CN104004803A - Method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu - Google Patents
Method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu Download PDFInfo
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- CN104004803A CN104004803A CN201410218433.4A CN201410218433A CN104004803A CN 104004803 A CN104004803 A CN 104004803A CN 201410218433 A CN201410218433 A CN 201410218433A CN 104004803 A CN104004803 A CN 104004803A
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Abstract
The invention provides a method for producing antimicrobial peptide through fermentation of brevibacillus laterosporu. In fermentation of the brevibacillus laterosporu, when the yield of the antimicrobial peptide is no longer increased, active carbon is directly added into the fermentation liquor; as the active carbon adsorbs the fermentation product, the inhibiting effect of the product is relieved, and the density of the thallus and the yield of the antimicrobial peptide are greatly improved. According to the method, the active carbon is taken as an adsorption stripping coupling agent, has low cost, is environmentally friendly, and can be directly added in the fermentation process without being activated in advance; the antimicrobial peptide can be adsorbed without regulating the pH value of the fermentation liquor; and the method is simple and easy to promote, and the active carbon can be regenerated through desorption after fermentation, thus being recycled repeatedly, and lowering the production cost.
Description
Technical field
The present invention relates to microorganism fermentation field, specifically, relate to a kind of method of utilizing Brevibacillus laterosporus fermentative production antibacterial peptide.
Background technology
Along with the raising of people's living standard and the pay attention to day by day to food safety, human consumer requires to reduce the use of Chemical Preservative and antibiotic residual, therefore exploitation safety, efficient biological preservative become the important development direction of foodstuffs industry, wherein antibacterial peptide (Antimicrobial peptide, AMP) because its chemical nature is protein, drug residue free, non-immunogenicity, nontoxicity in human or animal body, have potential using value aspect instead of chemical sanitas and microbiotic.
Antibacterial peptide is a kind of micromolecule polypeptide being extensively present in organism; first separates the silkworm chrysalis that the antibacterial peptide obtaining comes from giant silkworm; up to now; in many animals, plant and microorganism, find the endogenic antibacterial peptide of kind more than 600; but animal source and antimicrobial peptide of plant origins source are limited; be difficult to accomplish scale production; and microbial source antibacterial peptide has, fermentation period is short, cost is low, be easy to cultivation, be easy to realize the advantages such as suitability for industrialized production, has become the important directions of antibacterial peptide research.Have the history of more than 50 year both at home and abroad about the research of microbial antibacterial peptide, most study be the bacteriocin of originating in lactic acid bacterium---Nisin, its foodstuffs industry that gone through can be used for, but the Nisin of originating in lactic acid bacterium has the problem such as narrow antimicrobial spectrum, poor stability mostly, therefore, further the non-milk-acid bacteria antibacterial peptide of development of new becomes a urgent and long-term task.
Bacillus (Bacillus) is because it has safe applicating history in foodstuffs industry, especially Brevibacillus laterosporus, the advantages such as that the antibacterial peptide of its generation has is safe, has a broad antifungal spectrum, good stability, noresidue, it is an important sources of non-bacteriocin lab, become the focus of research in recent years, be widely used in food, feed and control of crop disease field.In recent years the research of Brevibacillus laterosporus antibacterial peptide is found, produce in the process of antibacterial peptide at fermentation of bacillus, after reaching certain output, antibacterial peptide no longer increases, show a kind of product and suppress phenomenon, not only affect the growth of self thalline, also suppressed the further synthetic of antibacterial peptide simultaneously.Based on this, improve the output of antibacterial peptide, do not reach production object by the continuous fermentation process of supplemented medium merely, and should consider how to remove this restraining effect of antibacterial peptide, realize the bottleneck problem of high yield thereby fundamentally solve restriction antibacterial peptide.
The technology that microorganism fermentation and product separation are coupled is to solve in fermenting process aspect product inhibition effect remarkable, and easy to operate, this original position separate fermentation technology can optionally be isolated continuously thalline is had to inhibition or unsettled product from fermented liquid, both can make fermenting process carry out to the synthetic direction of product, can also reduce the effect of product natural degradation under complex environment.The original position isolation technique relating at present mainly comprises film fermentation method, electroosmose process, solvent extration and absorption method.From suitability for industrialized production angle, absorption method is low with its cost, selectivity is high, exchange capacity is large, simple to operate, be easy to the advantages such as automatically control, become the effective way that realizes the high fermentation yield of Brevibacillus laterosporus antibacterial peptide.
