inauguration sourdough library · 2019-02-18 · kneading (20 min) portioning and shaping 18-25°c,...

Inauguration Sourdough Library

15 October 2013

Professor of Microbiology at the Department of Soil, Plant and Food Sciences University of Bari, Italy

“History of bread”

Marco Gobbetti

16 – 17 October, Sankt Vith (Belgium)

Department of Soil, Plant and Food Science, University of Bari Aldo Moro, Italy

Bread: «The ferment of life»

Wall painting of the Tomb of Ramesses III, (1570-1070 b. C), XIX Dinasty


For the Egyptians: Sacred value….”a gift of God or the gods”

For the Egyptians: a piece of merchandise..

For the Christians..Eucharist

For the Jews.. sacred and transcendent value

For the Latin.. vehicle for transmitting of the sacrum

For the Greeks..offered to the Divinity…medicinal purpose

For the Romans..sign of purification

Influence of the term «bread» on the common lexicon


«Companion» (from cum panis)

“man cannot live on bread alone” “to eat unearned bread”

«Lord» (from Old English vocabulary hlaford)

“remove bread from his mouth”

“to earn his bread”

History and sourdough

Egyptians (2000 B.C.) casually discovered a leavened dough; used foam of beer as a (unconscious) starter for dough leavening

Romans (1° century A.D.) used to propagate the sourdough through back-slopping (Plinio il Vecchio, Naturalis Historia XVIII)

Middle Age (1600): dawn of the use of baker’s yeast for bread


(FromTheatrum sanitatis, XI Century)

Tacuini sanitatis (XI century): “among the six elements needed to keep daily wellness… foods and beverages… “ “…White bread: it improves the wellness but it must be completely fermented …“



Pliny the elder wrote: “….Then, normally they do not even heat the dough, but they just use a bit of dough left from the day before, and it is undeniable that flour, by its nature, is leavened by an acid substance. …“(Pliny the Elder G (1972) Naturalis Historia XVIII, 102-104, edition of Le Biniec H.

History and sourdough

2013

Egyptian (2000 B.C.) casually discovered a leavened dough; used foam of beer as a (unconscious) starter for dough leavening

Romans (1° century A.D.) used to propagate the sourdough through back-slopping (Plinio il Vecchio, Naturalis Historia XVIII)

Middle Age (1600): dawn of the use of baker’s yeast for bread

1900: sourdough being more and more replaced by baker’s yeast

discovery of the sourdough (consumer / industry)


1800: Louis Pasteur found the agents of fermentation


ASCB, Comune, serie terza, sec. XIX, mazzo 62 fascicolo 031 - 031.01

Bread as an invaluable good...


Celebration of bread, 1928 Milano, Italy (Civic Collection of prints “Achille Bertarelli”)

Bread is good for all...

And finally came the sweet leavened products





Nowadays...cultural heritage


What is the sourdough?

“A mixture of flour (wheat, rye, rice, etc…) and water, fermented by lactic acid bacteria

and yeasts, which are responsible for its capacity to leaven a dough, while

contemporarily and unavoidably acidifying it”

(Gobbetti, 1998. Trends Food Sci. Technol.)


How may lactic acid bacteria contribute to sourdough processes?

Glucose

Piruvate (2) Lactate

NADH+H+ NAD

Fructose

Glucose Sucrose Maltose

Maltose Glucose

H + H +

+ +

6P-Gluconate NAD + NADH + H + NAD + NADH + H + CO2

Acetyl-P P i

3P-Glyceraldehyde

5P-Ribulose

P i 5P-Xilulose

Acetate ADP ATP

ADP ATP

Ethanol Piruvate

NADH+H

NAD

(1) Lactate

Heterolactic fermentation Homolactic fermentation

alkaline

protein

oligopeptides

PrtP

ATP amino acids

biosynthesis

H+

amino acids

Moderate increase of the volume

Synthesis of volatile compounds

Acidification

Acidification

CO2

Increase of the volume


How may yeasts contribute to sourdough processes?

