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09603085/03/$23.50+0.00 # Institution of Chemical Engineers Trans IChemE, Vol 81, Part C, March 2003

THE USE OF LABORATORY-SCALE FERMENTATIONS AS A TOOL FOR MODELLING BEER FERMENTATIONS

N. HEPWORTH1 , A. K. BROWN2 , J. R. M. HAMMOND 2 , J. W. R. BOYD1 and J. VARLEY1
1

Department of Chemical Engineering and Chemical Technology, Imperial College London, London, UK 2 Brewing Research International, Nut eld, Surrey, UK

here have so far been relatively few attempts reported in the literature to develop models for prediction of beer fermentation at large scale as a function of key process conditions. Important outputs would be concentrations of key avour components, which are important determinants of product quality. Such models would be useful for their predictive power, and in the longer term for process optimization. In this paper, an approach for linking the effect of mixing, based on power input, to kinetic models based on small-scale homogenous fermentations is presented. The kinetic models themselves are useful as they can be used for prediction of ethanol, substrate, ester and higher alcohol yield coef cients, as a function of temperature, pitching rate, gravity and initial dissolved oxygen concentration in the wort, based on on-line measurements of CO2. On-line measurement of CO2 is relatively easy and inexpensive and so models of this kind should provide a useful rst stage towards developing more rigorous models for the brewing industry. Keywords: beer; kinetics; effectiveness; hydrodynamics; mathematical modelling.

INTRODUCTION Considering the established nature of the brewing industry, there is surprisingly little published information on the modelling and control of industrial-scale beer fermentations. Models which could be used to predict, over a range of scales of operation, the development of yeast growth, ethanol and avour production, particularly as a function of key process parameters, should be of great value to the brewing industry. Such models would, together with appropriate control strategies, allow for more consistent production and could be used for process optimization. The utility of modelling approaches for beer fermentation proposed to date has been restricted because: most models have been based on data from bench scale fermenters, in which there are few mass transfer and mixing limitations, unlike processes at industrial scale (400 m3); those models which have been developed have been validated for only a narrow range of process conditions; data used for model development have primarily been limited and taken off-line; models which allow rigorous optimization would be extremely complex, as full account would have to be taken of kinetics (including effect of key process variables) and of the three-phase hydrodynamics, which are particularly complex because the ow is driven by timevarying CO2 evolved during the process. 50

The key elements of a successful model will be: (1) (2) (3) (4) (5) accurate prediction of cell growth, substrate uptake and ethanol production; accurate prediction of the main beer avour compounds; robustness of model predictions to changes in the key process conditions (e.g. temperature, pitching rate, pH) dependence on input parameters, which can be measured relatively easily and on-line; applicability of models over a range of fermenter scales, from laboratory to industrial scale.

In this paper a systematic approach towards development of a rather simplistic model, which could allow prediction of key characteristics of beer fermentations, is described. Importantly, those in the brewing industry could use this type of approach readily without the need for complex mathematical modelling of ow patterns in large-scale fermenters. This approach involves: (1) A kinetic model based on laboratory experimental data for impeller agitated fermentations, which are assumed to be well mixed. These models build on those published previously for fermentations driven by CO2 evolution only (no mention is made by these authors of any additional agitation during fermentation; Trelea et al., 2001; Titica et al., 2000). Linkage of this kinetic model to larger-scale processes through the development of a relationship between

(2)

LABORATORY-SCALE FERMENTATIONS (i) the reaction rate in the well-mixed homogenous bioreactor with no diffusional limits and (ii) reaction rates in larger-scale processes. Alternative approaches have been proposed, for example, a scale-down approach has been described as part of a methodology for optimizing fermenter dynamics and design, which included the following steps: (1) regime analysis of the (proposed) process on a product scale; (2) simulation of the rate-limiting mechanisms on a laboratory scale; (3) optimization and modelling of the process on a laboratory scale; and (4) implementation of the process on a production scale by transformation of the optimised laboratory conditions (Luckiewicz, 1978). Other models, for example, for predicting CO2 generation and CFD models applicable to beer fermentations, have also been described, with the focus on modelling hydrodynamics rather than integrating kinetic and hydrodynamic models (Luckiewicz, 1978; Luyben, 1997). The development of kinetic models for well-mixed reactors with no diffusional limits will rst be described, followed by consideration of methods for utilising these models for larger-scale processes. Monitoring of beer fermentations typically includes off-line measurement of wort density, ethanol concentration and a selection of other fermentation products including key avour compounds. The maintenance of a particular avour pro le is obviously very important in beer production. Typically, pH, temperature, dissolved oxygen and pressure are the only variables measured on-line. It is generally accepted that the kinetic mechanisms underlying ethanol and avour production in beer fermentations are relatively well understood. Yeast growth is generally described in terms of Monod kinetics, i.e. dX mX dt (1)