In recent years, investigator starts to explore the output of utilizing fermentation fractionation by adsorption coupled method to improve small-molecular peptides both at home and abroad.[the Xu Hao such as Wu Zhaoliang, Wu Zhaoliang, Yin Hao, Yan Jin. nisin technique is produced in the coupling of fermentation fractionation by adsorption. polymer material science and engineering, 2010,26 (10): 155-158] report, in the fermentative production of Nisin, add Zeo-karb D113 to carry out adsorbing coupled fermentation as sorbent material, while arriving fermentation termination, tiring of Nisin brought up to 5189IU/mL by original 3350IU/mL, improved 54.9%; But resin cost is high, and need to carry out activation treatment, complicated operation before dropping into fermented liquid.[the Pongtharangku T such as Pongtharangku, Demirci A.Online recovery of nisin during fermentation and its effect on nisin production in biofilm reactor.Applied Microbiology and Biotechnology, 2007, 74 (3): 555-562] utilize microorganism fermentation and membrane sepn and fractionation by adsorption coupled method to produce Nisin, the method make Nisin tire improve nearly 4 times, effect is remarkable, but this complex manufacturing, cost is higher, and the impact of fermented liquid pH value is large, be not suitable for suitability for industrialized production.Therefore, find zymotechnique with low cost, simple to operate and become problem demanding prompt solution.
Summary of the invention
The object of this invention is to provide a kind of method of utilizing the product inhibition in cheap fractionation by adsorption couplant releasing antibacterial peptide fermenting process and then improving cell density and yield of antibacterial peptides.
In order to realize the object of the invention, a kind of method of utilizing Brevibacillus laterosporus fermentative production antibacterial peptide of the present invention, in Brevibacillus laterosporus (Brevibacillus laterosporu) fermenting process, in the time that yield of antibacterial peptides no longer increases, in fermented liquid, directly add gac, absorption by gac to tunning, removes product inhibition, thereby improves cell density and yield of antibacterial peptides.
Directly add during the fermentation a kind of fractionation by adsorption couplant---gac, gac is without activation treatment in early stage, and without preparing adsorption column, fermented liquid is without regulating pH value, product inhibition be can remove, the fermentation density of thalline and the output of antibacterial peptide improved.
The Brevibacillus laterosporus relating in the present invention includes but not limited to B.laterosporu AS1.6343, B.laterosporu AS1.2827, B.laterosporu AS1.2739, B.laterosporu AS1.864, B.laterosporu ACCC06527, B.laterosporu ACCC05440, B.laterosporu ACCC10390 etc.
Utilize the concrete technology of Brevibacillus laterosporus fermentative production antibacterial peptide as follows:
1) seed liquor preparation: by the nutrient broth medium (NB substratum) of the inclined-plane of Brevibacillus laterosporus or glycerine pipe access sterilizing, culture temperature 30-37 DEG C, incubation time 12-24h, obtains primary seed solution; Then primary seed solution is pressed to 3-5v/v% inoculum size access nutrient broth medium, 30-37 DEG C, cultivates 6-18h, and obtaining bacteria concentration is 10
7-10
8the secondary seed solution of cfu/mL;
2) fermentation culture: to the fermention medium that packs fermentor tank volume 60-80% in fermentor tank into, the cooling rear access step 1 of sterilizing) secondary seed solution prepared, inoculum size 3-5v/v%; Fermentation condition is: leavening temperature 30-34 DEG C, rotating speed 200-400r/min, air flow 1:0.1-1:0.5v/v/min, dissolved oxygen 60-70%, incubation time 12-24h;
3) charcoal absorption: in the time that fermentation 12-24h yield of antibacterial peptides no longer increases, in fermented liquid, add gac, add Fermented Condensed substratum simultaneously, gac addition 1-4w/v%, Fermented Condensed substratum additional amount 1-3v/v%, continues fermentation 12-24h; Fermentation condition is with step 2).
Wherein, step 2) described in the formula of fermention medium be: carbon source 0.5-3.0w/v%, nitrogenous source 0.5-3.0w/v%, inorganic salt 0.005-1.0w/v%, tensio-active agent 0.1-1.0w/v%, adjust pH7.0-7.4, prepare with water.
Step 3) described in the formula of Fermented Condensed substratum be: carbon source 5.0-30.0w/v%, nitrogenous source 5.0-30.0w/v%, inorganic salt 0.05-10.0w/v%, tensio-active agent 1.0-10.0w/v%, adjust pH7.0-7.4, prepare with water.
