR analysis of Fra a 1, Fra a 2 and Fra a 3 in strawberry



Downregulation of the strawberry allergen Fra a

Negative control (pBI-intron)

Positive control (CHSi)

Fra a 3

Fra a 1



Phe

coumaroyl CoA

coniferyl alcohol

lignin phenylpropenoidsbenzenoids

flavonoids

anthocyanins

PAL

CHS

Fra a

allergen

color firmness flavor

Effects of Fra a

agronomic traits

pathogenesis-related (PR)

protein

resistance




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

?


O

O

HO OH

OPO3--OH--O3PO

OHO

O

OHOFaQR

O

OH3CO

FaOMT glucosyl transferase

OHO

HOOH

OH

O

OO

OHO

HOOH

HOOC-CH2-OOC

O

OO

malonyl transferase

D-fructose-1,6-diphosphate

HDMF

HDMF glucosideDMMF

HDMF glucoside malonate

HMMF

O

OHHO

O

OOketo-enoltautomerism

keto-enoltautomerism

Biosynthesis of the key strawberry flavor compound (HDMF)

Wein et al., Plant J., 2002, 31, 755Lavid et al., J. Agric. Food Chem., 2002, 50, 4025Lunkenbein et al., J. Exp. Bot., 2006, 57, 4025

Raab et al., Plant Cell, 2006, 18, 1023Klein et al., J. Agric. Food Chem., 2007, 55, 6705



Biotechnological production of norisoprenoids










4 biotechnological production of enzymes used for food industry (alpha-amylase, glucose isomerase, chymosin, pectinase, lipase, lipoxygenase, dioxygenases)



C9 Volatile Products

9-LOX13-LOX

9-HPL

13-HPL

‘Green note‘ production

Hexanal, Hexenals

Nonenals, Nonadienals



Transient expression of LOX and HPL genes


FlavorFaOMT, FaQR furaneol (strawberry)FaAAT1, FcAAT fruit esterFaCCD1, RdCCD1 norisoprenoid, eg. -iononeFaNES, FaPIN, LaLIN, LaBER, LaLIM mono- and sesquiterpenesFaEGS phenylpropenes, eg. eugenolFaJMT methyl jasmonateCrSMT S-methyltransferase thiomethyletherTbLOX fatty acid hydroperoxide aldehydes PigmentFaGT1 anthocyan FaGT6, 7 flavonoidFaCHS, FaANS, FaDFR, FaANR anthocyan

Firmness shelf lifeFaCCR, FaCAD, FaPOD lignin

Pathogene resistance allergenicityFra a 1,2,3 affects anthocyan formation, allergensFaGT2, 5 hydroxycinnamoyl glucose

Proteins related to food industry



Phytoenzymes

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.

plants are underrepresented as source for industrial biocatalysts:

Towards functionalized natural products through exploitation of the plant gene pool

newly sequencedplant genomes

A. GierlW. Schwab

substrates/products,biochemical function

cloned enzyme libraries

biological context,structural analysis,protein engineering

T. Rattei

T. HofmannB. Küster

A. SkerraW. Liebl

target sequencesmodels

A. Schiefner I. Antes

newly sequencedplant genomes

A. GierlW. Schwab

substrates/products,biochemical function

cloned enzyme libraries

biological context,structural analysis,protein engineering

T. Rattei

T. HofmannB. Küster

A. SkerraW. Liebl

target sequencesmodels

A. Schiefner I. Antes

dioxygenases



and DFG, DAAD, Firmenich, Degussa, BMBF, EU, AIF, ZuTechfor financial support

THANKS TO


technische universität münchen prof. dr. wilfried schwab improving food production by...

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food production

food products

food sciences plant

food quality

production of food bread

value food crops biotechnology

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