Malt Adjunct Hot Wort Tank

(rice, corn, wheat)

Mill
Plate Cooler
Cereal Cooker

Mash Mixer Yeast + 02

Syrup Hops Fermenter

Lauter Tun or
Mash Filter
Kettle Centrifuge
Spent Grain
Aging
Packaging Bright Beer
(bottle, can, keg) Tank Carbonation Filtration

Fig. 7.1. The brewing process.

53
Fig. 7.2. Principal brewing raw materials.

54
 Barley

Steeping
Grain hydration

Germination
Enzyme generation

Kilning
Drying and curing

Malt
Fig. 7.6 Typical barley malting process.

55
 Aleurone Scutellum
Endosperm Embryo Husk

Fig. 7.4. The barley grain.

56
Fig. 7.5. Structure of the barley aleurone layer.

57
 80
To lauter tun
or mash filter

Saccharification at 65°C
Temperature °C

60

Protein and
Glucan conversion
at 52-55°C

40
0 10 20 30 40 50 60 70
Time, minutes

Fig. 7.6. Temperature profile of a typical infusion mashing programme.

58
Fig. 7.7. The enzymatic hydrolysis of starch.

59
Fig. 7.8. Structure of maltose and maltotriose.

60
 90

80 Glucose
Carbohydrate (g/L)

Fructose
70 Maltose
60 Maltotriose
Dextrins
50

40

30

20

10

0
0 24 48 72 96 120 144

Fermentation time (hours)

Fig. 7.9. Order of uptake of wort sugars by the yeast cell.

61
Fig. 7.10. Uptake and metabolism of maltose and maltotriose by the yeast cell.

62
Fig. 7.11. Effect of osmotic pressure [25% (w/v) sortibol*] on the uptake of glucose, maltose
and maltotriose by a lager strain and an ale strain .

63
Fig. 7.12. Effect of ethanol [10% (v/v) ethanol*] on the uptake of glucose, maltose and
maltotriose by a lager strain and an ale strain .

64
 200
180
160
Sug Sugar concentration (g/L)

140
120
100
80
60
40
20
0

12°Plato (30% 20°Plato (30% 20°Plato (30% VHM)

maltose syrup) maltose syrup) syrup
Glucose and fructose Maltose and maltotriose

Fig. 7.13. Wort sugar profile.

65
Fig. 7.14. The Heriot-Watt University, Edinburgh, Scotland brewing pilot plant.

66
 40
Ethyl acetate concentration

30
(mg/L)

20

10

0
0 40 80 120 160 200 240

Fermentation Time (hours)

12ºPlato (30% MS) 20ºPlato (30% MS) 20ºPlato (30% VHMS)

Fig. 7.15. Ethyl acetate concentration in fermenting worts of differing gravities and sugar
composition.

67
Fig. 7.16. Isoamylacetate concentration in fermenting worts of differing gravities and sugar
composition.

68
 S. bayanus

low 72%

S. paradoxus low S. pastorianus

w

50% 50%

S. cerevisiae

Fig. 7.17. The Saccharomyces sensu structo group for ale and lager strains.

69
 A B
3 9
Ale strains 3
Maltotriose (g/100 ml)

Maltose (g/100 ml)

Ale strains 5 8

2.5 Ale strains 11 7

Ale strains 30 6
Lager strains 15
5
2 Lager strains 22
4
Lager strains 27
Lager strains 42 3

1.5 2

1 0
0 24 48 72 96
0 2 4 6 8 10 12 16 20 23 26 29 31

Fermentation time (hours) Fermentation time (hours)

Fig. 7.18. Maltotriose (A) and maltose (B) uptake profiles from a 16°Plato wort.

70
 20
M AL2/M AL2;
16 M AL4/M AL4
Degree Plato

M AL2/mal2;
12 mal4/mal4

0
0 24 48 72 96
Fermentation time (hours)
*Shaking fermentations at 21°C

Fig. 7.19. Fermentation profile of a 16°Plato wort with a diploid yeast strain containing
multiple maltose (MAL) genes.

