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6H
2
O (0.020 g L
1
), CaCO
3
(0.020 g L
1
), and
concentrated HCl (0.13 ml L
1
); the pH was adjusted to 7.0 by adding NaOH.
3.2. Operational conditions
3.2.1. Inoculum build-up
Because xanthan gum constitutes the bacterial capsule, its production is growth-associated.
During inoculum buildup the aim is to increase the cell concentration but minimize the
production of xanthan because xanthan around the cells impedes mass transport of nutrients
and extends the lag phase of growth. Suppressing xanthan production while building up the
cell mass requires multiple stages of inoculum development (Silman and Rogovin, 1970;
Moraine and Rogovin, 1971; Cadmus et al., 1978; De Vuyst et al., 1987a; Peters et al., 1989;
Pons et al., 1989, 1990).
The microorganism is transferred from a complex solid culture medium (usually YM
agar) to a small volume (5 or 7 mL) of a complex liquid medium (usually YM), but the
incubation is limited to _7 h to prevent significant production of xanthan. This culture is
transferred to 40100 mL of a medium containing some inorganic salts (a semisynthetic
medium); the cells adapt to the new conditions that will be encountered in the production
phase (Santos, 1993). The inoculum volume for the production fermentor is between 5 and
10% of the total broth volume in the vessel. The number of stages needed to build up the
inoculum while suppressing xanthan synthesis, increases with the volume of the production
bioreactor (Santos, 1993).
3.2.2. Temperature
Temperatures employed for xanthan production range from 25 to 34C, but culture at 28C
and 30C is quite common (Table 4). The influence of temperature on xanthan gum
production has been widely studied. Moraine and Rogovin (1966) showed that 28C was
the optimal production temperature. Cadmus et al. (1978) concluded that a higher culture
temperature increases xanthan production but lowers its pyruvate content. Thonart et al.
(1985) reported an optimum process temperature of 33C, proposing a temperature of 25C
for growth and 30C for production. Shu and Yang (1990) claimed that the optimal
temperature depended on the final objective. For a high xanthan yield, a temperature between
31 and 33C was recommended, but culture at 2731C was better at attaining a high
pyruvate content in the gum. In addition, Shu and Yang (1990) concluded that the optimal
temperature for xanthan production depended on the production medium used.
F. Garc a-Ochoa et al. / Biotechnology Advances 18 (2000) 549579 557
A study of the influence of temperature (Garc a-Ochoa et al., 1997) using the earlier
reported optimized production medium (Garc a-Ochoa et al., 1992) showed that the optimal
production temperature for this medium was 28C, but production performance at 31C was
not much different (Fig. 5).
3.2.3. pH
Most authors agree that neutral pH is the optimum value for growth of X. campestris.
During xanthan production, the pH decreases from neutral pH to values close to 5 because of
acid groups present in xanthan. Some authors suggest that pH control is not necessary for this
Table 4
Operational conditions used in making xanthan gum in different bioreactors
Reference Bioreactor Temperature
(C)
pH Volume
(L)
Aeration rate
(L/ L min)
N (rpm)
Cadmus et al. (1978) Stirred tank 2030 6.8 (controlled) 10 1.5 225300
Rogovin et al. (1965) 28 7 227 0.5 90290
2268 30250
Moraine and
Rogovin (1966)
28 7 1 1000
Moraine and
Rogovin (1971)
28 7, 7.1
(controlled using
NH
4
OH)
8 1 5001000
Moraine and
Rogovin (1973)
28 7 (controlled)
Souw and
Demain (1980)
25 7 (controlled) 0.5 500
Pinches and
Pallent (1986)
30 7 (controlled) 10 0.4 600
2.5 1000
De Vuyst et al.
(1987a,b)
28 7 6 1 250700
Funahashi et al. (1987) 30 7 6 1 3501200
Peters et al. (1989) 28 7 (controlled) 0.3 200800
Shu and Yang (1990) 2034 7 (controlled) 1.16 800
Pons et al. (1990) 29 6.9 (controlled) 3.6 0.3, 0.6 500900
Kennedy et al. (1982) 30 7 (controlled) 3.5 0.5 400600
Schweikart and
Quinlan (1989)
26 7 (controlled) 1.2 1 3001300
Garc a-Ochoa et al.
(1997)
28 7 1.5 1 2101200
Pons et al. (1989) Bubble
column
29 6.9 (controlled) 13 1, 1.5
(constant)