51

conditions at laboratory scale (Trelea et al., 2001; Titica et al., 2000). The operating conditions they considered were: fermentation temperatures of 13, 10 and 16 C; pressures of 800, 450 and 50 mbar; and initial yeast concentrations of 20 106, 10 106 and 5 106 million cells per ml. The following relationships were developed for ethanol, substrate, higher alcohol and ester concentration as a function of time: Ethanol E(t) YE=C Cp (t ) Substrate S (t ) S0 YS =C Cp (t ) Higher alcohols Alc(t ) YAlc=C Cp (t ) Esters If Cp (t ) Cdc,m (phase 1) Est (t ) YEst=C ,1 Cp (t) Est (t ) YEst=C ,1 Cdc,m If Cp (t ) Cdc,m (phase 2) YEst=C ,2 [Cp (t) Cdc,m ] (6) (5) (4) (3) (2)

where X is the yeast concentration, t is time and m is a speci c growth rate constant. The form of this equation may be modi ed to take account of, for example, product (ethanol) inhibition, substrate inhibition and self-inhibition (for a review of kinetics of wine fermentation see Mar n, 1999). The production of ethanol and various avour compounds including esters and fusel alcohols has been investigated; the production of these avour compounds is linked to yeast growth (Garc a et al., 1994a). Recently, kinetic models have been written in terms of evolution of CO2; this is particularly valuable because CO2 evolution can be measured relatively easily on-line. Linear relationships between the production of higher alcohols and CO2 have been reported (Titica et al., 2000). A two-stage relationship between the production of esters and CO2 was found, with a slow rst phase of synthesis, followed by a second accelerated phase. The concentrations were expressed through the use of different yield coef cients for the two phases. The transition from slow to accelerated phase corresponded to the time at which the rate of CO2 production was a maximum, regardless of the operating conditions. This work was extended to include a model for the fermentation pro le including ethanol production and substrate consumption (Trelea et al., 2001). Relationships between CO2 production and ethanol production and substrate consumption have been developed and yield coef cients have been modelled for a range of operating Trans IChemE, Vol 81, Part C, March 2003

where E(t ), S (t ), Alc(t ) and Est(t) are ethanol, substrate, higher alcohol and ester concentrations, respectively, Cdc,m represents the concentration of CO2 produced corresponding to the maximum CO2 production rate, Cp(t) represents the total amount of produced CO2; S0 is the initial substrate concentration and YE= C, YS= C, YAlc=C, YEst=C,1 and YEst=C,2 are the yield coef cients for ethanol, substrate, higher alcohol, ester (phase 1) and ester (phase 2), respectively. YAlc= C was considered to vary with temperature, the dissolved fraction of CO2 and X0 and YEst= C,1 and YEst=C,2 to vary with temperature, pressure and X0 2 . In addition to the individual component relationships described above, the relationships for ethanol and substrate were combined to give the following equation for the rate of production of CO2 as a function of time: S [Cp (t)] dCp (t ) 1 umax KS S [Cp (t )] 1 KI E[Cp (t)]2 dt [Cp (t ) C0 X0 ]

(7)

where umax is the maximum speci c CO2 production rate, KS is the substrate limitation coef cient, KI is the product inhibition coef cient, C0 is the initial production rate coef cient and X0 is the initial yeast concentration, umax being a function of temperature, pressure and X0. The effect of liquid mixing and fermenter scale has been considered previously (Garc a et al., 1993, 1994b, 1995). The degree of mixing (or homogeneity) throughout the fermentation process has been correlated in terms of CO2 production rate (Garc a et al., 1994b). The work was extended by introduction of the concept of an effectiveness factor Z, which was de ned as the ratio of the rate of reaction in reactor under consideration to the rate of the reaction in a homogeneous reactor (i.e. a reactor with no diffusional limits; D az et al., 1996).

