Fermenterede grøntsager

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1 Fermenterede grøntsager Hvad er fermentering Egon Bech Hansen

2 Min egen baggrund Civilingeniør og PhD fra DTU Forskningsdirektør for Chr Hansen A/S Forskningsdirektør for Kulturdivisionen i Danisco A/S Professor i indistriel bioteknologi, Aalborg Universitet Professor i mikrobielle fødevareingredienser, DTU Food

3 Agenda Fermentering, historie og formål Metabolisme Hvilke mikroorganismer anvendes Sikkerhed og kontrol Podning og podemateriale

4 Fermentering 1. Anaerob mikrobiologisk proces ( la vie sans air, Pasteur) 2. Mikrobiologisk proces 3. Proces med mikroorganismer og enzymer

5 Et århundrede med fermentering Sigurd Orla Jensen Professor in Biotechnical Chemistry Papers still often cited: S Orla-Jensen (1942): The lactic acid bacteria S Orla-Jensen (1909): Die Hauptlinien des natürlichen Bakteriensystems. Zentbl. Bakteriol. Parasitenkd. Infektkrank. Hyg. Abt. S Orla-Jensen (1921): The main lines of the natural bacterial system. Journal of bacteriology S Orla-Jensen and J. & A. Churchill (1921): Dairy bacteriology S Orla-Jensen, E Olsen, T Geill (1949): Senility and Intestinal Flora A Reexamination of Metchnikoffs Hypothesis Journal of gerontology

6 Velkendte eksempler på fermenterede fødevarer Kærnemælk Ost Øl Vin Eddike Surkål Brød hævet med surdej Brød hævet med gær Soyasauce Fermenterede pølser (marinerede sild)

7 Formål med fermentering Konservering Organiske syrer: mælkesyre, eddikesyre, propionsyre, etc. Produktion af alkohol Produktion af antimikrobielle stoffer, bakteriociner, diacetyl, etc. Kompetitiv eksklusion Smag og aroma Forbrugerne har udviklet smagspræferencer for fermenterede produkter Fjerne giftstoffer og antinæringsstoffer Helsebringende virkning (Siden: Metchnikoff, E., The Prolongation of Life)

8 Fokus på helse Human microbiome 18 species in all 57 species in >90% 75 species in >50% Large common core Minimal human gut metagenome encompasses functions important for the host bacterial interaction Degradation of complex polysaccharides Synthesis of short chain fatty acids, indispensable amino acids, vitamins Relative abundance of 57 frequent microbial genomes JJ Qin et al. Nature 464, (2010)

9 Fokus på helse Probiotika

10 Fokus på helse

11 Fokus på helse

12 De metaboliske processer med eller uden luft? Aerob metabolisme Effektiv produktion af energi Bredt spektrum af mulige processer Tilstedevær af ilt kan afbalancere redoxprocesser Anaerob metabolisme (fermentering sensu stricto) De metaboliske processer skal indbyrdes afbalancere redoxreaktionerne Kun få energigivende reaktioner Stor rigiditet i spektret af mulige processer Næringsværdi bevares bedst ved anaerobe processer Produktion af store mængder af væsentlig ændret biomasse vil ofte kræve aerobe processer

13 De metaboliske processer

14 The main metabolic pathways for production of lactic acid by lactic acid bacteria

15 Pyruvate metabolism of Lactococcus lactis C. M. Henriksen and D. Nilsson Appl Microbiol Biotechnol (2001) 56:

16 Different sugars different routes

17 Hvilke mikroorganismer anvendes? Veldefinerede kulturer eller komplekse mikrobielle konsortier? Product Raw material Starter Culture Beer Cereals Yeast Wine Grape juice Yeast, lactic acid bacteria Bread Grains Yeast, lactic acid bacteria Soy sauce Soybeans Mould (Aspergillus), lactic acid bacteria Sauerkraut, Kimchi Cabbage Lactic acid bacteria Fermented Sausages Meat Lactic acid bacteria Pickled vegetables Cucumbers, olives a.o. Lactic acid bacteria Fermented milks Milk Lactic acid bacteria Cheese Milk Lactic acid bacteria, yeast, mould

18 Regulatory systems for Microbial Food Cultures USA Food Drug and Cosmetic Act (1958) introduced the status of Generally Recognized As Safe (GRAS) European Union EFSA Qualified Presumption of Safety (QPS) Denmark National legislation since 1974

19 History of Use Proposed definition according to Health Canada, 2003 Significant human consumption of food over several generations and in a large, genetically diverse population for which adequate toxicological and allergenicity data exists to provide reasonable certainty that no harm will result from consumption of the food

20 Microbial Taxonomy The definition of species of microorganisms lacks a theoretical basis In praxis a species is represented by a type strain Strains showing a high degree of phenotypic and/or genotypic similarity to that type strain are regarded as belonging to the same species Objective measures of relatedness have been proposed, but there is no simple definition of the species as a taxonomical unit Fungi are allowed to have two names one for teleomorph and holomorph state one for the anamorphic state

