CN101173231A - High-density bdellovibrio swim body fermenting and culturing technique - Google Patents

High-density bdellovibrio swim body fermenting and culturing technique Download PDF

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CN101173231A
CN101173231A CNA200710031166XA CN200710031166A CN101173231A CN 101173231 A CN101173231 A CN 101173231A CN A200710031166X A CNA200710031166X A CN A200710031166XA CN 200710031166 A CN200710031166 A CN 200710031166A CN 101173231 A CN101173231 A CN 101173231A
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bdellovibrio
host bacterium
concentration
precipitation
nutrient solution
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CNA200710031166XA
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Chinese (zh)
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蔡俊鹏
王泽秀
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华南理工大学
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Abstract

The invention relates to fermentation culturing technology of high density Bdellovibrio telotroch, comprising the steps as following: firstly, culturing the host-germ shaking table and adding DNB liquid after centrifugating to obtain host-germ suspension; secondly; adding the host-germ suspension into the DNB liquid and then adding the Bdellovibrio to culture, at last adding DNB liquid after centrifugating to obtain Bdellovibrio concentrated solution; thirdly; adding sodium glutamate into DNB liquid and then adding the host-germ suspension, at last adding the Bdellovibrio concentrated solution to make the concentration 1 to 10<3>PFU/mL. The fermentation liquid of the Bdellovibrio with density of 10<8> to 10<14>PFU/mL can be obtained by culturing in a fermentation cylinder for four to six days and the concentrated solution of Bdellovibrio telotroch can be obtained through concentration by centrifuging. The invention has the advantages of low cost, high final concentration of Bdellovibrio and wide application range, and can be used not only in the control of water source pathogen and food pathogen, but also in fields of medical science, environmental pollution, agriculture, industry and military matters.

Description

The fermentation culture technology of high-density Bdellovibrio telotroch
Technical field
The invention belongs to field of fermentation engineering, specifically be meant a kind of fermentation culture technology that can be applicable to reduce even eliminate fully the high-density Bdellovibrio telotroch of harmful bacterium, not only aspect water, food-borne pathogens prevent and treat, and will have huge application and development prospect at aspects such as medical science, environmental pollution, agricultural, industry, military fields.
Background technology
Though adopting the means of biological control to eliminate pathogenic bacterium is research focuses of every field such as the worker of the world today, farming, doctor, the main means of eliminating pathogenic bacterium at present are still the method for various physics and/or chemistry, comprise various antibiotic uses.These methods all exist tangible drawback, and at first, microbiotic is abused in a large number, can cause the generation of some side effects, as the resistance of pathogenic bacteria, suppress profitable strain etc.Secondly, though the method for acid-alkali treatment can reach certain sterilization/sterilization effect, behind harmful bacterium formation microbial film, the effect of these methods is just suitable undesirable.At last, the method for physics can only be applied to a certain link of association area to the elimination effect of various pathogenic bacterium, and is difficult to expand to all links in whole field.Yet the method for biological control can have powerful superiority, very likely makes the processing of large-scale water in its disease control of serving daily life and the biological warfare and the attack of terrorism, food source contact scar etc.
Can the degrade characteristic of protectiveness extracellular polymkeric substance and cracking host cell of Bdellovibrio makes it have great application prospect.At first, the host range of Bdellovibrio is extensive, common pathogens such as animal, daily life all there is stronger scavenging(action), and to the cracking abilities of pathogenic bacterium obviously greater than non-pathogenic bacteria, the Bdellovibrio preparation is when removing pathogenic bacterium, then exception little to the harm of the probiotics in the environment, cultivate Bdellovibrio as the host bacterium with probiotics; Secondly, Bdellovibrio does not almost have host resistance as a kind of antimicrobial agent, can be to the pathogenic bacterium persistent splitting action of playing stably, it can effectively prevent producer reorganization and gene level transfer between autogene group and other bacterial genomes, and does not also have the too single-minded shortcoming of host that phage has; Once more, Bdellovibrio also can be resisted some toxic substance of occurring in nature, can the encode anti-resistance albumen of multiple toxic substance of its genome, and this indication Bdellovibrio might become new antibiotic or sterilizing agent.There are some researches show that Bdellovibrio can not survive in the eukaryotic cell, disclose thus that it is minimum to the harm of human body; At last, Bdellovibrio can be dead because of " hunger " in the environment that does not have the host bacterium to exist, and therefore, uses the Bdellovibrio preparation not have residue problem.
