Method for shortening high-salt fermentation period of lactic acid bacteria by controlling pH of two phases
A fermentation cycle and lactic acid bacteria technology, applied in the field of microbial fermentation, can solve the problems of high osmotic pressure of fermentation liquid, prolonged lactic acid bacteria lag period, inhibition of lactic acid bacteria physiological metabolism, etc., and achieve the effect of increasing bacterial biomass and shortening fermentation time.
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example 1
[0015] Example 1 (control group 1): Tetragenococcus halophilus CGMCC 3792 was fermented at a constant pH of 6.5 ( figure 1 )
[0016] (1) Seed culture: The glycerol tube storage solution of Tetragenococcus halophilus preserved at -80°C is inoculated into the seed culture medium at 5% (v / v) inoculum, and incubated at 30°C for 24 hours. The seed culture medium is (w / v): glucose 0.5%, soy peptone 2%, magnesium sulfate heptahydrate 0.025%, ascorbic acid 0.05%, sodium chloride 0%, pH 6.5;
[0017] (2) Expanded cultivation of secondary seeds; take the above-mentioned activated seed solution, inoculate it in an expanded medium with an inoculum of 5% (v / v), and incubate at 30°C for 24 hours. The secondary seed medium is (w / v): glucose 0.5%, tryptone 0.5%, soy peptone 0.5%, beef extract 0.5%, yeast extract 0.25%, magnesium sulfate heptahydrate 0.025%, ascorbic acid 0.05%, β-glycerol Sodium phosphate 1.9%, sodium chloride 6%, pH is 6.5;
[0018] (3) Cultivation in a 5 L fermentor; take the ...
example 2
[0019] Example 2 (Control group 2): Tetragenococcus halophilus CGMCC 3792 was fermented at a constant pH of 6.0 ( figure 1 )
[0020] Other conditions are the same as in Example 1. Take the secondary seed culture solution and inoculate it in a 5 L fermentor for fermentation at 30°C with an inoculum of 5% (v / v). The 5 L fermenter contains 4 L of liquid, and the stirring speed is 150 r / min, and the pH is controlled. Is 6.0. Fermentation culture 96 hour Reach the stable phase, the OD of the bacteria 600 Value is stable at 3.05 .
example 3
[0021] Example 3 (experimental group): Tetragenococcus halophilus CGMCC 3792 was fermented under two-stage pH control conditions ( figure 1 )
[0022] Other conditions are the same as in Example 1. Take the secondary seed culture solution and inoculate it in a 5 L fermentor for fermentation at 30°C with an inoculum of 5% (v / v). The 5 L fermenter contains 4 L of liquid and the stirring speed is 150 r / min. Before fermentation Control the pH to 6.5 after 60 hours, and control the pH to 6.0 after 60 hours. Fermentation culture 84 hour Reach the stable period, OD 600 Value is stable at 3.05 . Compared with the constant pH 6.5 (control 1), the biomass increased 17.3% ; Compared with the constant pH 6.0 (control group 2), the time to enter the stable phase Shortened 12 hour .
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