Method for producing pectate lyase by two-section pH (potential of hydrogen) control
A pectinase and alkaline technology, which is applied in the field of biological enzyme preparation, can solve the problems of unsuitable for industrial production and application, low activity of alkaline pectinase, and high cost, so as to save the chemical degumming process, shorten the fermentation cycle, The effect of improving quality
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Embodiment 1
[0030] Bacillus subtilis CCTCC NO:M200038 (applicant deposited in China Center for Typical Cultures (CCTCC) on November 20, 2000, see ZL01106844.2 for related patents) was activated on an inclined plane, and the bacterial lawn was inoculated to the medium containing the seeds Cultivate in shake flasks, the culture conditions are: 300ml Erlenmeyer flask liquid volume is 50ml, fermentation temperature 37 DEG C, 200rpm shaking culture 8h. Inoculate the initial fermentation medium with an inoculum size of 10%, the pH is controlled at 5.6-6.0, the temperature is 33-34°C, and the ventilation volume is 2.5-4.5Lmin -1 . The starch in the feed medium was liquefied with mesophilic amylase, and the amount of enzyme added was 32.5U g -1 , the liquefaction temperature is 55°C, and the liquefaction time is 30min. After initial fermentation to 16 hours, add pretreated feeding medium according to 20% of the volume of the initial fermentation medium, control the pH at 5.1-5.5, control the te...
Embodiment 2
[0035] The fermentation method and conditions were the same as in Example 1 except for the following changes. The pH before feeding was controlled to be 6.0-6.4, and the pH was controlled to be 5.5-5.9 after feeding. Through experimental determination, the crude enzyme liquid obtained by fermentation has the highest alkaline pectinase activity of 743Uml -1 . Fermentation process such as figure 1 shown.
Embodiment 3
[0037] The fermentation method and conditions were the same as in Example 1 except for the following changes. Before feeding, the pH was controlled to be 6.4-6.8, and after feeding, the pH was controlled to be 5.9-6.3. According to the experiment, the highest alkaline pectinase activity of the fermented crude enzyme liquid is 660U ml -1 . Fermentation process such as figure 1 shown.
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