How is pyrroloquinoline quinone prepared?
High cell density fermentation (HCDF) has been developed in the last decade or two. It is a new type of fermentation technology that uses certain culture techniques and equipment to increase the ratio of bacterial biomass to target products and obtain high metabolites.
High cell density fermentation is currently reported in the following application areas: first, the use of engineered bacteria to produce various interferons, growth factors, biological enzymes, peptides, polysaccharides and other active ingredients; second, the application of high cell density fermentation technology in the food industry For the cultivation of living cells; the third is for yeast production of alcohol and the use of spirulina cells to produce spirulina. Due to the limitation of fermentation technology and equipment, there is no report on the industrial production of PQQ yet.
Steps to improve PQQ productivity:
CN201810866397.0 provides a method for high-density fermentation to increase the yield of PQQ, which shortens the fermentation period and improves the yield of PQQ. The method for increasing the yield of PQQ by high-density fermentation includes the following steps:
(1) Strain activation: Connect the hyphomicrobial strains to the slant culture medium, and obtain mature slant strains after cultivation;
(2) Preparation of seed mother liquor: the mature slant strain prepared in step (1) is inoculated into a seed culture medium and cultured on a shaker to obtain seed mother liquor;
(3) Seed pot culture: Use a 1.5-2m seed pot, add pure water to 50-60% of the volume of the seed pot, calculate the required seed pot culture material and add all the seed pot culture materials except anhydrous methanol while stirring. After the raw materials are completely dissolved and evenly dispersed, the volume is adjusted to 60-65% of the volume of the seed tank with purified water, mixed evenly, and the pH is adjusted to 7.0. After the seed tank is sterilized, it is cooled to 25-35°C, and the sterile filtered water is added. Mix evenly with water and methanol. Put the seed mother liquor obtained in step (2) into the seed tank, control the tank pressure at 0.08-0.10MPa, the tank temperature at 30-35°C, the air flow rate at 0.8-1.0VVM, and the pH at 6.8-7.0. Ventilate and stir for 42-50 hours to form a seed tank culture solution;
The seed pot culture material added to every 1.5m seed pot includes the following weights of raw materials:
(4) The initial preparation of the fermenter: use a fermenter of 20m or more, add purified water to 30-40% of the volume of the fermenter, add the fermenter culture material while stirring, and after the raw materials are completely dissolved and dispersed, use pure water to make the volume to 40-50% of the volume of the fermentor, mix well, adjust the pH to 7.0, and cool the fermenter to 25-35℃ after sterilization, and use sterile air to fill the seed tank obtained in step (3) with the inoculation ratio of 10-20% Press the culture broth from the seed tank into the fermentor, control the initial pressure of the fermenter to be 0.08-0.10MPa, the initial flow rate is 0.8-1VVM, the initial stirring speed is 110-130rpm/min, and the initial temperature is 30℃; bring it from the seed tank into the fermentor Unused anhydrous methanol is used as the initial concentration of methanol in the fermentation tank;
The fermenter culture material added to every 20m fermentor includes the following weights of raw materials:
(5) Fermentation and cultivation and feeding: before fermentation, add purified water and glycerin to the feeding tank, add sterile filtered anhydrous methanol after sterilization, and mix evenly to form feeding; during the fermentation process, when in the fermentation tank When the methanol concentration drops to 1000-1500ppm, start feeding to keep the total concentration of methanol and glycerin in the fermentor at 1000-1500ppm. Fermentation and cultivation are carried out for 5-6 days while feeding to obtain a high-content fermentation broth of PPQ.
The above is the introduction of the method steps for high-density fermentation to increase the yield of PQQ. PQQ can improve the metabolism and growth function of biological tissues and improve the immune function of the human body, which has high value. As a PQQ supplier, Clent Chemical has rich production experience and a professional production team, and the product quality is guaranteed. If you want to know more about PQQ, please continue to pay attention to our website.