In above-mentioned antibacterial peptide zymotechnique, available carbon source is selected from one or more in glucose, sucrose, Zulkovsky starch, dextrin, molasses or glycerine etc.
In above-mentioned antibacterial peptide zymotechnique, available nitrogenous source is selected from one or more in corn steep liquor, extractum carnis, peptone, yeast powder or bean cake powder etc.
In above-mentioned antibacterial peptide zymotechnique, available inorganic salt are selected from Ca
2+, Zn
2+, Mg
2+, Mn
2+, Fe
2+or Cu
2+one or more in salt etc.
In above-mentioned antibacterial peptide zymotechnique, available tensio-active agent is selected from one or more in Tween-20, Tween-40, Tween-60, Tween-80, Triton X-100, Triton X-114, Triton X-116, TX-4, TX-6, TX-7, TX-8, TX-9, TX-10, AEO-3, AEO-7 or AEO-9 etc.
The gac adding in above-mentioned antibacterial peptide zymotechnique is Powdered or granular bamboo charcoal, charcoal, fruit shell carbon or coconut husk charcoal.After charcoal absorption tunning, can, by means regeneration such as desorption, realize the recycle of gac.
Adopt above-mentioned antibacterial peptide zymotechnique, carry out fermentative production in fermentor tank, compared with not adding gac, viable count improves 2-4 the order of magnitude, reaches 10
10-10
12cfu/mL, yield of antibacterial peptides improves 50-130%, tires as 2200-3500AU/mL.
The present invention's fermentation strain used is Brevibacillus laterosporus, is called again bacillus laterosporus, and its meta-bolites has the function that suppresses various bacteria, fungi.In foodstuffs industry, be applied to producing functional foodstuff and food antiseptic is fresh-keeping.In fodder industry, be can be used for feeding micro-ecological preparation by Ministry of Agriculture's approval, be applied to the cultivation of broiler chicken, meat duck, pig and shrimp, there is high security, be a kind of potential probiotic bacterium.
The present invention's fractionation by adsorption couplant used is gac, with low cost, environmentally friendly, in fermenting process, directly adds, and without carrying out activating pretreatment, and fermented liquid is without regulating pH value, can realize the absorption of antibacterial peptide.Technique is simple, is easy to promote, and after fermentation ends, can process regenerated carbon by desorption, has realized the repeatedly recycling of gac, has reduced production cost.
Antibacterial peptide zymotechnique provided by the invention has been removed the restraining effect of product in fermenting process, and make to ferment thalline and yield of antibacterial peptides significantly improve.
Utilize the antibacterial peptide of Brevibacillus laterosporus fermentative production, its chemical nature is protein, noresidue, security is good, and stability is high, has a broad antifungal spectrum, various bacteria and fungi can be suppressed, the industries such as food, medicine, livestock and poultry cultivation, biological husbantry can be widely used in.
Embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that in embodiment, technique means used is well known to those skilled in the art, the raw materials used commercial goods that is.
The percentage sign " % " relating in the present invention, if not specified, refers to mass percent; But the per-cent of solution, unless otherwise specified, refers to the grams that contains solute in 100mL solution.
Embodiment 1 utilizes the technique of Brevibacillus laterosporus fermentative production antibacterial peptide
Concrete technology is as follows:
1, seed culture
To preserve in the NB substratum of glycerine pipe access sterilizing of B.laterosporu AS1.2827,32 DEG C, incubation time 24h; Press 5v/v% inoculum size access NB substratum again, cultivate 8h for 32 DEG C, obtaining bacteria concentration is 10
7the secondary seed solution of cfu/mL.
2,5L fermentor cultivation
The secondary seed solution obtaining in step 1 is equipped with in the 5L fermentor tank of 3L fermention medium by the access of 5v/v% inoculum size, and 32 DEG C, 300r/min, cultivates 15h under air flow 1:0.5v/v/min condition.
Wherein, fermention medium (w/v): glycerine 2.0%, bean cake powder 2.0%, CaCl
20.005%, Tween-200.5%, adjusts pH7.2, prepares with water.
3, add Activated Carbon Adsorption Separation couplant
In step 2, in the time that cultivation 15h secondary fermentation liquid antibacterial peptide content no longer increases, in fermented liquid, add 3% gac, add 3v/v% Fermented Condensed substratum simultaneously, continue fermentation culture 20h by fermentation condition in step 2.