Synthesis of volatile compounds

Acetaldehyde

Ethanol

Fatty acids Esters Alcohols

H2S

ATP

Diidrissiacetonfosfato 3 - P Glyceraldehyde TPI

FBA

Acetaldehyde

Pyruvate

Ethanol Ethanol

Ethanol

Carbohydrates

Carbohydrates

Acetyl-CoA

Fatty acids CoA

Fatty acids Esters

H2S Suphates/sulfites

Amino acids

Keto acids

Higher alcohols

Diacetyl

Acetolactate


Where are from lactic acid bacteria and yeasts?

Where are from lactic acid bacteria and yeasts?

Ecological parameters

ü  Dough yield ü  Sourdough (%) used as

the inoculum ü  NaCl ü  Redox potential ü  Fermentation time ü  F e r m e n t a t i o n

temperature ü  pH ü  N u m b e r o f b a c k -

slopping step ü  Storage temperature

(Minervini et al., 2013. Int. J. Food Microbiol.)


How to prepare the sourdough?


Rye

Triticum durum

T. aestivum

(Ercolini et al., 2013. Appl. Environ. Microbiol.)

0 1 2 5 10

Days of propagation (refreshment)

Daily sourdough back slopping Sampling


Sourdough: “a mixture of flour and water, fermented by lactic acid bacteria and

yeasts, which are responsible for its capacity to leaven a dough” (Gobbetti et al., 1998)

Type I sourdough. Traditional sourdoughs whose microrganisms are kept metabolically active through daily refreshments (three-stage protocol is applied relying on three refreshments over 24 h).

Classification of sourdoughs


Lactobacillus sanfranciscensis L. pontis, L. fructivorans L. fermentum, L. brevis

Candida milleri, C. holmii, Saccharomyces cerevisiae, Kazachstania exigua

+

Sourdough

Water Flour 1° refreshment – fermentation at ca. 25°C for 5 - 6 h

Leavening agent for bread production

2° refreshment – fermentation at ca. 28°C for 7 - 8 h

3° refreshment – fermentation at ca. 25°C for 2 - 3 h

Water

Water

Flour

Flour

Mature sourdough

Mother sponge stored till using for the next bread-making

Type II sourdough. Sourdoughs obtained through a unique fermentation step of 15 – 20 h followed by storage for many days. These sourdoughs are generally liquid (DY of ca. 200) and they are produced at the industrial level using bioreactors or tanks at a controlled temperature that exceeds 30°. Type II sourdoughs are used for dough acidification, and as dough improvers.

Lactobacillus panis L. reuteri, L. johnsonii, L. pontis, L. brevis

Commercial baker’s yeast +



(Gobbetti and Gaenzle, 2013. In Handbook on Sourdough Biotechnology, Springer)

1° refreshment for 15 – 20 h at ca. > 30°C

Type III sourdough. Type II liquid sourdoughs, which are dried/stabilized after preparation, are named tipe III. They are manly used at the industrial level manufactured by sourdough fermentation with subsequent water evaporation leading to dried preparations which are used as acidifier supplements and aroma carriers.

Pediococcus pentosaceus L. plantarum, L. brevis



(Gobbetti and Gaenzle, 2013. In Handbook on Sourdough Biotechnology, Springer)

1° refreshment for 15 – 20 h at ca. > 30°C

Type III

Type II

How diverse is the sourdough compared to the other starters used for making fermented foods

and beverages?


Soft and semi-soft cheeses

Streptococcus thermophilus

Commercial starters: industrial preparations composed by one or more defined microbial species

St. thermophilus Lactobacillus delbrueckii subsp. bulagaricus


Yogurt and fermented milks

Microbial biodiversity and fermented foods

Lactobacillus plantarum L. pentosus

ü  P. pentosaceus (1) ü  L. plantarum (7) ü  L. brevis (1) ü  Lc. lactis (1) ü  W. paramesenteroides

(1) ü  Leuc. mesenteroides (1)

Pane casereccio Marchigiano

ü  L. plantarum (2) ü  L. casei (1) ü  L. sanfranciscensis (1) ü  P. inopinatus (2)

ü  L. plantarum (1) ü  L. paralimentarius (4) ü  Leuc. mesenteroides (1)

Bastone di Padova

Pane Carasau

Department of Soil, Plant and Food Science, University of Bari, Italy

(Solieri et et al., 2012. Syst. Appl. Microbiol. 35, 270-277)


Inter-species biodiversity

26 Dairy farms

Natural starter: artisan preparation composed by mixed populations of different microorganisms