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HEPWORTH et al.
Table 1. Run numbers and conditions for all 1 l fermentation experiments. Run number 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 Temperature ( C) 12 21 21 28 28 21 21 21 28 12 12 21 21 21 12 21 21 28 12 21 12 21 28 21 21 28 28 28 21 21 28 28 28 12 Pitching rate (g L1) 0.80 0.36 0.30 0.30 0.35 0.32 0.40 0.14 0.36 0.20 1.04 0.42 0.11 0.50 0.07 0.84 0.58 0.21 0.18 0.61 1.22 0.88 0.88 2.11 1.92 0.89 1.06 0.21 1.00 1.18 0.21 0.37 2.18 2.03 Gravity ( Plato) 13 19.25 15.25 14.75 19.5 14.5 10.75 14.5 13.75 14.75 10 14.75 18.75 19 15.25 14.5 14.75 10.75 15 14.5 17.25 9.75 13.75 9.75 17.25 18.5 14.75 10.75 18.75 15.25 15 18.75 17.75 14.75 Initial DO level in the wort (% air saturation) 50 0 50 100 50 0 50 50 50 100 50 100 50 50 50 50 100 100 0 0 50 50 50 0 100 0 50 50 0 50 100 50 50 50

In this paper, research is described in which, the kinetic models discussed above have been applied to data collected from 1 l fermentations at Brewing Research International (BRi, Surrey, UK) including on-line CO2 data to predict substrate uptake and ethanol, higher alcohol and ester production for a range of temperatures (1228 C), pitching rates (0.112.18 g l1), gravities (1319.25 Plato), and initial dissolved oxygen concentration in the wort (0100% air saturation). A second part of the model is to link the kinetic data obtained on small-scale experiments to industrial large-scale fermentations. This will be through the use of an effectiveness factor based on CO2 emissions, which due to its relative simplicity has the potential to be used widely by those in the brewing industry. METHODS AND MATERIALS Yeast Strain The lager strain NCYC 1324, was used in this study. Permanent stocks are kept in the BRi Yeast Culture Collection at 196 C in liquid nitrogen. Working cultures are maintained on MYGP agar slopes at 4 C. Yeast Propagation All wort (standard BRi lager wort, 16 Plato) was produced in the BRi pilot brewery. Yeast propagation was initiated by inoculating 10 ml of fresh lager wort with a sterile loop of yeast from an agar slope. This culture was shaken at 150 rpm at 25 C for 24 h. Subsequently, the culture was transferred into a ask containing 100 ml lager wort and shaken (150 rpm) at 25 C for 24 h. Yeast cells were then enumerated using a haemocytometer and a methylene blue stain using the standard Institute of Brewing (IOB) method. The desired pitching rate for the fermentation was obtained by diluting the inoculum with fresh lager wort. An inoculum volume of 100 ml was used for all fermentations. Batch Yeast Fermentation Two litre Applikon fermenter systems (sterilized prior to use at 121 C for 15 min) were used in this study. All process data including pH (Applikon, UK), dissolved oxygen (Broadley James, USA), carbon dioxide evolution (Sierra Instruments, USA) and temperature were logged on-line using BioXpert software (Applikon, UK) with an Applikon ADI 1035 BioConsole controlled by an Applikon ADI 1030 BioController. Temperature was controlled throughout the fermentations via continuous circulation of water through the fermenter outer jacket, employing a B. Braun (Reading, Berks, UK) heater=chiller unit. Dissolved oxygen and pH were not controlled at any set-point values. A 900 ml sample of fresh sterile wort was pumped into the fermenter and 100 ml of inoculum (diluted to the required yeast concentration) added at the onset of the fermentation. The fermentations were stopped once the speci c gravity had reached 2.5 Plato. Thirty-four fermentations were run for a range of four process variables (temperature, pitching rate, gravity and initial dissolved oxygen concentration in the wort); run numbers and process conditions used are shown in Table 1.

