8/05/2017. Zuurdesemproductie: backslopping of gestuurde fermentatie? Contents. Contents. Introduction. Classification. Impact of process conditions

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1 Zuurdesemproductie: backslopping of gestuurde fermentatie? ir. Simon Van Kerrebroeck Studiereeks Micro-Organismen Part 1 - The Good & The Ugly Brewery De Koninck, Antwerp May 11, 2017 Research Group of Industrial Microbiology and Food Biotechnology Prof. Dr. ir. Luc De Vuyst Prof. Dr. ir. Frédéric Leroy Prof. Dr. Stefan Weckx Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 2 Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 3 1

2 Introduction Sourdough Sourdough is a mixture of cereal fractions (flour) and water that is fermented to a low ph by a microbial ecosystem containing lactic acid bacteria (LAB) and yeasts. Sourdough can be used for the production of baked goods 4 Introduction Technology Dough processing Bread texture Flavor Shelf-life Gluten Consumer Artisan, natural, gastronomical status Nutritional aspects (gluten) Renewed interest Over eten, VRT 5 Introduction > Lactic acid bacteria (LAB) and yeasts Specific and stressful ecosystem Low ph Carbohydrates to be shared Low oxygen tension Higher cell counts of LAB (> 10 8 CFU/g) compared to yeasts (< 10 7 CFU/g) Major metabolic activities Acidification (LAB) Flavor formation (LAB, yeasts) Leavening (yeasts, LAB) Corsetti & Settanni (2007) FRI 40 6 De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM 100 2

3 Introduction The sourdough microbiota LAB Yeasts % of sourdoughs 47 Lactobacillus sanfranciscensis Lactobacillus brevis 14 Pediococcus pentosaceus 68 Saccharomyces cerevisiae 20 Kazachstania/Candida humilis Pichia kudriavzevii 6 Torulaspora delbrueckii Wickerhamomyces anomalus 13 Lactobacillus paralimentarius 5 Candida glabrata Based on a meta-analysis of 583 backslopped sourdoughs 7 De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM 100 Introduction Origin of the sourdough microbiota House microbiota Agricultural practices, weather (rainfall), etc. Textile and materials Baker s yeast Fruits, berries, leaves Lactobacillus sanfranciscensis Diverse yeast species 8 From grain field, over storage, to bakery Intestinal origin (faeces) Lactobacillus reuteri De Vuyst et al., 2014, FM 37; 2017, AAM 100; Ripari et al., IJFM 232; Gobbetti et al., 2016, FM 60; Minervini et al., 2012, AEM 78; 2015, AEM 81; FM 52; 2016, FM 60; Scheirlinck et al., 2009, JAM 106 Introduction Interactions Process conditions Temperature Time Dough Yield (DY) Storage Day/night, seasonal Ingredients Microbiota Enzymes Amylases Carbohydrate release Proteases Supply peptides to LAB and yeasts Lipooxygenases, phytases Species diversity Community dynamics Metabolite kinetics Process conditions ph Redox balance Gradients 9 De Vuyst et al., 2014, FM 37; 2017, AAM 100; Gänzle, 2014, FM 37 3

4 Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 10 Classification According to region According to inoculation type Type 1 Type 2 Type 3 According to process conditions Type 0 Type I Type II Type III De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM Classification Region Examined in a.o. Belgium, China, France, Italy Could not be confirmed in a meta-analysis Research reflects regionally important cereals Link with process factors and environment? Certain yeasts associated with use of textile cloth or fruits Lactic acid bacteria (Lactobacillus sanfranciscensis) from berries and fruits Leuconostoc citreum from milk Rye production/yield Wheat production/yield AndrewMT, 2010/Monfreda et al.,2008, GBC 22 Lattanzi et al., 2013, IJFM 163; Lhomme et al., 2015, IJFM 215; Minervini et al., 2012, AEM 78; Zhang et al., 2011, FC De Vuyst et al., 2017, AAM 100; Scheirlinck et al., 2007, AEM 73 4

5 Classification Process conditions Type 0: Pre-doughs/sponge doughs: saves the use of baker s yeast, improves taste and flavor Type I: Traditional, firm bakery sourdoughs: continuously propagated; C; DY < 200; ph ~4.0; acidification, flavoring, and leavening Type II: Industrial, liquid sourdoughs with the addition of a starter culture: prolonged fermentation time; C; DY > 200; ph ~3.5; dough acidification, improving and flavoring; requires addition of baker s yeast because of natural yeast inhibition by LAB acidification Type III Dried preparations of industrial sourdoughs 13 De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM 100 Classification Process conditions Type I-II influenced the number of LAB/yeast species in backslopped sourdoughs Number of LAB/yeast species Type I LAB Type II Type yeasts I Type II 14 De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM 100 Classification Process conditions Type I-II influenced the number of LAB/yeast species in backslopped sourdoughs Influence of Lactobacillus sanfranciscensis Type I Number of LAB/yeast species Type Type I - I LAB + Type Type II II Type Type I yeasts I - + Type Type II II 15 De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM 100 5

