A method for liquid fermentation of R. ginkgo crown, product and application thereof
A technology of L. coronoidans and liquid fermentation, which is applied in the directions of microorganism-based methods, applications, biochemical equipment and methods, etc., can solve the problem of less fermentation bacteria, achieve sensory properties improvement, increase added value, and enrich types. Effect
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[0050] Preparation of spore suspension: Scrape off the spores of S. coronis grown on PDA plane medium with an inoculation needle, transfer them into sterile water containing glass beads, shake at 28°C and 200rpm for 60min to break up the spore mass, and use a hemocytometer to disperse the spores. Count the spores on the counting board and adjust the concentration so that the concentration of the spore suspension is 1.7×10 8 cfu / mL (in the examples, the spore suspension was inoculated with this concentration).
[0051] Determination of the number of spores in the fermented broth: In a super-clean bench, use a pipette gun to draw 1mL of the fermented broth into a 100mL conical flask, add 9mL of sterile water, add a few glass beads to the conical flask, and put it in a 28 ℃, 200rpm shaker for 60min, take it out, and then dilute it 50 times, and count the spores with a hemocytometer under a microscope.
Embodiment 1
[0053] (1) Activating and separating S. coronoidis, and preparing spore suspension, the concentration of spore suspension is 1.7×10 8 cfu / mL;
[0054] (2) Ginkgo nuts are obtained by removing the bone middle testa and inner testa of the ginkgo, adding water to grind the pulp, adjusting the concentration of the ginkgo pulp to 8g (dry weight) / 100mL, taking 100mL of the ginkgo pulp and placing it in a 250mL Erlenmeyer flask , and add 0.5g of calcium chloride, 0.4g of magnesium sulfate, 0.125g of zinc sulfate and 0.063g of ferrous sulfate as a liquid medium, and sterilize at 115°C for 20 minutes for later use. The pH is natural and does not need to be adjusted;
[0055] (3) Inoculate 3mL of the spore suspension of Cystis coronoidis of step (1) in the liquid medium prepared in step (2) under aseptic conditions, after inoculation, at 160rpm, ferment for 3 days at 28°C, and take samples for determination every day The number of spores was obtained to obtain the liquid fermentation p...
Embodiment 2
[0057] (1) Activating and separating S. coronoidis, and preparing spore suspension, the concentration of spore suspension is 1.7×10 8 cfu / mL;
[0058] (2) Ginkgo nuts are obtained by removing the bone middle testa and inner testa of the ginkgo, adding water to grind the pulp, adjusting the concentration of the ginkgo pulp to 5g (dry weight) / 100mL, taking 150mL of the ginkgo pulp and placing it in a 250mL Erlenmeyer flask , and add 0.75g of calcium chloride, 0.6g of magnesium sulfate, 0.195g of zinc sulfate and 0.0945g of ferrous sulfate as a liquid medium, and sterilize at 115°C for 20 minutes for later use. The pH is natural and does not need to be adjusted;
[0059] (3) Inoculate 5 mL of the spore suspension of Cystis coronoidis of step (1) in the liquid medium prepared in step (2) under aseptic conditions, after the inoculation, at 160 rpm, ferment for 3 days at 28° C., and take samples for determination every day The number of spores was obtained to obtain the liquid ferm...
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