At present, a kind of method of production phage bdellovibro preparation has been proposed in No. 200610039346.8 applications for a patent for invention of China, the application of the Bdellovibrio preparation that this method is produced, though can solve the side-effect problem that causes with methods such as microbiotic at present, but its Bdellovibrio preparation density of producing is lower, cause usage quantity bigger, cost is higher, and be to cultivate Bdellovibrio as the host bacterium with probiotics, with the leech plastid or/and the form of telotroch mixture, exists assorted cingula as finished product goes into, effect lags behind, potential use range is narrow, the probiotics cracking ability is crossed shortcomings such as strong.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art part, a kind of fermentation culture technology of high-density Bdellovibrio telotroch is provided, and then produces a kind of highdensity Bdellovibrio telotroch preparation that can be widely used in water, food-borne pathogens control and medical science, environmental pollution, agricultural, industry, military field etc.
For achieving the above object, main technical schemes of the present invention is as follows: high-density Bdellovibrio telotroch fermentation culture technology, and the step and the processing condition thereof of this technology are as follows:
Step 1: the preparation of host bacterium suspension:
The host bacterium is inoculated in the conventional substratum (being nutrient broth medium), place 20~40 ℃ of shaking tables to cultivate 12~24h, make it be in logarithmic phase, nutrient solution is at 2~15 ℃, centrifugal 15~the 40min of 5000~8000rpm, keep the precipitation abandoning supernatant, add in the precipitation and its volume ratio is 0.05~2% rare nutrient broth DNB nutrient solution, promptly obtain concentration and reach 10 18~10 26The host bacterium suspension of CFU/mL, it is standby to place 2~15 ℃ of refrigerators to preserve then;
Step 2: the preparation of Bdellovibrio telotroch concentrated solution:
In rare nutrient broth DNB nutrient solution, add the host bacterium suspension of step 1 preparation earlier, make the concentration of host bacterium in DNB liquid reach 10 10~10 18CFU/mL, insert the Bdellovibrio spot of cultivating out with ordinary method again, then this nutrient solution is cultivated 20~60h at 20~40 ℃, again at 2~15 ℃, centrifugal 15~the 40min of 5000~8000rpm, keep supernatant liquor and discard precipitation, with supernatant liquor again at 2~15 ℃, the centrifugal 15~40min of 10000~20000rpm, keep the precipitation abandoning supernatant, be precipitated as the Bdellovibrio telotroch, add in the precipitation with its volume ratio be 0.05~2%DNB nutrient solution, mixing promptly obtains concentration and reaches 10 6~10 8PFU/mL Bdellovibrio telotroch concentrated solution, it is standby then this concentrated solution to be placed 2~15 ℃ of refrigerators to preserve;
Step 3: the fermentating culturing process of high-density Bdellovibrio telotroch:
Prepare a large amount of rare nutrient broth DNB nutrient solutions, and the monosodium glutamate solution of adding after the cellulose filter membrane filtration treatment, make the concentration of monosodium glutamate solution in nutrient solution reach 0.05~10mM, insert the host bacterium suspension that step 1 makes again, make host bacterium initial concentration reach 10 10~10 18CFU/mL inserts the Bdellovibrio telotroch concentrated solution that step 2 prepares then, makes the Bdellovibrio initial concentration in the mixed-culture medium reach 1~10 3Behind the pFU/mL, the fermentor tank of packing in 20~40 ℃ of fermentation culture of carrying out routine, adds the host bacterium twice at least in the culturing process, each 18~30h at interval, and guarantee that host's bacteria concentration reaches 10 in each nutrient solution 10~10 18CFU/mL, fermentation culture can obtain concentration and reach 10 after 4~6 days 8~10 14The Bdellovibrio fermented liquid of pFU/mL, at last that this fermented liquid is first at 2~15 ℃, centrifugal 15~the 40min of 5000~8000rpm, discard precipitation, keep supernatant liquor, supernatant liquor is again at 2~15 ℃, centrifugal 15~the 40min of 10000~20000rpm, keep the precipitation abandoning supernatant, add in the precipitation with its volume ratio be 0.05~2% rare nutrient broth DNB nutrient solution mixing, promptly obtain the Bdellovibrio telotroch concentrated solution that ferments.