Whey Culture - 3%

“Bulding up the sourdough library”

•  Culture collection •  Sourdough stocks

Protocols for bread

History and main features of the bread

Back-slopping protocols

Biochemical traits of the sourdough

Lactic acid bacteria and yeasts identified and biotyped Tree Diagram for 15 Cases

Unweighted pair-group averagePercent disagreement

0% 10% 20% 30%

Linkage Distance

SC.B2SC.B13SC.B12SC.B15SC.B11SC.B10

SC.B9SC.B8

SC.B14SC.B5SC.B7SC.B6SC.B4SC.B3SC.B1

Source of sourdough biodiversity


Mapping of typical and traditional Italian baked goods


Typical and Traditional Italian sourdough breads- Library I



Typical and traditional Italian sourdough sweet baked goods - Library II

Some information on the main technology and microbiology traits


18-25°C, 24 h

At least three times Back-slopping (addition of flour and water, and

kneading)

Durum wheat flour and water

Sourdough

Durum wheat flour, salt, water Kneading (20 min)

Portioning and shaping

18-25°C, 90 min

18-25°C, 30 min

Shaping

18-25°C, 15 min

Baking (250°C, for about 1 h)

Packaging

Pane di Altamura PDO (Puglia)


Technology parameters of the 19 Italian typical/traditional breads (I)

aT.d., Triticum durum; T.a., Triticum aestivum. bFirst number indicates the length of backslopping (h), whereas the second indicates the temperature (°C) of incubation. Mature sourdough, i.e. the one obtained after the last backslopping, was subjected to analyses. cFirst number indicates the length of fermentation before baking, expressed in hours, whereas the second indicates the temperature (° C) of incubation. dFirst number indicates the length of baking step, expressed in hours, whereas the second indicates the temperature of baking. ew, wood stove; g, gas oven; e, electric oven; n.i., the type of oven is not indicated. *Baker’s yeast is used also during backslopping.

Breads Type of

floura

NaCl (%) Bakers’

yeast (%)

Sourdough

(% )

Number of

backslopping

steps

Backslopping

(time and

temperature)b

Fermentation

(time and

temperature)c

Baking

(time and

temperature)d

Ovene

Pane di Altamura

PDO

T.d. 2.0 0 20 3 10 ; 25 120 – 140; 18 –

25

60;250 w or g

Pane di Laterza T.d. 2.0 0 28 1 6 – 7 ; 25 120-360; 18 –

25

80 - 90;230 w

Pane di Matera PGI T.d. 2.5 – 3.0 0 10 2 6 – 7 ; 25 90; 18 – 25 120;280 w or g

Pane di Montecalvo

Irpino

T.a./T.d.

(20/80%)

2.0 0 20 2 6 – 7; 25 150 – 210; 18 –

25

w

Pane casereccio di

Reggio Calabria

T.a. 1.5 – 2.0 0 50 2-3 6 – 7;18 – 25 15; 220 e

Pane casereccio del

Molise

T.a. 2.0 0 30 2 – 3 10; 18 – 25 4 ;25 90;260 g

Pane casareccio di

Genzano PGI

T.a. 2.0 0.4* 1.5 1 2 ;30 100 – 120; 27 -

30

35 – 80;300 –

320

w

Bozza Pratese T.a. 0 0 20 1 15; 11 60;27 – 30 60;220 n.i.

Pane di Altopascio

Tradizionale

T.a. 0 0 40 1 24 ; 8 – 10 60 – 120;20 –

25

50 – 50; 220 –

240

n.i.

1

6

360

240

Technology parameters of the 19 Italian typical/traditional breads (II)

aT.d., Triticum durum; T.a., Triticum aestivum. bFirst number indicates the length of backslopping (h), whereas the second indicates the temperature (°C) of incubation. Mature sourdough, i.e. the one obtained after the last backslopping, was subjected to analyses. cFirst number indicates the length of fermentation (min) before baking, expressed in hours, whereas the second indicates the temperature (° C) of incubation. dFirst number indicates the length of baking step, expressed in hours, whereas the second indicates the temperature of baking. ew, wood stove; g, gas oven; e, electric oven; n.i., the type of oven is not indicated. *Baker’s yeast is used also during backslopping.