Speci c Gravity and Substrate Measurement Speci c gravity was measured using a Kyoto Instruments DA-300 Speci c Gravity Meter. Cell counts and viability measurements were performed as described in the IOB Methods of Analysis. All speci c gravity and cell measurements were taken during the course of the fermentation; speci c gravity measurements were converted to density and then to Plato according to:

Plato

(1000 density) 999448 4:08745

The sugar (substrate) concentration was taken as the difference between the measured density and the density at the end of the fermentation (2.5 Plato). Chemical Analysis Samples were taken at the end of the fermentation (once speci c gravity reached 2.5 Plato) and analysed for avour volatiles and ethanol. Vicinal diketones were measured by GLC as described in the European Brewery Convention Recommended Methods of Analysis using a 50 m 0.22 mm internal diameter SGE BP20 0.25 (WCOT) fused silica column. Higher alcohols and esters were measured by GLC as described in the IOB Methods of Analysis using a 60 m 0.25 mm internal diameter CP Wax S7-CB (WCOT) fused silica column. These analyses were Trans IChemE, Vol 81, Part C, March 2003

LABORATORY-SCALE FERMENTATIONS performed using a Perkin Elmer Autosystem XL Gas Chromatograph tted with a HS40 Headspace Analyser and a FID. Ethanol was measured essentially as described in the IOB Methods of Analysis on a 15 m 0.53 mm internal diameter HP-Wax cross-linked polyethylene glycol column using a Hewlett Packard 5890A Gas Chromatograph tted with a HP6890 Series Injector and an FID. Pilot-s cale Fermentations The only larger scale data available and which included on-line CO2 measurements was for 11 litre cylindrico-conical fermentations run at BRi. The process conditions for the fermentation considered here were initial dissolved oxygen concentration in the wort 100% air saturation; pitching rate 2.5 g l1, gravity 9.25 Plato and temperature 20 C. RESULTS AND ANALYSIS The fermentation runs with values for the four process variables (temperature, pitching rate, gravity and initial dissolved oxygen concentration in the wort) under consideration here are shown in Table 1. For comparison of the data runs, the pitching rate, gravity, temperature and dissolved oxygen level (DO) were normalised according to the following equations: 1b In with c I Imin c max 2 I Imax b min 2 and (8)

53

time point residuals were summed and averaged and the values of umax, KI and KS were found which minimized this total residual (using a NewtonRaphson optimization technique within the package Excel). It was assumed that KI and KS do not vary with operating conditions as has been reported previously (Trelea et al., 2001); the values taken were those calculated for run 31 (this being the run closest to central values of all variables); values for this run were found to be KI 0.2431 and KS 17.0886. In all analysis here it was assumed that C0 0. Also, as described previously (Titica et al., 2000), account was taken of CO2 that would be produced initially but would not be evolved until saturation is reached, as it would initially be dissolved in the wort. For runs 230 and 3235, the yield coef cients for the avour components under consideration and umax were determined as linear functions of temperature (T), pitching rate (P), substrate concentration (S) and initial dissolved oxygen concentration in the wort (D) and pairwise interactions as described in equation (9): X X Y a0 aI In aT ,I Tn , In
I S ,D

where In represents the normalized value of the factor (I varies between Imin and Imax). The normalized values therefore ranged from 1 to 1. For each run, the yield coef cients for ethanol, substrate and avour compounds were identi ed from experimental data by regression analysis, using equations (2)(6). (The experimental data for CO2 evolution was converted to a cumulative CO2 pro le. From this pro le, values of Cp for each sampling time point were available; from these values and concentration data, yield coef cients could then be calculated for each run). For each measurement of CO2 evolution (every 10 min), the residual between the experimental evolution rate (dCp=dt) and the predicted evolution rate [using equation (7)] was calculated [i.e. (experimental predicted)2]; these

where subscript n denotes normalized value of factor [normalized according to equation (8)] and a values are the coef cients with respect to variables given in the subscript. Where the subscript for a is a product of two variables, this indicates the coef cient is for an interaction between these two process variables (higher than pairwise interactions were found to be insigni cant). The calculated coef cients for umax and avour compounds: ethyl acetate, iso butyl acetate, iso amyl acetate, ethyl hexanoate, n-propanol, iso-butanol and iso-amyl alcohol are shown in Tables 2 and 3. Factor normalization is valuable as it allows determination of relative importance of factors on yield coef cients. The larger the coef cient, the more in uential the factor. The value of the sign for the coef cient indicates either an increase in the coef cient with an increase (positive sign) or decrease (negative sign). For run 31 values, yield coef cients and umax calculated using equations (2)(7) (as described above) were compared with a second set of values for yield coef cients and umax calculated using equation (9) and the values of coef cients given in Tables 2 and 3. These two sets of values of yield