Preferred processing condition is in the preparation of the described host bacterium of step 1 suspension: the host bacterium is inoculated in the nutrient broth medium, place 28~32 ℃ of shaking tables to cultivate 14~16h, make it be in logarithmic phase, nutrient solution is at 4~5 ℃, centrifugal 18~the 25min of 5500~6500rpm, keep the precipitation abandoning supernatant, add in the precipitation and its volume ratio is 0.1~0.5% rare nutrient broth DNB nutrient solution.
The fermentating culturing process of the described high-density Bdellovibrio of step 3 telotroch, preferred version are fermentation culture 5 days, and every interval 24h added the host bacterium one time in preceding 3 days, added altogether three times, make the concentration of host bacterium in nutrient solution maintain 10 10~10 18CFU/mL continues fermentation culture then to reaching 5 days, and promptly obtaining concentration is 10 8~10 14The Bdellovibrio fermented liquid of pFU/mL.
The optimum concn of Sodium Glutamate is 0.5~1mM in the described nutrient solution of step 3.
Described host bacterium is meant: can be by the various vibrios of Bdellovibrio cracked, intestinal bacteria, Aeromonas, Salmonellas, Pseudomonas aeruginosa, single false born of the same parents bacterium or Edward's mushroom pathogenic bacterium bacterial strain.
The present invention compared with prior art has following advantage:
1, the host bacterium described in this experiment is meant: various vibrios, intestinal bacteria, Aeromonas, Salmonellas, Pseudomonas aeruginosa class, pseudomonas, Edwardsiella or the like are a series of can be by Bdellovibrio cracked pathogenic bacterium bacterial strain.The host bacterium has guaranteed the lytic activity of Bdellovibrio as a kind of viable bacteria, improves prevention effect.Avoided the life-time service probiotics to cultivate Bdellovibrio and cause the latter to the lifting of probiotics cracking ability, to the real potential result who is harmful to the passivation of bacterium cracking ability.
2, the high-density Bdellovibrio telotroch preparation produced of the present invention is with a wide range of applications.For example, in experiment, when a large amount of board fallings of haliotis diversicolor Reeve seedling are dead, add this Bdellovibrio telotroch preparation the pathogenic bacterium elimination factor is reached 94.1%; In the experiment of the pathogenic bacterium of carrying, add this Bdellovibrio telotroch scavenging agent and can reach more than 99% the clearance rate of Vibrio parahaemolyticus at oyster, Penaeus vannamei and perch etc.; Isolating some Bdellovibrio of this experiment simultaneously for example BDZ-100, BDZ-103 etc. has pathogenic, as can to cause human generation food poisoning common bacteria at non-01 vibrio cholerae, vibrio alginolyticus, intestinal bacteria, Aeromonas, Salmonellas, Pseudomonas aeruginosa etc. to the mankind, its cleavage rate is illustrated the using value of Bdellovibrio at aspects such as food sanitation safes thus up to 88%, 83.3%, 90.8%, 89.2%, 95% and 90.7%.