Breads Type of

floura

NaCl (%) Bakers’

yeast (%)

Sourdough

(% )

Number of

backslopping

steps

Backslopping

(time and

temperature)b

Fermentation

(time and

temperature)c

Baking

(time and

temperature)d

Ovene

Pane di Terni T.a. 0 0 14 1 8 – 16; 15 - 18 120 - 180, 27 –

30

50; 220 w

Bastone di Padova T.a. 2.0 0.6* 45 1 18 ; 25 – 30 4 ; 27 – 28 20 – 24; 200 n.i.

Coppia ferrarese PGI T.a. 2.0 0.5 10 1 12;18 – 25 70 – 90; 18 - 25 25; 210–230 n.i.

Pane casereccio

Marchigiano

T.a. 2.0 0 33 1 6 – 7;20 – 25 6;20 - 25 30 – 40; 230 n.i.

Pane di Cappelli T.d. 1.5 0 25 2 4;26 5; 28 – 30 60; 220 w or g

Moddizzosu T.d. 1.8 – 2.0 0 30 1 10 – 12; 18 – 25 3 – 4; 18 – 25 40 – 60; 250 –

280

n.i.

Pane Carasau T.d. 1.0 – 2.0 0 30 1 10 – 12;27 5;20°C 2; 540

5 – 10; 500

n.i.

Pane nero di

Castelvetrano

T.d. 1.5 0 1 1 – 2 8 – 12;

25 – 27

3 h, 18 – 25 1; 300 w

Pane di Lentini T.d. 2.0 0 20 2 10-12; 25 2 – 4 ;25 45 – 60; 250 –

300

w

Pagnotta del Dittaino

PDO

T.d. 2.0 0 15 – 18 1 5;18 – 25 4-5; 18-25 1; 230 w

1

240

360

300

180

300

180

180

300

60

60

Overview on some main technology parameters used for making typical and traditional Italian sourdough breads

Parameters % on total number of bread studied

Use of durum wheat 52

Wheat cultivar not specified 84

Use of sourdough only 73

Use of bakers’ yeast in the final resting step 26

Just one back-slopping before production 58

Addition of salt 84

Just one resting step 52

Building up the “Italian Sourdough Library” Department of Biologia e Chimica Agro-Forestale ed Ambientale – University of Bari, Italy

Biochemical characterization of the 19 Italian sourdough breads

Sourdoughs pH Lactic acid (mM)

Acetic acid (mM) Fermentation quotient

Free amino acids (mg/kg)

Pane di Altamura PDO 4.03±0.02 82.0±3.6 17.6±0.8 4.6±0.91 612.2±30.61 Pane di Laterza 4.05±0.01 79.0±3.9 15.0±0.7 5.3±0.92 669.1±33.04 Pane di Matera PGI 4.21±0.01 68.5±2.8 17.6±0.6 3.9±0.30 555.6±27.08 Pane di Montecalvo Irpino 4.17±0.03 84.0±4.2 18.8±0.9 4.5±0.93 562.2±28.11 Pane casereccio di Reggio Calabria 4.28±0.02 75.3±3.7 14.5±0.7 5.2±0.93 664.0±33.20 Pane casereccio del Molise 3.95±0.07 94.0±4.7 17.0±0.8 5.5±0.90 412.6±20.63 Pane casareccio di Genzano PGI 4.14±0.02 63.7±3.1 12.1±0.6 5.3±0.91 595.4±29.77 Bozza Pratese 3.89±0.01 90.0±4.5 18.8±0.9 4.7±0.90 570.0±28.50 Pane di Altopascio Tradizionale 3.97±0.01 87.3±2.1 6.0±0.3 14.5±0.95 590.9±29.54 Pane di Terni 3.90±0.01 75.0±3.7 16.4±0.8 4.6±0.88 613.5±30.67 Bastone di Padova 3.90±0.01 75.1±2.6 14.0±0.7 5.4±0.91 339.6±16.98 Coppia ferrarese PGI 3.89±0.01 78.5±3.9 18.3±0.9 4.3±0.91 487.1±24.33 Pane casereccio marchigiano 3.97±0.0.01 87.1±4.3 20.6±1 4.2±1.04 763.5±38.17 Pane Cappelli 3.97±0.0.02 75.0±3.7 15.0±0.7 5.0±0.90 361.7±18.08 Moddizzosu 3.87±0.11 84.2±1.9 15.6±0.5 5.4±0.91 624.2±18.48 Pane Carasau 3.97±0.08 80.5±3.1 14.4±0.4 5.6±0.91 608.7±11.50 Pane nero di Castelvetrano 3.74±0.01 78.0±3.9 17.0±0.8 4.6±0.93 1090.5±54.50 Pane di Lentini 3.89±0.02 73.4±3.6 14.5±0.7 5.1±0.89 520.2±26.00 Pagnotta del Dittaino PDO 3.70±0.02 83.2±4.1 15.2±0.7 5.5±0.90 553.4±27.65


