Heterotrophic nitrification-aerobic denitrification pseudomonas mendocina as well as culture and application thereof
A technology for aerobic denitrification and denitrification medium, which is applied to the field of Pseudomonas mendoza in heterotrophic nitrification-aerobic denitrification, can solve problems such as weak ability to remove ammonium nitrogen, achieve high removal rate, Good denitrification effect and the effect of reducing COD in wastewater
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Embodiment 1
[0033] Example 1: Isolation of Pseudomonas mendoza strains of heterotrophic nitrification-aerobic denitrification
[0034] The sample is activated sludge from Wenzhou Xipian Sewage Treatment Plant. A certain amount of activated sludge was inoculated into the enrichment medium (NH 4 Cl 1g, sodium citrate 5g, MgSO 4 ·7H 2 O 0.1g, K 2 HPO 4 0.5g, NaCl 0.2g, MnSO 4 4H 2 O 0.02g, FeSO 4 0.02g, H 2 O 1000 ml, pH 7.0) at 30°C at 160-180 rpm for 3-4 days for enrichment culture, after bacteria grow, transfer to a new enrichment medium to continue enrichment culture, and repeat this 4-5 times. Then spread the appropriately diluted bacterial solution on the separation medium (agar 18g·L -1 , other components are the same as the enrichment medium) and cultured at 30°C for 48 h, pick a single colony and transfer it to a new isolation medium for streaking and purification, until it is a pure culture by microscopic examination, and then transfer to the preservation medium (beef Cr...
Embodiment 2
[0042] Example 2: Identification of Heterotrophic Nitrification-Aerobic Denitrification Strains
[0043] The colonies of the strain WZUF22 were round, translucent, with rough surfaces, irregular edges, light yellow colonies after being cultured in the isolation medium for 48 hours. Lange-negative, unipolar flagella.
[0044] 16S rDNA was amplified using bacterial genomic DNA as a template, using a pair of universal primers: upstream primer (P1): 5'-AGAGTTTGATCCTGGTCAGAACGAACGCT-3', downstream primer (P6): 5'-TACGGCTACCTTGTTACGACTTCACCCC-3', and purification of PCR products And the sequencing was completed by Shanghai Bioengineering Co., Ltd., and the sequencing results were compared and analyzed by GeenBank Blast. with Pseudomonas in GeenBank ( Pseudomonas sp .) The 16SrDNA sequence has a high homology, and P. mendocina The homology is 99.4%. Using MEGA4.1 software, the phylogenetic tree of the 16S rDNA sequence of strain WZUF22 and related species was displayed by the a...
Embodiment 3
[0046] Example 3: Strain WZUF22 removes NH 4 + -N and NO 3 - -N characteristics
[0047] Using the single factor test method, the research strain WZUF22 was subjected to heterotrophic nitrification to remove NH 4 + -N and aerobic denitrification to remove NO 3 - -N characteristics.
[0048] The experimental process is as follows: the preserved strain WZUF22 (2.0ml frozen tube thawed bacterial solution) was inoculated into 200ml LB medium (10g beef extract, 10g peptone, 5g NaCl, 18g agar, H 2 O 1000 ml, pH 7.0) in a 500ml Erlenmeyer flask, cultured at 30°C, 150rpm for 24 h, centrifuged at 8000rpm for 10min to obtain bacterial cells, washed twice with sterile water to prepare OD 680 0.900~1.000 bacterial suspension;
[0049] Bacteria suspension was transferred to 100ml nitrification medium (NH 4 Cl, carbon source, MgSO 4 ·7H 2 O, K 2 HPO 4 , NaCl, MnSO 4 4H 2 O, FeSO 4 , H 2 O, pH 4~10.5) or denitrification medium (carbon source, KNO 3 , K 2 HPO 4 , FeSO ...
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