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Composite microbial symbiosis induction culture technology based on SOD (Superoxide Dismutase), microbial symbiosis culture agent and application of composite microbial symbiosis induction culture method and microbial symbiosis culture agent

A symbiotic culture and microorganism technology, applied in microorganism-based methods, microorganisms, methods of using microorganisms, etc., can solve the problems of large amount of strain, long process flow, poor environmental adaptability, etc., and achieves low preparation cost and wide application range. , the effect of strong adaptation

Inactive Publication Date: 2019-04-26
浙江黛君生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The composite utilization of existing microorganisms is mostly the direct mixing of several strains. The strains obtained in this way cannot exist stably, and it is easy to lead to the imbalance of the flora and the simplification of the strains. The product quality of this composite microbial agent is open to question
[0004] In addition, the current fermentation technology of symbiotic compound strains is complicated to operate, and the cost is high. At the same time, the process of strain compounding is relatively cumbersome. Because each strain has different requirements on nutrition and environment, it is difficult to control artificial interference, especially for oxygen demand and tolerance. Bacterial strains with different degrees of tolerance, the symbiotic culture of the prior art is limited to the co-growth of the same type of different species of microorganisms. At present, the artificial induction and symbiosis of aerobic bacteria, anaerobic bacteria and facultative anaerobic bacteria that require different oxygen environments Cultivation technology has not made a breakthrough
At the same time, the existing compound strains are poor in stability and require relatively stable breeding conditions; they have poor environmental adaptability and can only be used for strain induction and screening of environmental conditions. Simplification
In practical application, the treatment effect of single aerobic bacteria or single anaerobic bacteria is poor. Most of the existing technologies are aerobic bacteria and anaerobic bacteria, which are applied in turn, and multiple treatments are carried out. The process is long and the project cost is high.
The actual application effect of the existing compound microbial bacterial agent is not significant, and the amount of bacteria required is too large

Method used

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  • Composite microbial symbiosis induction culture technology based on SOD (Superoxide Dismutase), microbial symbiosis culture agent and application of composite microbial symbiosis induction culture method and microbial symbiosis culture agent
  • Composite microbial symbiosis induction culture technology based on SOD (Superoxide Dismutase), microbial symbiosis culture agent and application of composite microbial symbiosis induction culture method and microbial symbiosis culture agent
  • Composite microbial symbiosis induction culture technology based on SOD (Superoxide Dismutase), microbial symbiosis culture agent and application of composite microbial symbiosis induction culture method and microbial symbiosis culture agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Aerobic bacteria: Bacillus subtilis;

[0098] Anaerobic bacteria: Methanogens;

[0099] 1. Collect strains and enrich culture in the culture medium: aerobic bacteria use the first culture medium, 37 ° C, conventional culture in a biochemical incubator; anaerobic bacteria use the second culture medium and enter the mixed gas in the anaerobic workstation: 10% h 2 , 10% CO 2 , 80%N 2 . Culture for 2-3 days.

[0100] 2. Take the bacterial solution described in 1 and add sterile water to dilute to 10 4 cfu / ml, 10 6 cfu / ml, 10 8 cfu / ml. Take 0.1-0.2mL of each dilution to smear on a plate, and culture in 3 parallels for 2-3 days at 37°C. Pick different single colonies according to the morphological characteristics of the colony, and streak for purification.

[0101] 3. Pick the purified colony and culture it upside down for 3-4 days, scrape the bacteria, and do molecular biology and morphological identification.

[0102] 4. Add 0.005-0.05% SOD (20000IU / g) to the anae...

Embodiment 2

[0113] Facultative anaerobes: Bacillus amyloliquefaciens, Bacillus Velez, Bacillus cereus and Bacillus pumilus;

[0114] Anaerobic bacteria: denitrifying bacteria;

[0115]1. Collect strains, enrich culture in the medium: the first medium for facultative anaerobic bacteria, place it in a three-gas incubator, and pass in a mixed gas: 5% O 2 , 10% CO 2 , 80%N 2 ; Anaerobic bacteria pass into mixed gas in the anaerobic workstation with the second culture medium: 10%H 2 , 10% CO 2 , 80%N 2 . Incubate for 2-3 days.