6 Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 16 Periodic refreshment of the dough by re-inoculation of a fresh flourwater mixture with fermented dough Vogelmann & Hertel, 2011, FM De Vuyst et al., 2014, FM Selection towards the most adapted microorganisms based on rebuilding the microbial communities Lactobacillus sanfranciscensis: small genome, matrix-adapted (e.g. maltose-positive, auxotrophy for amino acids, amino acid conversion) Lactobacillus reuteri: glycerol metabolism or on maintaining the microbial population Lactobacillus reuteri: glutamate decarboxylase pathway contributes to ph tolerance Lactobacillus fermentum: arginine deiminase pathway 18 Lin & Gänzle, 2014, AEM 80 ; Vrancken et al., 2008, IJFM 128; 2009, IJFM 135, FM 26 6

7 Lactic acid bacteria metabolism 19 De Vuyst et al., 2014, FM 37; 2016, IJFM 239; 2017, AAM 100 Selection towards the most adapted microorganisms Day 0-3 Lactococcus Enterococcus Leuconostoc Day 3-5 Weissella Pediococcus Lactobacillus Day 5-10 Lactobacillus fermentum 20 Van der Meulen et al., 2007, 73; Weckx et al., 2010, AEM 27, FM 76; Harth et al., 2016, IJFM 228; Van Kerrebroeck et al., 2016, IJFM 239 Selection towards the most adapted microorganisms Temperature Influences species diversity Time Longer fermentation exhausts nutrients and prolongs exposure to stress-conditions (ph) Vogelmann & Hertel, 2011, FM De Vuyst et al., 2014, FM 7

8 30 C 24 h Lactobacillus fermentum 23 C 24 h Leuconostoc citreum 37 C 24 h Lactobacillus fermentum 30 C 48 h Lactobacillus fermentum Lactobacillus plantarum Laboratory sourdough, wheat, 10 backsloppings 22 Vrancken et al., 2011, AEM 77 Selection towards the most adapted microorganisms Temperature Influences species diversity Time Longer fermentation exhausts nutrients and prolongs exposure to stress-conditions (ph) ph Influences microbial community dynamics and metabolite production kinetics Vogelmann & Hertel, 2011, FM De Vuyst et al., 2014, FM a Laboratory sourdoughs Temperature of 30 C Dough yield of 400 (liquid dough) Constant stirring Daily backslopping b Free ph ph > 4.0 (ph-capped with 5 M NaOH) 24 Van Kerrebroeck et al., 2016, IJFM 239 8

9 a (Lb. fermentum) Lb. plantarum Kazachstania sp. Lb. fermentum (Lb. plantarum) Candida glabrata b Continuous ethanol production Continuous presence of glucose Higher lactic acid concentrations Free ph ph > 4.0 (ph-capped with 5 M NaOH) 25 Van Kerrebroeck et al., 2016, IJFM 239 Selection towards the most adapted microorganisms Temperature Influences species diversity Time Longer fermentation exhausts nutrients and prolongs exposure to stress-conditions (ph) ph Influences microbial community dynamics and metabolite production kinetics Dough yield Vogelmann & Hertel, 2011, FM De Vuyst et al., 2014, FM Selection towards the most adapted microorganisms Temperature Influences species diversity Time Longer fermentation exhausts nutrients and prolongs exposure to stress-conditions (ph) ph Influences microbial community dynamics and metabolite production kinetics Dough yield Provides buffer capacity and nutrients, increases the effect of the flour Vogelmann & Hertel, 2011, FM De Vuyst et al., 2014, FM 9

10 Laboratory DY 400 T 30 C Stirring Daily refreshment Lb. brevis Lb. fermentum Lb. plantarum Bakery DY 200 T ~ 20 C No stirring Daily refreshment Weissellas Leuconostocs Wider and different microbial diversity, slower evolution to final microbial communities 28 Harth et al., 2016, IJFM 228 Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 29 Impact of cereals Hits on Web of Science, period Wheat 685 hits Rye 205 hits Barley 46 hits Oat 35 hits Spelt 11 hits Teff 10 hits References with microbial analysis on sourdough, period ref. 22 ref. 3 ref. 1 ref. 7 ref. 5 ref

