Immobilized microbe pellet as well as preparation method and application thereof
A technology for immobilizing microorganisms and pellets, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of being easily affected by external factors, less application, time-consuming, etc., and achieve enhanced stability , Efficient removal, and the effect of preventing concentration reduction
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Embodiment 1
[0040] The strain in the immobilized microbial pellets of the present invention is Alcaligenes sp, which has been sent to the China Center for Typical Culture Collection on December 10, 2020 for preservation, the name is: Alcaligenessp.R3, and the preservation number is CCTCC NO: M2020882 Address: Wuhan University, Wuhan City, Hubei Province, China.
[0041] The described Alcaligenes sp.R3 is enriched from pig manure, separated and purified, and the specific steps are as follows:
[0042] Step 1: Enrichment, separation and purification of strains in immobilized microbial pellets
[0043] Pig manure samples were taken from a pig farm and stored in a -20°C refrigerator in a ziplock bag. Take 10g of the sample and put it into a Erlenmeyer flask containing glass beads, add 90mL of sterile water, shake at 170r / min, 30°C for 20min. Take the treated sample, draw 2mL supernatant and incubate in liquid medium for 2 days; then inoculate in screening medium containing 20mg / L tetracycli...
Embodiment 2
[0050] Preparation method of immobilized microbial pellets
[0051] S1, preparing the microbial strain into a bacterial suspension with a bacterial cell content of 1-9%;
[0052] The specific steps are as follows: Use an inoculation loop to pick the isolated and purified strains and inoculate them into LB liquid medium, place them in a constant temperature shaking incubator at 30°C and 140r / min for cultivation, and after cultivating for 12 hours, take a certain amount of culture solution in a centrifuge Centrifuge at 8000r / min in the tube for 10min, pour out the supernatant, wash with 0.9% normal saline repeatedly for 1-2 times, then dilute with normal saline, measure its absorbance at 600nm with a UV spectrophotometer with normal saline as a reference , and the bacterial solution when the OD600 nm was 1.0 was used as the bacterial suspension, and placed in a 4°C refrigerator for later use.
[0053] S2. Prepare a solution containing sodium carboxymethylcellulose and polydopam...
Embodiment 3
[0058] Determination of sodium carboxymethylcellulose content in immobilized microbial pellets.
[0059] The difficulty of making the immobilized microbial pellets is based on the drop into a spherical shape; calculate the integrity rate of the immobilized microbial pellets, the higher the integrity rate, the greater the mechanical strength of the immobilized microbial pellets, and the integrity rate = complete immobilization The number of immobilized microbial pellets / total number of immobilized microbial pellets×100%. Evaluation of the mechanical strength of the immobilized microbial pellets: 25 prepared immobilized microbial pellets were added to a 250mL Erlenmeyer flask filled with 50mL of water, and stirred at 1200r / min for 1min with a magnetic stirrer. Get 1.5%, 2%, 2.5%, 3%, 3.5% sodium carboxymethylcellulose and be dissolved in the ultrapure water of 100mL, prepare immobilized microorganism pellet according to the method for embodiment 2, pellet according to above-ment...
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