3, avoid using the mixture of Bdellovibrio leech plastid or telotroch and leech plastid and make lag behind its action time in application, and avoid bringing into of other bacterium and limit shortcoming such as its potential use range.
4, among the present invention Bdellovibrio when carrying out fermentation culture, the initial density of Bdellovibrio is extremely low, help saving cost, and compare with other method, the concentration height of Bdellovibrio in the fermented liquid that after fermentation, obtains, after centrifugal concentrating, also will obtain the Bdellovibrio preparation of high density again, reduce the usage quantity of preparation in the reality.
5, both also mutant strain of laboratory strains, wild strain of the Bdellovibrio that present technique adopted, both from seawater, salt water, fresh water, land, also people source, animal source and plant-sourced.
6, the present invention as finished product, widens its range of application with telotroch greatly, makes it not only aspect water, food-borne pathogens prevent and treat, and all can use at aspects such as medical science, environmental pollution, agricultural, industry, military fields.
Embodiment
Embodiment 1
High-density Bdellovibrio telotroch fermentation culture technology, the step and the processing condition thereof of this technology are as follows:
Step 1: the preparation of host bacterium suspension:
Strain intestinal bacteria/the Pseudomonas aeruginosa that this laboratory is separated is inoculated in the nutrient broth medium as the host bacterium, place 20 ℃ of shaking tables to cultivate 14h, make it be in logarithmic phase, nutrient solution is at 4 ℃, the centrifugal 35min of 5000rpm, keep the precipitation abandoning supernatant, add in the precipitation and its volume ratio is 0.05% rare nutrient broth DNB nutrient solution, promptly obtain concentration and reach 10 18~10 26The host bacterium suspension of CFU/mL, it is standby to place 2~4 ℃ of refrigerators to preserve then;
Step 2: the preparation of Bdellovibrio telotroch concentrated solution:
In rare nutrient broth DNB nutrient solution, add the host bacterium suspension of step 1 preparation earlier, make the concentration of host bacterium in DNB liquid reach 10 12CFU/mL, insert the Bdellovibrio spot of cultivating out with ordinary method again, then this nutrient solution is cultivated 48h at 20 ℃, again at 4 ℃, the centrifugal 35min of 5000rpm, keep supernatant liquor and discard precipitation, with supernatant liquor again at 4 ℃, the centrifugal 35min of 10000rpm, keep the precipitation abandoning supernatant, be precipitated as the Bdellovibrio telotroch, add in the precipitation with its volume ratio be the 0.05%DNB nutrient solution, mixing promptly obtains concentration and reaches 10 6~10 8PFU/mL Bdellovibrio telotroch concentrated solution, it is standby then this concentrated solution to be placed 4 ℃ of refrigerators to preserve;
Step 3: the fermentating culturing process of high-density Bdellovibrio telotroch:
Prepare a large amount of DNB nutrient solutions, and add the monosodium glutamate solution after the cellulose filter membrane filtration treatment, make the concentration of monosodium glutamate solution in nutrient solution reach 0.5mM, insert the host bacterium suspension that step 1 makes again, make host bacterium initial concentration reach 10 12CFU/mL inserts the Bdellovibrio telotroch concentrated solution that step 2 prepares then, makes the Bdellovibrio initial concentration in the mixed-culture medium reach 10 3Behind the PFU/mL, with this mixed-culture medium 10L fermentor tank of packing into, carried out fermentation culture 5 days in 20 ℃, every interval 18h adds the host bacterium one time in preceding 3 days, makes the concentration of host bacterium in nutrient solution maintain 10 10~10 18CFU/mL continues fermentation culture then to reaching 5 days, and can obtain ultimate density is 10 8~10 14The Bdellovibrio fermented liquid of PFU/mL.At last that this fermented liquid is first at 4 ℃, the centrifugal 35min of 5000rpm, discard precipitation, keep supernatant liquor, supernatant liquor is again at 4 ℃, and the centrifugal 35min of 10000rpm keeps the precipitation abandoning supernatant, add in the precipitation with its volume ratio be 0.05% rare nutrient broth DNB nutrient solution mixing, promptly obtain Bdellovibrio telotroch fermentation concentrated solution.