21 Undesired properties of MFC Infections and opportunistic infections Toxic metabolites Virulence factors Antibiotic resistance

22 C. botulinum C. tyrobutyricum B. casei K. varians B. linens M. tuberculosis P. acidipropionici P. freudenreichii O. oeni B. lactis S. pneumoniae S. thermophilus Lactococcus lactis W. halotolerans W. confusa L. pseudomesenteroides L. mesenteroides L. lactis E. faecium E. faecalis C. divergens C. piscicola L. monocytogenes S. aureus S. carnosus S. xylosus B. cereus L. delbrueckii L. acidophilus L. panis L. fermentuml. pentosus L. gasseri L. reuteri L. plantarum L. johnsonii L. brevis L. casei L. rhamnosus L. sakei L. alimentarius L. sanfranciscen. L. kefiri P. damnosus P. pentosaceus P. acidilactici E.coli B. bifidum B. adolescentis B. infantis B. longum A. aceti 0.1

23 Modern Science and Ancient Microorganisms Up-to-date inventory of microbial species used in production of fermented foods The inventory covers species of starter cultures and "natural floras History of use also for newly established taxonomic units The inventory consists of 185 bacterial species and 36 species of yeasts and moulds Bourdichon, F. et al., Food fermentations: Microorganisms with technological beneficial use. International Journal of Food Microbiology, 154(3), pp

24 Bacterial Diversity in the 2011 list

25 Fungal diversity in the 2011 list

26 Microorganisms used in dairy fermentations Bacteria Fungi Arthrobacter arilaitensis Enterococcus durans Lactobacillus rhamnosus Candida colliculosa Arthrobacter bergerei Enterococcus faecalis Lactobacillus salivarius Cystofilobasidium infirmominiatum Arthrobacter globiformis Enterococcus faecium Lactobacillus tucceti Debaryomyces hansenii Arthrobacter nicotianae Hafnia alvei Lactococcus lactis Fusarium domesticum Bifidobacterium adolescentis Kocuria rhizophila Lactococcus raffinolactis Geotrichum candidum Bifidobacterium animalis Kocuria varians Leuconostoc citreum Issatchenkia orientalis Bifidobacterium bifidum Lactobacillus acidipiscis Leuconostoc mesenteroides Kazachstania exigua Bifidobacterium breve Lactobacillus acidophilus Leuconostoc pseudomesenteroides Kazachstania unispora Bifidobacterium longum Lactobacillus brevis Macrococcus caseolyticus Kluveromyces marxianus Bifidobacterium pseudolongum Lactobacillus casei Microbacterium gubbeenense Kluyveromyces lactis Bifidobacterium thermophilum Lactobacillus coryniformis Micrococcus luteus Lecanicillium lecanii Brachybacterium alimentarium Lactobacillus delbrueckii Propionibacterium acidipropionici Mucor plumbeus Brachybacterium tyrofermentans Lactobacillus helveticus Propionibacterium freudenreichii Mucor racemosus Brevibacterium aurantiacum Lactobacillus kefiri Propionibacterium jensenii Penicillium camemberti Brevibacterium casei Lactobacillus kefranofaciens Propionibacterium thoenii Penicillium caseifulvum Brevibacterium linens Lactobacillus nodensis Staphylococcus equorum Penicillium chrysogenum Carnobacterium divergens Lactobacillus parabrevis Staphylococcus fleurettii Penicillium commune Carnobacterium maltaromaticum Lactobacillus paracasei Staphylococcus sciuri Penicillium nalgiovense Corynebacterium ammoniagenes Lactobacillus parakefiri Staphylococcus succinus Penicillium roqueforti Corynebacterium casei Lactobacillus pentosus Staphylococcus vitulinus Pichia fermentans Corynebacterium flavescens Lactobacillus perolens Staphylococcus xylosus Yarrowia lipolytica Corynebacterium variabile Lactobacillus plantarum Streptococcus gallolyticus Zygotorulaspora florentina Streptococcus salivarius

27 Microorganisms used in cheese production Bacteria Fungi Arthrobacter arilaitensis Lactobacillus pentosus Candida colliculosa Arthrobacter bergerei Lactobacillus perolens Cystofilobasidium infirmominiatum Arthrobacter globiformis Lactococcus raffinolactis Debaryomyces hansenii Arthrobacter nicotianae Leuconostoc citreum Fusarium domesticum Brachybacterium alimentarium Macrococcus caseolyticus Geotrichum candidum Brachybacterium tyrofermentans Microbacterium gubbeenense Kluveromyces marxianus Brevibacterium aurantiacum Micrococcus luteus Kluyveromyces lactis Brevibacterium linens Propionibacterium acidipropionici Lecanicillium lecanii Corynebacterium ammoniagenes Propionibacterium freudenreichii Mucor plumbeus Corynebacterium casei Propionibacterium jensenii Mucor racemosus Corynebacterium flavescens Propionibacterium thoenii Penicillium camemberti Corynebacterium variabile Staphylococcus equorum Penicillium caseifulvum Enterococcus faecalis Staphylococcus fleurettii Penicillium chrysogenum Enterococcus faecium Staphylococcus sciuri Penicillium commune Kocuria rhizophila Staphylococcus vitulinus Penicillium nalgiovense Lactobacillus coryniformis Penicillium roqueforti Lactobacillus parabrevis Yarrowia lipolytica