71
 100
M AL2/M AL2;
M AL4/M AL4
75
M AL2/mal2;
Maltose (g/L)

mal4/mal4
50

25

0
0 24 48 72 96

Fermentation time (hours)

*Shaking fermentations at 21°C

Fig. 7.20. Uptake of maltose from 16°Plato wort by a diploid yeast strain containing
multiple (MAL) genes.

72
 14

12 AMYLOLYTIC
Degrees Plato

YEAST
10 CONTROL YEAST

0
0 30 70 120 165 210 240 290
Fermentation time (hours)

Fig. 7.21. Effect of glucoamylase in a brewing strain during wort fermentation.

73
 14

12 AMYLOLYTIC
Degrees Plato

YEAST
10 CONTROL YEAST

0
0 30 70 120 165 210 240 290
Fermentation time (hours)

Fig. 7.22. Production of glucose in pasteurized beer during storage at 21°C.

74
Fig. 7.23. Brewing Research Foundation International – Nutfield Lyte.

75
 Amino Acid Absorption Pattern
during Wort Fermentation
3.5 4.5
3.0 4.0
3.5 3rd
1st 2.5 3.0 Group,
and 2.0 2.5 Proline
2nd 1.5 2.0 (mmol/l)
groups 1.5
1.0
(mmol/l) 1.0
0.5 0.5
0.0 0.0
0 19 24 43 48 67 72 96

Fermentation Time (hours)

1st group 2nd group 3rd group Proline

Fig. 7.24. Amino acid absorption pattern during wort fermentation.

76
Fig. 7.25. Intracellular concentration of glycogen and lipids in a lager yeast strain during
fermentation of a 15°Plato wort.

77
 Carbohydrate
Ethanol
Acetic Acid
Pyruvate

Ethanol Acetic Acid

Enzymatic
Conversion
α-Acetohydroxy-
α-Acetolactate butyrate Passive
Diffusion
Plasma
Membrane Valine Iso- Non-Enzymatic
leucine leucine Decomposition

α-Acetohydroxy-
α-Acetolactate butyrate

Diacetyl Pentanedione

Fig. 7.26. Fermentation of diacetyl and 2.3-pentandeione as by-products of pathways leading

to the formation of valine and isoleucine.

78
 Diacetyl
Plasma
Diacetyl Membrane

Enzymatic
Conversion
Acetoin
Plasma
Diffusion
Butanediol

Acetoin
Butanediol

Fig. 7.27. Reduction of diacetyl to acetoin and 2,3-butanediol.

79
 ALDC Yeast
0.8 Control Yeast
0.7
D iacetyl( m g/L)

0.6
0.5
0.4
0.3
0.2
0.1
0
0 50 100 150 200
Fermentation time (hours)

Fig. 7.28. Effect of alpha acetolactate decarboxylase (ALDC) expression in a brewing yeast
strain on diacetyl metabolism during wort fermentation.

80
 ALDC Yeast Control Yeast

16
14
12
Degrees plato

10
8
6
4
2
0
0 50 100 150 200
Fermentation time (hours)

Fig. 7.29. Effect of alpha acetolactate decarboxylase (ALDC) expression in a brewing yeast
strain on overall fermentation rate during wort fermentation.

81
 LIST OF TABLES

Table 7.1 Typical barley malt analysis. (Briggs, 1998b).

Table 7..2 Typical sugar spectrum of wort. (Stewart, 2006).

Table 7.3 Influence of wort gravity on beer ester levels. (Younis and Stewart, 1999).

Table 7.4 Ethyl acetate and isoamyl acetate produced by brewing yeast strains during

fermentation of synthetic media. (Younis and Stewart, 1999).

Table 7.5 Sugar composition of brewing syrups. (Priest and Stewart, 2006).