Embodiment 2
High-density Bdellovibrio telotroch fermentation culture technology, the step and the processing condition thereof of this technology are as follows:
Step 1: the preparation of host bacterium suspension:
Strain Vibrio parahaemolyticus that this laboratory is separated or molten bath vibrios or Edwardsiella are inoculated in the nutrient broth medium as the host bacterium, place 26 ℃ of shaking tables to cultivate 18h, make it be in logarithmic phase, nutrient solution is at 10 ℃, the centrifugal 20min of 6000rpm, keep the precipitation abandoning supernatant, add in the precipitation and its volume ratio is 0.5% rare nutrient broth DNB nutrient solution, promptly obtain concentration and reach 10 18~10 26The host bacterium suspension of CFU/mL, it is standby to place 2~4 ℃ of refrigerators to preserve then;
Step 2: the preparation of Bdellovibrio telotroch concentrated solution:
In rare nutrient broth DNB nutrient solution, add the host bacterium suspension of step 1 preparation earlier, make the concentration of host bacterium in DNB liquid reach 10 14CFU/mL, insert the Bdellovibrio spot of cultivating out with ordinary method again, then this nutrient solution is cultivated 36h at 26 ℃, again at 10 ℃, the centrifugal 20min of 6000rpm, keep supernatant liquor and discard precipitation, with supernatant liquor again at 10 ℃, the centrifugal 20min of 16000rpm, keep the precipitation abandoning supernatant, be precipitated as the Bdellovibrio telotroch, add in the precipitation with its volume ratio be the 0.5%DNB nutrient solution, mixing promptly obtains concentration and reaches 10 6~10 8PFU/mL Bdellovibrio telotroch concentrated solution, it is standby then this concentrated solution to be placed 6~8 ℃ of refrigerators to preserve;
Step 3: the fermentating culturing process of high-density Bdellovibrio telotroch:
Prepare a large amount of DNB nutrient solutions, and add the monosodium glutamate solution after the cellulose filter membrane filtration treatment, make the concentration of monosodium glutamate solution in nutrient solution reach 5mM, insert the host bacterium suspension that step 1 makes again, make host bacterium initial concentration reach 10 14CFU/mL inserts the Bdellovibrio telotroch concentrated solution that step 2 prepares then, makes the Bdellovibrio initial concentration in the mixed-culture medium reach 10 2Behind the PFU/mL, with this mixed-culture medium 10L fermentor tank of packing into, carried out fermentation culture 5 days in 26 ℃, every interval 24h adds the host bacterium one time in preceding 3 days, makes the concentration of host bacterium in nutrient solution maintain 10 10~10 18CFU/mL continues fermentation culture then to reaching 5 days, and can obtain ultimate density is 10 8~10 14The Bdellovibrio fermented liquid of PFU/mL.At last that this fermented liquid is first at 10 ℃, the centrifugal 20min of 6000rpm, discard precipitation, keep supernatant liquor, supernatant liquor is again at 10 ℃, and the centrifugal 20min of 16000rpm keeps the precipitation abandoning supernatant, add in the precipitation with its volume ratio be 0.5% rare nutrient broth DNB nutrient solution mixing, promptly obtain Bdellovibrio telotroch fermentation concentrated solution.