28 Microorganisms used in meat fermentations Species Food Usage Species Food Usage Carnobacterium divergens Meat, Fish, Cheese Micrococcus lylae Carnobacterium piscicola Meat Pediococcus acidilactici Meat Halomonas elongata Meat Pediococcus pentosaceus Meat Kocuria rhizophila Cheese Meat Penicillium solitum Meat Kocuria varians Dairy, Meat Staphylococcus carnosus Meat Lactobacillus alimentarius Fish, Meat Staphylococcus equorum Cheese, meat Lactobacillus curvatus Meat Staphylococcus succinus Meat Lactobacillus dextrinicus Meat Staphylococcus vitulinus Cheese, meat Lactobacillus paracasei Dairy, Meat Staphylococcus warneri Meat Meat Lactobacillus rhamnosus Dairy, Vegetables, Meat Steptomyces griseus Meat Lactobacillus sakei Meat, Sake Weissella paramesenteroides Meat Leuconostoc carnosum Meat

29 Soy fermentations Species Aspergillus oryzae Aspergillus sojae Bacillus subtilis Enterococcus faecalis Enterococcus faecium Lactobacillus farciminis Mucor hiemalis Rhizopus oligosporus Staphylococcus condimenti Tetragenococcus halophilus Food Usage Japanese sake, Soy sauce, Miso Soy sauce, Miso Natto, soybean proteolysis Ham, Cheese, Miso and Japanese pickles fermentation Ham, Cheese, Miso and Japanese pickles fermentation Fish, Soy Fermented soy bean curd Tempeh Soy Soy sauce, Miso

30 Kimchi Species Tetragenococcus koreensis Lactobacillus kimchii Leuconostoc inhae Leuconostoc kimchii Weissella cibaria Weissella koreensis Food Usage Kimchi Kimchi Kimchi Kimchi Kimchi Kimchi

31 IITA Image Libr Cassava fermentation Species Weissella beninensis Food Usage Cassava fermentation

32 Gram negative bacteria used in food fermentation Species Acetobacter aceti Food Usage Vinegar Species Gluconacetobacter diazotrophicus Food Usage Cocoa, Coffee Acetobacter fabarum Cocoa, Coffee Gluconacetobacter entanii Vinegar Acetobacter lovaniensis Vegetables Gluconacetobacter europaeus Vinegar Acetobacter malorum Vinegar Gluconacetobacter hansenii Vinegar Acetobacter orientalis Vegetables Gluconacetobacter johannae Cocoa, Coffee Acetobacter pasteurianus Vinegar, Cocoa Gluconacetobacter oboediens Vinegar Acetobacter pomorum Vinegar Gluconacetobacter xylinus Vinegar Acetobacter syzygii Vinegar, Cocoa Hafnia alvei Dairy Acetobacter tropicalis Cocoa, Coffee Halomonas elongata Meat Gluconacetobacter azotocaptans Cocoa, Coffee Zymomonas mobilis Beverage

33 Hvordan startes en fermentering? Begrænsning af uønskede mikroorganismer Friske og sunde råvarer Undgå uønskede tilblandinger Evt tilsættes salt, sukker eller andre ingredienser der begrænser vækst (traditionel hurdle teknologi) Evt varmebehandling Evt begrænses adgang af luft Podning Spontan fermentering Back slopping Podning med starterkultur

34 Komercielle starterkulturer størrelse af markedet fordelt på typer Distribution of world market on culture types Meat Probiotics Fermented Milks Cheese Total value of the global culture market: mio

35 Microorganisms produced in largest volumes 1. Saccharomyces cereviciae Bakers yeast (~ 1 million t) Brewers yeast Inoculum for wine making 2. Lactococcus lactis (~ t) cheese, fermented milks bread, meat, vegetables probiotics, bioprotection 3. Streptococcus thermophilus ( t) cheese, fermented milks bread, meat, vegetables probiotics

36 Inoculum for nye typer af fermenterede produkter 1. Spontan fermentering 2. Back slopping med tidligere produkter som opfylder forventning til smag, kvalitet og sikkerhed 3. Udvikling til egentlige starterkulturer

37 Et eksempel fra Etiopien

38 Haramaya Camel Dairy Project Danida funded project with 8.9 mio DKK 5 years from Jan 2013 to Dec 2017 Develop dairy products based on camel milk 2 PhD and 10 MSc projects

39 Top 10 bacterial genera in traditional Ethiopian fermented camel milk _Streptococcus _Weissella _Streptococcus _Escherichia _Lactococcus _Klebsiella _Klebsiella _Clostridium

40 Tak for jeres opmærksomhed og interesse