I T ,P,S ,D

I P,S ,D

aP,I Tn , In aSD Sn Dn

(9)

Table 2. Yield coef cient parameters for fermentation pro le and higher alcohols. CO2 pro le vmax (h1) a0 aT aP aS aD aTP aTS aTD aPS aPD aSD 0.4814 0.1088 0.0964 0.1391 0.1581 0.0646 0.2280 0.1559 0.1653 0.1096 0.0445 Ethanol Yeth=C [l(100 l1)] 0.5914 0.3770 0.2529 0.0007 0.0080 0.3592 0.0514 0.0434 0.1122 0.1473 0.0450 Substrate Ysubs= C (g l1) 5.3363 4.2316 3.6863 0.8572 0.7585 4.9073 0.5159 0.8992 1.8500 2.2612 0.1439 n-Propanol Yalc=C (mg l1) 2.599 1.523 0.497 0.612 0.263 0.949 0.396 0.245 0.329 0.553 0.496 Iso-butanol Yalc=C (mg l1) 2.814 1.546 0.548 0.948 0.891 0.973 0.396 1.313 0.039 0.808 1.004 Iso-amyl alcohol Yalc= C (mg l1) 11.845 7.035 4.463 4.090 1.469 7.084 4.429 1.959 0.321 0.754 3.111

Units for coef cient parameters can be determined from equation (9).

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HEPWORTH et al.
Table 3. Yield coef cient parameters for esters. Ethyl acetate YEst= C,1 (mg l1) a0 aT aP aS aD aTP aTS aTD aPS aPD aSD 0.961 0.401 0.258 0.106 0.467 0.458 0.128 0.737 0.232 0.298 0.046 YEst= C,2 (mg l1) 4.189 4.349 2.130 3.859 0.100 2.161 7.401 3.088 0.219 1.300 3.238 Iso-butyl acetate YEst=C,1 (mg l1) 0.002 0.002 0.006 0.003 0.001 0.001 0.001 0.006 0.005 0.004 0.007 YEst=C,2 (mg l1) 0.013 0.014 0.015 0.007 0.019 0.024 0.019 0.059 0.003 0.018 0.003 Iso-amyl acetate YEst=C,1 (mg l1) 0.056 0.008 0.034 0.009 0.002 0.024 0.008 0.022 0.069 0.053 0.087 YEst= C,2 (mg l1) 0.367 0.263 0.187 0.237 0.013 0.224 0.417 0.248 0.076 0.086 0.178 Ethyl hexanoate YEst= C,1 (mg l1) 0.011 0.003 0.001 0.006 0.001 0.002 0.005 0.012 0.005 0.004 0.001 YEst= C,2 (mg l1) 0.033 0.026 0.017 0.016 0.003 0.036 0.023 0.027 0.001 0.000 0.012

Table 4. Yield coef cient parameters for the fermentation pro le and higher alcohols experimental and predicted for run 31. CO2 pro le vmax (h1) Experimental Predicted Percentage error 0.4601 0.5346 13.9 Ethanol Yeth=C [l(100 l1)] 0.5111 0.5120 0.2 Substrate Ysubs=C (g l1) 4.0042 4.7702 16.1 n-Propanol Yalc=C (mg l1) 1.9995 2.3176 13.7 Iso-butanol Yalc= C (mg l1) 2.8056 2.4824 13.0 Iso-amyl alcohol Yalc= C (mg l1) 9.6397 10.3181 6.6