Embodiment 3
High-density Bdellovibrio telotroch fermentation culture technology, the step and the processing condition thereof of this technology are as follows:
Step 1: the preparation of host bacterium suspension:
A strain Aeromonas or Salmonellas or pseudomonas that this laboratory is separated, be inoculated in the nutrient broth medium as the host bacterium, place 32 ℃ of shaking tables to cultivate 24h, make it be in logarithmic phase, nutrient solution is at 15 ℃, and the centrifugal 15min of 8000rpm keeps the precipitation abandoning supernatant, add in the precipitation with its volume ratio be 2% rare nutrient broth DNB nutrient solution, promptly obtain concentration and reach 10 18~10 26The host bacterium suspension of CFU/mL, it is standby to place 2~4 ℃ of refrigerators to preserve then;
Step 2: the preparation of Bdellovibrio telotroch concentrated solution:
In rare nutrient broth DNB nutrient solution, add the host bacterium suspension of step 1 preparation earlier, make the concentration of host bacterium in DNB liquid reach 10 18CFU/mL, insert the Bdellovibrio spot of cultivating out with ordinary method again, then this nutrient solution is cultivated 24h at 32 ℃, again at 15 ℃, the centrifugal 15min of 8000rpm, keep supernatant liquor and discard precipitation, with supernatant liquor again at 15 ℃, the centrifugal 15min of 20000rpm, keep the precipitation abandoning supernatant, be precipitated as the Bdellovibrio telotroch, add in the precipitation with its volume ratio be the 2%DNB nutrient solution, mixing promptly obtains concentration and reaches 10 6~10 8PFU/mL Bdellovibrio telotroch concentrated solution, it is standby then this concentrated solution to be placed 4 ℃ of refrigerators to preserve;
Step 3: the fermentating culturing process of high-density Bdellovibrio telotroch:
Prepare a large amount of DNB nutrient solutions, and add the monosodium glutamate solution after the cellulose filter membrane filtration treatment, make the concentration of Sodium Glutamate in nutrient solution reach 10mM, insert the host bacterium suspension that step 1 makes again, make host bacterium initial concentration reach 10 18CFU/mL, insert the Bdellovibrio telotroch concentrated solution of step 2 preparation then, after making Bdellovibrio initial concentration in the mixed-culture medium reach 10PFU/mL, with this mixed-culture medium 10L fermentor tank of packing into, carried out fermentation culture 5 days in 32 ℃, every interval 30h adds the host bacterium one time in preceding 4 days, makes the concentration of host bacterium in nutrient solution maintain 10 10~10 18CFU/mL continues fermentation culture then to reaching five days, and can obtain ultimate density is 10 8~10 14The Bdellovibrio fermented liquid of PFU/mL.At last that this fermented liquid is first at 15 ℃, the centrifugal 15min of 8000rpm, discard precipitation, keep supernatant liquor, supernatant liquor is again at 15 ℃, and the centrifugal 15min of 20000rpm keeps the precipitation abandoning supernatant, add in the precipitation with its volume ratio be 2% rare nutrient broth DNB nutrient solution mixing, promptly obtain Bdellovibrio telotroch fermentation concentrated solution.

Claims (5)

1. the fermentation culture technology of high-density Bdellovibrio telotroch, it is characterized in that: the step and the processing condition thereof of this technology are as follows:
Step 1: the preparation of host bacterium suspension:
The host bacterium is inoculated in the conventional substratum, place 20~40 ℃ of shaking tables to cultivate 12~24h, make it be in logarithmic phase, nutrient solution is at 2~15 ℃, centrifugal 15~the 40min of 5000~8000rpm, keep the precipitation abandoning supernatant, add in the precipitation and its volume ratio is 0.05~2% rare nutrient broth DNB nutrient solution, promptly obtain concentration and reach 10 18~10 26The host bacterium suspension of CFU/mL, it is standby to place 2~15 ℃ of refrigerators to preserve then;
Step 2: the preparation of Bdellovibrio telotroch concentrated solution:
In rare nutrient broth DNB nutrient solution, add the host bacterium suspension of step 1 preparation earlier, make the concentration of host bacterium in DNB liquid reach 10 10~10 18CFU/mL, insert the Bdellovibrio spot of cultivating out with ordinary method again, then this nutrient solution is cultivated 20~60h at 20~40 ℃, again at 2~15 ℃, centrifugal 15~the 40min of 5000~8000rpm, keep supernatant liquor and discard precipitation, with supernatant liquor again at 2~15 ℃, the centrifugal 15~40min of 10000~20000rpm, keep the precipitation abandoning supernatant, be precipitated as the Bdellovibrio telotroch, add in the precipitation with its volume ratio be 0.05~2%DNB nutrient solution, mixing promptly obtains concentration and reaches 10 6~10 8PFU/mL Bdellovibrio telotroch concentrated solution, it is standby then this concentrated solution to be placed 2~15 ℃ of refrigerators to preserve;
Step 3: the fermentating culturing process of high-density Bdellovibrio telotroch:
Prepare a large amount of rare nutrient broth DNB nutrient solutions, and the monosodium glutamate solution of adding after the cellulose filter membrane filtration treatment, make the concentration of monosodium glutamate solution in nutrient solution reach 0.05~10mM, insert the host bacterium suspension that step 1 makes again, make host bacterium initial concentration reach 10 10~10 18CFU/mL inserts the Bdellovibrio telotroch concentrated solution that step 2 prepares then, makes the Bdellovibrio initial concentration in the mixed-culture medium reach 1~10 3Behind the pFU/mL, the fermentor tank of packing into carries out conventional fermentation culture in 20~40 ℃, adds the host bacterium in the culturing process at least twice, each 18~30h at interval, and guarantee that host's bacteria concentration reaches 10 in each nutrient solution 10~10 18CFU/mL, fermentation culture can obtain concentration and reach 10 after 4~6 days 8~10 14The Bdellovibrio fermented liquid of pFU/mL, at last that this fermented liquid is first at 2~15 ℃, centrifugal 15~the 40min of 5000~8000rpm, discard precipitation, keep supernatant liquor, supernatant liquor is again at 2~15 ℃, centrifugal 15~the 40min of 10000~20000rpm, keep the precipitation abandoning supernatant, add in the precipitation with its volume ratio be 0.05~2% rare nutrient broth DNB nutrient solution mixing, promptly obtain the Bdellovibrio telotroch concentrated solution that ferments.
2. the fermentation culture technology of high-density Bdellovibrio telotroch according to claim 1 is characterized in that; Processing condition are in the preparation of the described host bacterium of step 1 suspension: the host bacterium is inoculated in the nutrient broth medium, place 28~32 ℃ of shaking tables to cultivate 14~16h, make it be in logarithmic phase, nutrient solution is at 4~5 ℃, centrifugal 18~the 25min of 5500~6500rpm, keep the precipitation abandoning supernatant, add in the precipitation and its volume ratio is 0.1~0.5% rare nutrient broth DNB nutrient solution.
3. the fermentation culture technology of high-density Bdellovibrio telotroch according to claim 1, it is characterized in that: the fermentating culturing process of the described high-density Bdellovibrio of step 3 telotroch, fermentation culture 5 days, every interval 24h added the host bacterium one time in preceding 3 days, add altogether three times, make the concentration of host bacterium in nutrient solution maintain 10 10~10 18CFU/mL continues fermentation culture then to reaching 5 days, and promptly obtaining ultimate density is 10 8~10 14The Bdellovibrio fermented liquid of PFU/mL.
4. according to the fermentation culture technology of claim 1 or 3 described high-density Bdellovibrio telotroches, it is characterized in that: the concentration of Sodium Glutamate is 0.5~1mM in the described nutrient solution of step 3.
5. according to the fermentation culture technology of claim 1 or 2 or 3 described high-density Bdellovibrio telotroches, it is characterized in that: described host bacterium is meant: can be by the various vibrios of Bdellovibrio cracked, intestinal bacteria, Aeromonas, Salmonellas, Pseudomonas aeruginosa, pseudomonas or Edward's mushroom pathogenic bacterium bacterial strain.
CNA200710031166XA 2007-10-30 2007-10-30 High-density bdellovibrio swim body fermenting and culturing technique CN101173231A (en)

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