[0116] 2. Take the bacterial solution described in 1 and add sterile water to dilute to 10 4 cfu / ml, 10 6 cfu / ml, 10 8 cfu / ml. Take 0.1-0.2mL of each dilution to smear on a plate, and culture in 3 parallels for 2-3 days at 37°C. Pick different single colonies according to the morphological characteristics of the colony, and streak for purification.

[0117] 3. Pick the purified colony and culture it upside down for 3-4 days, scrape the bacteria, and do molec...

Embodiment 3

[0132] Aerobic bacteria: Acetobacter;

[0133] Facultative anaerobes: Bacillus licheniformis;

[0134] Anaerobic bacteria: denitrifying bacteria;

[0135] 1. Collect strains and enrich culture in the culture medium: the first culture medium for aerobic bacteria, 37 ° C, conventional culture in a biochemical incubator; the first medium for facultative anaerobic bacteria, placed in a three-gas incubator, Inject mixed gas: 5% O 2 , 10% CO 2 , 80%N 2 ; Anaerobic bacteria pass into mixed gas in the anaerobic workstation with the second culture medium: 10%H 2 , 10% CO 2 , 80%N 2 . Culture for 2-3 days.

[0136] 2. Take the bacterial solution described in 1 and add sterile water to dilute to 10 4 cfu / ml, 10 6 cfu / ml, 10 8 cfu / ml. For each dilution, take 0.1-0.2mL to smear on a plate, and culture in 3 parallels for 2-3 days at 37°C. Pick different single colonies according to the morphological characteristics of the colony, and streak for purification.

[0137] 3. Pick th...

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Abstract

The invention relates to the field of microbial symbiosis culture, in particular to a composite microbial symbiosis induction culture technology based on SOD (Superoxide Dismutase), a microbial symbiosis culture agent and application of a composite microbial symbiosis induction culture method and the microbial symbiosis culture agent. The microbial symbiosis culture induction method provided by the invention classifies target strains according to the demand for oxygen content; firstly, the strains with similar oxygen demand are combined and cultured to obtain a plurality of strain groups, theinduction of symbiosis culture is started from the strains with the lowest oxygen demand; by ensuring the amount of bioactive superoxide dismutase in the system and gradually increasing the oxygen content in culture conditions, the adaptability of the system is improved; when the oxygen content reaches a certain concentration, the strain groups with similar oxygen demand are added, and then the adaptability of the system to higher oxygen content is improved by ensuring the amount of the superoxide dismutase in the system and gradually increasing the oxygen content in the culture conditions; byrepeating the operation until symbiosis culture of all strain groups is finished, the microbial symbiosis culture agent can be obtained.

Description

technical field [0001] The invention relates to the field of microbial symbiotic culture, in particular to a SOD-based compound microbial symbiotic induction culture technology, microbial symbiotic culture agent and its application. Background technique [0002] People's utilization of microorganisms has gone through the stages of natural mixed culture, pure culture to purposeful mixed culture (co-cultivation). In recent decades, people have gradually discovered that some biochemical processes require more than two kinds of microorganisms to proceed, and some substances require the co-cultivation of multiple microorganisms to produce. It is a hot issue and has become an important method to increase production or discover new substances. [0003] The composite utilization of existing microorganisms is mostly the direct mixing of several strains. The strains obtained in this way cannot exist stably, and it is easy to lead to the imbalance of the flora and the simplification o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16C12N1/36C12R1/02C12R1/125C12R1/01C12R1/225C12R1/07C12R1/085C12R1/25C12R1/72C12R1/10C12R1/38
CPCC12N1/16C12N1/20C12N1/36
Inventor 王代军
Owner 浙江黛君生物医药科技有限公司
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