Most (68%) of the sourdoughs have values of pH lower than 4.0

Almost all (except one) the sourdoughs have a quotient of fermentation of 5.5 or below Most (79%) of the sourdoughs have concentration of free amino acids higher than 500 mg/kg

Overview on some biochemical parameters of the typical and traditional Italian sourdough breads


DGGE profiles of 19 Italian sourdough breads: primers LS2/NL1 for yeasts group

A B C D E F G H I L M N O P Q R S T UMk Mk Mk

1

34

5

6

2

S SSS SS

1.  Triticum sp.

2. Candida humilis/Kazachstania barnettii/Kazachstania exigua

3. Candida humilis/Kazachstania barnettii 4. Candida humilis/Kazachstania barnettii 5. Saccharomyces cerevisiae 6. Saccharomyces cerevisiae

Yeast cell numbers in Italian sourdough breads

log

cfu/

g

3

4

5

6

7

8

9

log

cfu/

g

3

4

5

6

7

8

9

MEASDA

log

cfu/

g

3

4

5

6

7

8

9

log

cfu/

g

3

4

5

6

7

8

9

Building up the “Italian Sourdough Library” Department of Biologia e Chimica Agro-Forestale ed Ambientale – University of Bari, Italy

Identification of lactic acid bacteria by 16S rDNA gene and RAPD-PCR analyses Sourdoughs Isolates Strains Species (number of strains)

Pane di Altamura PDO 39 14 W. cibaria (12); W. confusa (1) Pane di Laterza 17 9 L. sanfranciscensis (4); Leuc. citreum (5) Pane di Matera PGI 18 5 L. plantarum (1); Leuc. citreum (4) Pane di Montecalvo Irpino 19 8 L. sanfranciscensis (7) Pane casereccio di Reggio Calabria 19 11 L. sanfranciscensis (1); L. sakei (3) Leuc.

mesenteroides (5) Pane casereccio del Molise 24 8 L. sanfranciscensis (7) Pane casareccio di Genzano PGI 22 12 L. plantarum (5); P. pentosaceus (5) Bozza Pratese 28 6 L. sanfranciscensis (4); L. paralimentarius (1) Pane di Altopascio Tradizionale 25 8 L. gallinarum (7); L. sanfranciscensis (1) Pane di Terni 20 9 L. sanfranciscensis (1); L. plantarum (3); Lc. Lactis (3);

L. brevis (1) Bastone di Padova 25 8 L. plantarum (1); L. paralimentarius (4); Leuc.

mesenteroides (1) Coppia ferrarese PGI 28 7 L. plantarum(4); L. paralimentarius (2) Pane casereccio marchigiano 14 7 L. plantarum (2); L. casei (1); L. sanfranciscensis(1); P.

inopinatus (2) Pane Cappelli 18 10 L. sanfranciscensis (6); L. plantarum (2) Moddizzosu 25 17 L. sanfranciscensis(3); L. plantarum(1); L. brevis(7); L.

rossiae (3); P. argentinicus (1) Pane Carasau 29 13 P. pentosaceus(1); L. plantarum(7); L. brevis;(1) Lc.

lactis (1); W. paramesenteroides (1); Leuc. mesenteroides (1)

Pane nero di Castelvetrano 32 9 L. spicheri (3); L. paralimentarius (4); Pane di Lentini 20 9 L. sanfranciscensis (1); L. paralimentarius (4); L.

namurensis (1) Pagnotta del Dittaino PDO 10 7 L. sanfranciscensis(4); E. durans(3)

Lactic acid bacteria cell numbers in typical and traditional Italian sourdough breads

log

cfu/

g

5

6

7

8

9

10

log

cfu/

g

5

6

7

8

9

10

mMRSSDBMRS 5M17


The diversity of lactic acid bacteria (LAB) was very elevated (twenty different species were identified)