Table 5. Experimental and predicted yield coef cient parameters for esters: run 31. Ethyl acetate YEst= C,1 (mg l1) Experimental Predicted Percentage error 0.8124 0.9140 11.1 YEst=C,2 (mg l1) 2.8708 3.1662 9.3 Iso-butyl acetate YEst= C,1 (mg l1) 0.0000 0.0022 100 YEst=C,2 (mg l1) 0.0183 0.0101 81.2 Iso-amyl acetate YEst= C,1 (mg l1) 0.0000 0.0538 100 YEst=C,2 (mg l1) 0.3288 0.3011 9.2 Ethyl hexanoate YEst= C,1 (mg l1) 0.0140 0.0109 28.4 YEst= C,2 (mg l1) 0.0329 0.0271 21.4

coef cients and umax are compared in Tables 4 and 5. It is clear from the values in Tables 4 and 5 that in general there is reasonably good agreement between the two sets of yield coef cients, thus indicating that the model can be used to predict these parameters as a function of the process conditions considered here. Agreement between experimental and predicted values is not as close for yield coef cients with low absolute values; this may be as much a result of accuracies in experimental determination of these coef cients as with predictive values. It is important to note that a wider range of process conditions has been considered here than by previous researchers. It is dif cult to compare values of yield coef cients with values reported previously in the literature, because of differences in the process and conditions used here as compared to those from other studies. However, yield coef cients for ethanol, for example, are in relatively good agreement with those quoted previously (Titica et al., 2000). As described above, a longer-term objective is to develop models for prediction of fermentation behaviour at commercial scale. In a rst step towards this goal, the kinetic models developed here were used to evaluate fermentation performance at pilot scale. For the pilot scale data available, only CO2 evolution and wort density (substrate uptake) values

had been collected, i.e. there was no avour data. A CO2 evolution pro le was calculated for the pilot-scale process conditions, assuming solution homogeneity and using the models described above, for comparison with the measured evolution pro le, using the following procedure. Initially equation (7) [and yield coef cient equations for substrate and ethanol, which are needed to determine values for S and E in equation (7)] was integrated using values of umax, KI, KS (calculated as described above for laboratory scale as a function of process conditions) to give variation of Cp with time; dCp=dt could then be calculated. Using this equation, the CO2 evolution pro le, which would be expected for a small-scale (presumably homogenous) fermentation run at the same process conditions as used for the generation of 11 litre fermentation data was determined. This CO2 evolution pro le is compared with the pilot scale experimental data in Figure 1. An effectiveness factor (Z) was evaluated from this data as a function of time according to: (Cp)pilot=(Cp)laboratory; this is shown in Figure 2. Also shown in Figure 2 is the effectiveness factor calculated according to the following equation, presented by D az et al. (1996): Z b en (10) Trans IChemE, Vol 81, Part C, March 2003

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less effective than predicted based on an assumed homogenous 1 litre fermentation. However, from Figure 2 it is clear that the effectiveness factor predicted by equation (10) is not in close agreement with (Cp)pilot=(Cp)laboratory calculated for this case. However, it should be remembered that the constants used in equation (10) to calculate the effectiveness factor were determined for a much larger scale vessel than is under consideration here; ideally values for these constants should be determined as a function of scale, but much more large-scale data would be required to achieve this. The large discrepancy at shorter time intervals is somewhat misleading as it arises because of a time lag in the CO2 pro le predicted by the model; this could be explained if in the experimental case the wort was saturated with CO2 at the start of the fermentation. The agreement between experimental and predicted effectiveness is closer towards the end of the fermentation. DISCUSSIONS AND CONCLUSIONS
Figure 1. Variation of measured carbon dioxide production rate for the j ), compared with the 11 litre fermentation as a function of time ( rate predicted from the model equations for an homogeneous 1 litre fermentation ( ).

where b and n are constants, which were reported to be typically 0.4 and 0.2, respectively, for a Saccharomyces cerevisiae fermentation of 200 m3 (D az et al., 1996). e is the power input per unit volume which was calculated for the pilot-scale fermenter using the following equation: Hf 1 H P 2 e QCO2 g Hf r ln (11) V Hf
where QCO2 is the CO2 evolution rate, g is acceleration due to gravity, r is density of the uid, Hf is atmospheric pressure expressed in terms of meters of uid in the fermenter and H is the height of liquid in the fermenter. It is clear from Figure 1 that the larger scale fermentation is

Figure 2. Variation of effectiveness factor for the 11 litre fermentation, as a function of time, calculated (i) as (Cp)pilot=(Cp)laboratory (), and (ii) using equation (10) ().