Table 7.6 Comparison of ale versus lager consumption (%) trends in Ontario, Canada

(1970 – 2007). (Steiner, 2009).

Table 7.7 Comparison of ale versus lager consumption (%) trends in the United Kingdom

(1970 – 2007). (Steiner, 2009).

Table 7.8 Order of wort amino acids and ammonia uptake during fermentation. (Jones and

Pierce, 1964).

82
 LIST OF FIGURES

Fig. 7.1. The brewing process.

Fig. 7.2. Principal brewing raw materials.

Fig. 7.3. Typical barley malting process.

Fig. 7.4. The barley grain. (Briggs, 1998b).

Fig. 7.5. Structure of the barley aleurone layer. (Briggs, 1986).

Fig. 7.6. Temperature profile of a typical infusion mashing programme. (Briggs, 1986).

Fig. 7.7. The enzymatic hydrolysis of starch.

Fig. 7.8. Structure of maltose and maltotriose.

Fig. 7.9. Order of uptake of wort sugars by the yeast cell. (Stewart, 2006).

Fig. 7.10. Uptake and metabolism of maltose and maltotriose by the yeast cell. (Zheng et al,

1994).

Fig. 7.11. Effect of osmotic pressure [25% (w/v) sortibol*] on the uptake of glucose, maltose

and maltotriose by a lager strain and an ale strain . (Pratt et al, 2003).

Fig. 7.12. Effect of ethanol [10% (v/v) ethanol*] on the uptake of glucose, maltose and

maltotriose by a lager strain and an ale strain . (Pratt et al, 2003).

Fig. 7.13. Wort sugar profile. (Stewart, 2006).

Fig. 7.14. The Heriot-Watt University, Edinburgh, Scotland brewing pilot plant.

Fig. 7.15. Ethyl acetate concentration in fermenting worts of differing gravities and sugar

composition. (Younis and Stewart, 1999).

Fig. 7.16. Isoamylacetate concentration in fermenting worts of differing gravities and sugar

composition. (Younis and Stewart, 1999).

83
Fig. 7.17. The Saccharomyces sensu structo group for ale and lager strains. (Pederson, 1995).

Fig. 7.18. Maltotriose (A) and maltose (B) uptake profiles from a 16°Plato wort. (Zheng et al,

1994).

Fig. 7.19. Fermentation profile of a 16°Plato wort with a diploid yeast strain containing

multiple maltose (MAL) genes. (Stewart et al, 1995).

Fig. 7.20. Uptake of maltose from 16°Plato wort by a diploid yeast strain containing multiple

(MAL) genes. (Stewart et al, 1995).

Fig. 7.21. Effect of glucoamylase in a brewing strain during wort fermentation. (Erratt and

Stewart, 1981).

Fig. 7’.22. Production of glucose in pasteurized beer during storage at 21°C. (Erratt and

Stewart, 1981).

Fig. 7.23. Brewing Research Foundation International – Nutfield Lyte. (Baxter, 1995).

Fig. 7.24. Amino acid absorption pattern during wort fermentation. (Lekkas et al, 2005).

Fig. 7.25. Intracellular concentration of glycogen and lipids in a lager yeast strain during

fermentation of a 15°Plato wort. (Quain et al, 1981).

Fig. 7.26. Fermentation of diacetyl and 2.3-pentandeione as by-products of pathways leading

to the formation of valine and isoleucine. (Wainwright, 1973).

Fig. 7.27. Reduction of diacetyl to acetoin and 2,3-butanediol. (Wainwright, 1973).

Fig. 7.28. Effect of alpha acetolactate decarboxylase (ALDC) expression in a brewing yeast

strain on diacetyl metabolism during wort fermentation. (Yamano et al, 1994).

Fig. 7.29. Effect of alpha acetolactate decarboxylase (ALDC) expression in a brewing yeast

strain on overall fermentation rate during wort fermentation. (Yamano et al, 1994).

84