Preparation method of high-concentration microalgae living cell long-acting preservative
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A technology of living cells and preservatives, applied in the field of cell viability maintenance and improvement, can solve the problems of high cost, corruption, loss of nutrients in algal cells, etc., and achieve the effect of maintaining vitality
Active Publication Date: 2019-08-23
TIANJIN UNIV OF SCI & TECH
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[0004] (1) Microalgae are transported at low temperature. Low temperature usually refers to the temperature at 0-4°C. At this temperature, this species can transport 2×10 microalgae per ml. 10 Cell concentration, there is a problem of high cost, and the vigor of algae is significantly reduced, so it is not suitable for the situation where live algae regulate the water quality of aquaculture water
[0005] (2) Microalgae are transported at room temperature. Microalgae are transported at room temperature in two ways: light and dark. However, under light conditions, the concentration of algae cells is low, generally 10 per ml. 6 less than 10 cells per milliliter 7 The algae cell concentrate will be spoiled within 24 hours, the transportation efficiency is low, and the cost is high
[0011] 3. Patent (200410036386.8) "Preparation method of dry products of Monocystis algae and its products and applications". This method is convenient for transportation by concentrating, drying, vacuuming or nitrogen-filling the algae, but the algae cells in this patent have died, and the algae partial loss of nutrients in cells
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Embodiment 1
[0035] Step 1. Make the cell concentration 2×10 6 / ml of Rhodopseudomonas capsulate and Halomonas campisalis were respectively inoculated into glutamate medium, cultured at 30°C in a 30L fermenter, the volume of culture solution was 15L, and aeration cultured 48h, Rhodopseudomonas capsulatum was 3×10 10 cfu / ml, the density of Halomonas campisalis is 2.8×10 10 cfu / ml, take 10L of each sample, and mix to obtain 20L mixed bacterial liquid. Centrifuge at 5000g for 10 minutes to obtain about 100mL of bacterial slurry.
[0036] Step 2: Mix 100ml of sodium alginate solution with a concentration of 20g / L and 100ml of bacterial mud collected in step 1, stir evenly, then add 10g / L of calcium chloride solution, and stir at the same time, after adding about 400ml, the solution When the degree of turbidity no longer deepens, let it stand for 30 minutes, then pour it into a bag made of 1500-mesh filter cloth, and let it stand for 1 hour. The liquid in the bag no longer seeps out, and abo...
Embodiment 2
[0046] Steps 1 to 4 are the same as Example 1,
[0047] Step 5. Store the mixed algal solution at 15-30°C in the dark.
[0048] At 2 days, 8 days, 1 month, and 2 months after storage, the odor was observed, and the viability was determined by PI-FDA staining.
[0049] Table 2. Test results of freshwater chlorella at different storage times
[0050] Sampling time odor survival rate 0 days Algae natural earthy smell 98％ 2 days later Algae natural earthy smell 96％ 8 days later Algae natural earthy smell 95％ 1 month later Slightly odorous earthy algae 92％ 2 months later putrid smell 12％
[0051] During the preservation process of freshwater Chlorella, its cell viability decreased gradually with the prolongation of preservation time. After being stored for one month, the survival rate is still 92%; within two months, it has been putrefied and only about 12% of the algal cells are alive.
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Abstract
The invention discloses a preparation method of a high-concentration microalgae living cell long-acting preservative. The preparation method comprises the steps of fermenting and culturing rhodopseudomonas capsulate and halophila campisalis in a glutamate culture medium to obtain bacterial sludge, embedding the bacterial sludge into sodium alginate carrier particles, and continuously adsorbing nutrient matrix powder containing inorganic salts, vitamins, organic carbon sources and the like into the carrier particles to obtain photosynthetic bacterium carrier particle nutrient salt mixed particles, and carrying out spray drying and powder homogenization to obtain the high-concentration microalgae living cell long-acting preservative. The preservative is simple to use, only a certain amount of preservative powder needs to be added into the concentrated algae liquid, bacteria are increased in the embedded particles, and the generated metabolites can maintain the ecological balance of an algae concentrated solution. The method is simple and feasible, and the vigor of concentrated algae living cells can be maintained for 10-30 days in spring, summer, autumn and autumn without decay.
Description
technical field [0001] The invention relates to a method for maintaining and improving cell viability during long-term storage and transportation of a large amount of microalgae concentrate in hot season. Background technique [0002] Aquatic microalgae such as Chlorella, Dunaliella salina, Haematococcus pluvialis, etc., because microalgae themselves contain a lot of nutrients and pigment content, are the best aquatic food. At present, there are mainly microalgae living cells and microalgae dry powder. Microalgae living cells are rich in nutrients, but are prone to death and spoilage during transportation. The existing microalgae concentrates are usually transported by low-temperature refrigeration, which leads to problems such as high cost and low algal cell viability. [0003] At present, the transportation of bait algae mainly includes the following methods. [0004] (1) Low temperature transport of microalgae, low temperature usually refers to the temperature of 0-4 °...
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