None of the sourdoughs had the same population of lactic acid bacteria The number of lactic acid bacteria species varied from one to six per sourdough

The cell number of lactic acid bacteria markedly varied between sourdoughs (107 to 109/g)

Overview on some microbiological (LAB) parameters of the typical and traditional Italian sourdough breads



Pane di Cappelli

Pane Carasau

Pane di Lentini

-12 -10 -8 -6 -4 -2 0 2 4 6 8 10

Factor 1: 52,86%

-6

-5

-4

-3

-2

-1

0

1

2

3

4

5

6

7

Factor 2: 12,22%

Pane nero di Castelvetrano Moddizzosu

Pane di Terni


Pagnotta del Dittaino PDO

Pane casereccio del MolisePane di Montecalvo Irpino

Pane casereccio di Reggio CalabriaPane di Altamura PDO Pane casereccio di

Genzano PGI

Bozza PratesePane di Laterza

Pane di Matera PGI Bastone di Padova

Coppia ferrarese PGI

Pane di Altopascio tradizionale

Pane di Cappelli

Pane Carasau

Pane di Lentini

-12 -10 -8 -6 -4 -2 0 2 4 6 8 10

Factor 1: 52,86%

-6

-5

-4

-3

-2

-1

0

1

2

3

4

5

6

7

Factor 2: 12,22%

Pane nero di Castelvetrano Moddizzosu

Pane di Terni


Pagnotta del Dittaino PDO

Pane casereccio del MolisePane di Montecalvo Irpino

Pane casereccio di Reggio CalabriaPane di Altamura PDO Pane casereccio di

Genzano PGI

Bozza PratesePane di Laterza

Pane di Matera PGI Bastone di Padova

Coppia ferrarese PGI

Pane di Altopascio tradizionale

PCA based on the composition of flour and microbial community


Buondì

Panettone P

Cornetto

Panaredda

Pandoro

Nadalin

Resta di ComoPanettone basso

-7 -6 -5 -4 -3 -2 -1 0 1 2 3 4 5 6

Factor 1: 27,19%

-6

-5

-4

-3

-2

-1

0

1

2

3

4

5

Factor 2: 16,77%

Buccellato di LuccaMbriagotto

Torcolo SCPizza di Pasqua

Panettone LBiscotti di Lagaccio

Pandolce GenoveseColombaVeneziana

Ciambella di Mosto

PCA based on the process parameters, biochemical characteristics and microbial community

The diversity of lactic acid bacteria (LAB) was more limited (eleven different species were identified, but eight species were found just occasionally).

Lactobacillus sanfranciscensis was identified in all the sourdoughs, except for Panaredda and Torcolo di San Costanzo. For eleven sourdoughs, all the LAB isolates were allotted to L. sanfranciscensis.

The cell number of lactic acid bacteria markedly varied between sourdoughs (106 to 109/g)

Overview on some microbiological (LAB) parameters of the traditional Italian sourdough sweet leavened baked goods


The main challenge in sourdough biotechnology is to get the right consortium of lactic acid bacteria

Lactic acid bacteria mainly determine the superior quality of the sourdough baked goods



Lactobacillus sanfranciscensis (species)

(strains belonging to same species)

(s13) (s16)

(s14)

(s18)

(s17)

(s24)

(s23)

(s22) (s21)

(s15) (s20)

(s19)

Lactobacillus sanfranciscensis




Nowadays...cultural heritage


- The collections of the Library of Congress include more than 32 million catalogued books and other print materials in 470 languages -

Library of Congress (USA): the largest library in the world ...

A l i b r a r y i s t h e conservatorship of the wise souls, where not only you breathe the dust of centuries, but where you feel the thinking of men ...


- The collections of the sourdough library include, at this time, about 40 Italian sourdoughs and about 400 strains of lactic acid bacteria and yeasts -

Library (Saint-Vith): the first library in the world of sourdoughs ...

A sourdough library is the conservatorship of al ive ingredients, where not only you breathe the cultural heritage of centuries, but where you can still feel the odour of the ferment of life ...

inauguration sourdough library · 2019-02-18 · kneading (20 min) portioning and shaping 18-25°c,...

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