11 Impact of cereals Is there a connection between cereal and microbial diversity? Wheat, rye, barley, and spelt examined under same laboratory conditions Temperature of 30 C Dough yield of 400 (liquid dough) Constant stirring Daily backslopping Microbiological analysis of the lactic acid bacterial communities after 10 days 31 Van der Meulen et al., 2007, 73; Weckx et al., 2010, AEM 27, FM 76; Harth et al., 2016, IJFM 228 Impact of cereals Wheat Lb. fermentum Rye Lb. plantarum Lb brevis P. pentosaceus Barley Spelt 32 Van der Meulen et al., 2007, 73; Weckx et al., 2010, AEM 27, FM 76; Harth et al., 2016, IJFM 228 Impact of cereals Wheat Lb. fermentum Rye P. pentosaceus Lb. plantarum Lb. brevis Barley Spelt Lb. rossiae 33 Van der Meulen et al., 2007, 73; Weckx et al., 2010, AEM 27, FM 76; Harth et al., 2016, IJFM

12 Impact of cereals C4-plants Millets, sorghum, maize, teff Lactobacillus sanfranciscensis less competitive No maltose, but sucrose/glucose High contents of phenolic compounds Buckwheat and oats limit yeast growth? Reasons not known A, wheat; C, sorghum Sekwati-Monang et al., 2012, IJFM 159 Vogelmann et al., 2009, IJFM Harth, 2016, unpublished results Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 35 New developments: health, additives, and taste Gluten Use of sourdough based on certain flours for gluten-free products o From 2 publications < 2000 to 80 in o Baking quality Enzymatic degradation by fungal proteases and lactic acid bacteria (Bioactive) peptides and amino acids ACE-inhibitory (anti-hypertension), anti-cancer (lunasin), anti-oxidant, mood-enhancing (GABA) Taste effect (salt replacement) glutamate (umami, savory) γ-glutamyl dipeptides (kokumi, taste-enhancing) 36 Nionelli et al., 2016, Foods 5; Rizzello et al., 2007, AEM 73; Zhao & Gänzle, 2016, JAFC 64 12

13 New developments: health, additives, and taste Antifungal activity Acetic and propionic acid Other carboxylic acids (phenyllactic acids) Texture o Exopolysaccharides o Organic acids Removal of undesired compounds Phytate: anti-nutritional factor Vicine: faba bean flour Phenolic compounds Use of bran, wheat germ, other seeds (chestnuts, sunflower, etc.) Application of novel starter cultures, in particular non-conventional yeasts and bacteria Alvarez-Jubete, 2010, TFST 27; Arendt et al., 2007, FM 24; Axel et al., 2016, AMB 100; Nionelli et al., 2016, Foods 5; Rizzello et al., 2007, AEM 73; Zhao & Gänzle, 2016, JAFC De Vuyst et al., 2017, AAM 100 Contents Introduction Classification Impact of cereals New developments: health, additives, and taste Conclusions 38 Conclusions Different origins for sourdough microbiota, but Process factors are largely determining the microbial species diversity Flour can have an impact too Region specificity could not be retained New developments focus on health (gluten), ingredient/additive replacement (antifungal, texture, salt), and taste by testing novel starter cultures (bacteria and yeasts), new or ancient cereals, and improved process understanding 39 13

14 Acknowledgments Prof. Dr. ir. Luc De Vuyst Prof. Dr. ir. Frédéric Leroy Prof. Dr. Stefan Weckx Dr. MSc. Henning Harth MSc. Andrea Comasio ir. Natalia Savelieva IMDO team SRP, IRP, IOF projects 40 Subdivision into types Temperature, fermentation time Dough yield below 200 Dough yield above 200 Temperature up to 30 C n = 85 Fermentation time below ph above h Lactobacillus sanfranciscensis Weissellas-Leuconostocs Candida humilis Saccharomyces cerevisiae Temperature up to 30 C Fermentation time above 12 h Temperature above 30 C n = 89 ph above or below 3.8 Lactobacillus sanfranciscensis Saccharomyces cerevisiae Kazachstania spp. n = 14 Lactobacillus delbrueckii-group No yeasts-pichia kudriavzevii n = 10 ph mainly above 3.8 Leuconostocs Kazachstania spp. Saccharomyces spp. n = 26 ph mainly below 3.8 Diverse yeasts species n = 14 Lactobacillus delbrueckii-group Diverse yeasts species 41 Sourdough Influence process conditions Temperature, fermentation time Dough yield below 200 Dough yield above 200 Temperature below 30 C n = 85 Fermentation time below ph above h Lactobacillus sanfranciscensis Weissellas-Leuconostocs Candida humilis Saccharomyces cerevisiae Temperature below 30 C Fermentation time above 12 h Temperature above 30 C n = 89 ph above or below 3.8 Lactobacillus sanfranciscensis Saccharomyces cerevisiae Kazachstania spp. n = 14 Lactobacillus delbrueckii-group No yeasts-pichia kudriavzevii n = 10 ph mainly above 3.8 Leuconostocs Kazachstania spp. Saccharomyces spp. n = 26 ph mainly below 3.8 Diverse yeasts species n = 14 Lactobacillus delbrueckii-group Diverse yeasts species 42 14