Pseudomonas stutzeri and its culture, immobilization and use

A technology of Pseudomonas stutzeri and culture medium, which is applied in the fields of Pseudomonas stutzeri and its cultivation, immobilization and application to achieve the effects of reducing COD in wastewater, high removal rate and good denitrification effect

Inactive Publication Date: 2014-01-08
WENZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented describes two methods: one uses pseudomonaerothermophiles that are able to fix atmospheric nitrogen into ammonium ion or other forms during respiration; another involves preparing stable fixed pseudomicroorganisms called Staphylococcus spp., with specific properties such as optimal temperatures for both types of organism. These techniques allow for effective treatment of various pollution sources including industrial wastes, sewage sludge from septic tanks, municipal solid waste, etc.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficiency of removing nitro compounds such as organically bound nitroxyl radicals and nitrobiles during treatments on sewage systems due to the development of new methods involving enzymatic decomposition without relying upon conventional techniques like chemical reduction or physical absorption.

Method used

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  • Pseudomonas stutzeri and its culture, immobilization and use
  • Pseudomonas stutzeri and its culture, immobilization and use
  • Pseudomonas stutzeri and its culture, immobilization and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment one: the separation of aerobic denitrification bacterial strain

[0044] The sample is activated sludge from the West Pian Sewage Treatment Plant in Wenzhou, Zhejiang.

[0045] Quantitative activated sludge was inoculated on the enrichment medium (KNO 3 1g, sodium succinate 5g, KH 2 PO 4 1g, FeSO 4 ·7H 2 O 0.20g, MgSO 4 ·7H 2 O 0.10g, H 2 O 1000mL, pH 7.2-7.6) at 30°C at 160-180 rpm for 3-4 days, and after bacteria grow, transfer to a new enrichment medium to continue the enrichment culture, repeating 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 under microscope inspection, and then transfer to the preservation medium (yeast Cream 5g, peptone 10g, NaCl 10g, agar 18g, H 2...

Embodiment 2

[0051] Example 2: Identification of aerobic denitrification strains

[0052] The colonies of strain WZUF25 were round, translucent, rough in surface, irregular in edges, light yellow in color after being cultured in the isolation medium for 48 hours. Lange-negative, unipolar flagella.

[0053] 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 ( Pseudmonas sp .) The 16SrDNA sequence has a high homology, and P. stutzeri The homology is 99.9%. Using MEGA4.1 software, the phylogenetic tree of the 16S rDNA sequence of strain WZUF25 and related species was displayed by the adjacent junction method. figure ...

Embodiment 3

[0055] Example 3: Removal of NO by aerobic denitrification of strain WZUF25 3 --N characteristics

[0056] Aerobic denitrification removal of NO by the research strain WZUF25 by single factor test 3 - -N characteristics. The experiment process is as follows: inoculate the preserved strain (2.0mL frozen tube thawed bacteria solution) into 200mL LB medium (yeast extract 5g, peptone 10g, NaCl 10g, H 2 O 1000 mL, pH 7.0) in a 500mL Erlenmeyer flask, cultured at 30°C, 160rpm for 24h, centrifuged at 8000rpm for 10min to obtain the bacteria, washed twice with sterile distilled water, and prepared bacteria with sterile water Suspension ( OD 680 0.900~1.000).

[0057] Then the bacterial suspension was transferred to 100mL denitrification medium (carbon source, KNO 3 , K 2 HPO 4 1g, FeSO 4 ·7H 2 O 0.20g, MgSO 4 ·7H 2 O 0.10 g, H 2 O 1000mL, pH 4~10) in a 250mL Erlenmeyer flask, cultured at a certain temperature (20~45°C) at a certain speed (0~250rpm) for 24 h, centr...

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Abstract

The invention discloses pseudomonas stutzeri and its culture, immobilization and use. The disclosed pseudomonas stutzeri is pseudomonas stutzeri WZUF25, is preserved in the China general microbiological culture collection center and has the preservation number of CGMCC NO. 7685. The pseudomonas stutzeri has wide aerobic denitrification suitable pH and temperature ranges, has a high NO3<->-N and NO2<->-N removal rate and a high NH4<->-N assimilation capability, and can provide a culture source for synchronous nitrification and denitrification. The invention also provides immobilized pseudomonas stutzeri prepared by an active carbon-calcium alginate embedding method. The immobilized pseudomonas stutzeri has a good NO3<->-N removal capability and good stability.

Description

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Claims

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Application Information

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Owner WENZHOU UNIVERSITY
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