Construction method of bacillus phosphorus relieving engineering bacterial strain

A technology of engineering strains and bacillus, applied in biochemical equipment and methods, bacteria, treatment of microorganisms with electricity/wave energy, etc., can solve the problems that agricultural and environmental microorganisms are not involved, and achieve large mutation range and mutagenesis efficiency High and enhanced decomposition effect

Inactive Publication Date: 2003-04-16
ZHEJIANG UNIV
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Problems solved by technology

[0019] Although ion beam mutagenesis has obtained many high-yielding engineered bacteria in industrial microbial breeding, it has hardly been involved in agricultural and environmental microorganisms.

Method used

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  • Construction method of bacillus phosphorus relieving engineering bacterial strain
  • Construction method of bacillus phosphorus relieving engineering bacterial strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Application N + Ion beam implantation induces mutation of Bacillus megaterium to obtain an engineered strain that efficiently decomposes lecithin. Activated Bacillus megaterium on the slant of beef extract peptone medium, cultured at 28-30°C for 36-48 hours, eluted with normal saline, collected spores by centrifugation, diluted and spread on sterile empty dishes, and placed them on a clean bench. Ventilation and drying for 0.5~1h; energy consumption is 15~25KeV, dosage is 40~150×10 14 ions / cm 2 N + In vacuum (vacuum degree is 2 × 10 -2 ~4.2×10 -3 Pa), carry out intermittent pulse injection under sterile conditions; then wash out the spores with physiological saline, after screening on NBRIY (containing lecithin) medium, after identification, an engineering strain that efficiently decomposes lecithin is obtained. The ability to decompose lecithin is increased by 18 to 25%.

Embodiment 2

[0064] Application H + Ion beam implantation induces the mutation of the silicate bacteria KNP414 strain to obtain an engineered strain that efficiently decomposes inorganic phosphorus, inorganic potassium and lecithin.

[0065] KNP414 was activated on nitrogen-free medium, cultured at 28-30 °C for 30-36 h, inoculated into liquid medium without capsular spores, cultured at 28-30 °C, 200-250 rpm for 48-56 h, and the spores were collected by centrifugation , diluted and spread on a sterile empty plate, naturally dried for 0.5-1h; the energy used is 15-25KeV, and the dose is 60-200×10 14 ions / cm 2 H + In vacuum (vacuum degree is 3 × 10 -2 ~5×10 -3 pa), intermittent pulse injection under sterile conditions; then eluted spores with normal saline, screened on NBRIY medium (containing lecithin), and then re-screened on NBRIP medium, after identification, 2 strains were obtained with high efficiency decomposition Engineering strains KNP41401 and KNP41402 for inorganic potassium, ...

Embodiment 3

[0067] Apply He + Ion beam implantation induces mutation of Bacillus subtilis to obtain an engineered strain that efficiently decomposes phytate phosphorus. Activated Bacillus subtilis on the slant of beef extract peptone medium, cultured at 28-30°C for 36-48 hours, eluted with phosphate buffer, collected spores by centrifugation, diluted and spread on sterile glass slides, clean bench Ventilation and drying for 0.5~1h; the energy used is 10~20KeV, and the dosage is 40~150×10 14 ions / cm 2 He + In vacuum (vacuum degree is 2 × 10 -2 ~5×10 -3 pa), intermittent pulse injection under sterile conditions; then eluted spores with phosphate buffer, screened on NBRIY (phytate phosphorus) medium, and after identification, an engineering strain that efficiently decomposes phytate phosphorus was obtained. The ability to decompose phytate phosphorus is increased by 25 to 35%.

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Abstract

An engineered dephosphorylating strain of bacillus is configured through slant culture in beef paste-pepton culture medium, eluting the activated bacteria with physiological saline or buffering phosphoric acid solution to obtain the bacillus suspension, centrifugal separation to collect the spores, diluting with physiological saline or buffering phosphoric acid solution, coating on aseptic glass,drying, implanting ion beams in the bacillus, and eluting.

Description

technical field [0001] The invention relates to a method for constructing a phosphorus-dissolving engineering bacterial strain of Bacillus. Background technique [0002] Today, the development of sustainable agriculture has become the consensus of all mankind. However, the long-term development of chemical agriculture, animal husbandry and industry has caused serious environmental pollution, among which organic phosphorus is one of the important pollution sources. Organophosphorus pesticides in agricultural production, phytate phosphorus in livestock excrement, and various organic phosphorus in industrial wastewater and domestic sewage have caused a huge load on the environment, restricting the development of agriculture and aquatic industries. Therefore, the elimination of organic phosphorus pollution has become a topic of concern to countries all over the world. [0003] Microbial degradation is an important way to eliminate organic phosphorus p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N13/00
Inventor 陈集双胡秀芳唐爱菊唐骏陈益锦
Owner ZHEJIANG UNIV
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