A mathematical model has been developed for predicting yield coef cients for ethanol, substrate, esters and higher alcohols from on-line measurements of CO2 production rates, as a function of temperature, pitching rate, gravity and initial dissolved oxygen concentration in the wort, for 1 litre laboratory impeller agitated fermentations in which mixing is assumed to be homogenous. This model is based on an approach reported for fermentations agitated solely by CO2 evolution by previous researchers (Trelea et al., 2001; Titica et al., 2000), who primarily considered the effect of temperature, yeast inoculum and pressure on yield coef cients. These previous researchers considered the dependence on process conditions of yield coef cients for key avour compounds only, assuming that yield coef cients for substrate and ethanol were independent of process conditions; results here, however, clearly show that yield coef cients for ethanol and substrate do vary with process conditions investigated. This type of model should be valuable to the brewing industry as CO2 is a relatively easy and inexpensive parameter to measure continuously on-line. Such models could be used for optimization in terms of process conditions to achieve a speci ed set of yield coef cients. It is also suggested here that this kinetic model can be used to predict yield coef cients at larger scale, as a function of process conditions considered here, by means of the effectiveness factor approach proposed previously (D az et al., 1996). This approach has been used to compare CO2 evolution predicted for homogenous small-scale fermentation with measured values for an 11 litre scale fermentation. Demonstration of this approach for other scenarios, e.g. to predict yield coef cients as a function of process conditions at a range of scales, is limited by the current lack of availability of large-scale avour and CO2 data. It is suggested here that, as a rst stage towards developing models for beer production at larger scale, this approach may provide useful information regarding effect of changing process conditions and mixing input on yield coef cients for key avour components. Whilst it is appreciated that this approach is relatively simplistic, it has value in relying on measurable parameters. It is hoped that this basic demonstration of this approach will encourage collection of relevant large-scale process data to allow more extensive model validation. More complex computational

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HEPWORTH et al.
r Z umax wort density, g l1 effectiveness factor maximum speci c CO2 production rate, h1

models are likely in the long term to provide more accurate predictions; however, the development of such models, in integration with kinetic models, is far from trivial. Furthermore, methods for validation of such models would need to be developed. NOMENCLATURE
Alc C0 Cdc,m Cp D E Est g H Hf In KI KS P P QCO2 S S0 t T V YAlc= C YE= C YEst= C,1 YEst= C,2 YS= C X X0 Subscript n higher alcohol concentration, mg l1 initial CO2 production rate coef cient, l g1 CO2 concentration at maximum CO2 production rate, l l1 wort total volume of CO2 at time t per litre of wort, l l1 initial dissolved oxygen concentration in the wort, % air saturation ethanol concentration, l (100 l1) i.e. % v=v ester concentration, mg l1 acceleration due to gravity, m s2 height of uid in fermenter, m atmospheric pressure in terms of meters of uid in fermenter, m normalised factor value, varying between Imax and Imin product inhibition coef cient, (100 l)2 l2 substrate limitation coef cient, g l1 pitching rate, g l1 power input, W CO2 evolution rate, l l1 s1 substrate concentration, g l1 initial substrate concentration, g l1 time, h fermentation temperature, C volume, m3 yield coef cient for higher alcohols based on CO2 production, mg l1 yield coef cient for ethanol based on CO2 production, l (100 l1) yield coef cient for esters based on CO2 production for phase 1, mg l1 yield coef cient for esters based on CO2 production for phase 2, mg l1 yield coef cient for substrate based on CO2 production units, g l1 yeast concentration, g l1 initial yeast concentration, g l1 normalized factor value [according to equation (8)]

REFERENCES
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ACKNOWLEDGEMENTS
The authors would like to thank EPSRC for providing funding for this work. The agreement of the Chief Executive Of cer of BRi to the publication of this work is gratefully acknowledged.

ADDRESS
Correspondence concerning this paper should be addressed to Dr J. Varley, Department of Chemical Engineering and Chemical Technology, Imperial College London, South Kensington Campus, London SW7 2BY, UK. E-mail: j.varley@imperial.ac.uk The manuscript was received 8 November 2002 and accepted for publication after revision 6 March 2003.

Greek symbols e power input per unit volume, W m3 m speci c growth rate constant, h1

Trans IChemE, Vol 81, Part C, March 2003