Wherein, Fermented Condensed substratum is 10 times of concentrated solutions of fermention medium in step 2.
After fermentation ends, measure viable count and antibacterial peptide is tired, compared with not adding the control group of gac, viable count has improved 2 orders of magnitude, and yield of antibacterial peptides has improved 80%.
Embodiment 2 utilizes the technique of Brevibacillus laterosporus fermentative production antibacterial peptide
Concrete technology is as follows:
1, seed culture
To preserve in the NB substratum of inclined-plane access sterilizing of B.Laterosporu AS1.2738,34 DEG C, incubation time 24h, then press 3v/v% inoculum size access NB seed culture medium, and cultivate 12h for 34 DEG C, obtaining bacteria concentration is 10
8the secondary seed solution of cfu/mL.
2, fermentor cultivation
The secondary seed solution obtaining in step 1 is equipped with in the 5L fermentor tank of 4L fermention medium by the access of 8v/v% inoculum size, carries out fermentation culture.
Fermention medium (w/v): Zulkovsky starch 1.5%, yeast powder 1.5%, CaCl
20.15%, ZnCl
20.008%, MnCl
20.05%, Tween-202.0%, pH7.0, prepares with water.
Fermentation condition: 34 DEG C of temperature, air flow 1:0.5v/v/min, dissolved oxygen 60%, dissolved oxygen is associated with rotating speed, and rotating speed 200-400r/min cultivates 16h.
3, add Activated Carbon Adsorption Separation couplant
In step 2, add 1.5% gac when cultivating in the backward fermented liquid of 16h, add 2v/v% Fermented Condensed substratum simultaneously, continue fermentation culture 24h by fermentation condition in step 2.Fermented Condensed substratum is 10 times of concentrated solutions of fermention medium in step 2.
After fermentation ends, measure viable count and antibacterial peptide is tired, compared with not adding the control group of gac, viable count has improved 3 orders of magnitude, and yield of antibacterial peptides has improved nearly one times.
Embodiment 3 gacs add the impact of time on viable count and yield of antibacterial peptides
Taking Brevibacillus laterosporus (B.laterosporu) AS1.864 as fermentation strain, first the glycerine pipe of preserving bacterial strain is accessed to the NB substratum of sterilizing, 32 DEG C, cultivate 24h, press 3v/v% inoculum size access NB substratum, cultivate 12h for 32 DEG C, obtaining bacteria concentration is 10
8the secondary seed solution of cfu/mL.
Adopt 5L fermentor tank, be equipped with in the system of 3L fermention medium by the access of 5v/v% inoculum size, at 32 DEG C, air flow 1:0.5v/v/min, under rotating speed 300r/min condition, carry out fermentation culture, respectively at fermentation the the 12nd, 16,20,24h, add 2% powdered active carbon and 2v/v% concentrate feed substratum, then continue fermentation 24h; Not add gac in contrast, viable count and yield of antibacterial peptides are in table 1.After fermentation 16h, add gac effect the most obvious, viable count is raising 3 orders of magnitude nearly compared with control group, and antibacterial peptide is tired and improved 80%.
NB seed culture medium (w/v): peptone 1.0%, extractum carnis 0.3%, NaCl0.5%, adjust pH7.2, prepare with water.In 121 DEG C, high pressure steam sterilization 20min.
Fermention medium (w/v): glucose 1.5%, corn steep liquor 1.8%, CaCl
20.15%, ZnCl
20.008%, Tween-800.1%, adjusts pH7.0, prepares with water.
Fermented Condensed substratum: 10 times of concentrated solutions of fermention medium.
The impact that the table 1 gac time of adding tires on viable count and antibacterial peptide
Embodiment 4 impacts of gac addition on viable count and yield of antibacterial peptides
Taking Brevibacillus laterosporus (B.laterosporu) AS1.864 as fermentation strain, first carry out secondary seed solution preparation, seed culture medium is NB substratum, 32 DEG C of culture temperature, obtaining bacteria concentration is 10
8the secondary seed solution of cfu/mL.
Adopt 5L fermentor tank, press the inoculum size access 3L fermention medium of 5v/v%, at 32 DEG C, air flow 1:0.5v/v/min, rotating speed 300r/min, after fermentation 16h, adds respectively 1%, 2%, 3% and 4% powdered active carbon, all add 2v/v% Fermented Condensed substratum, continue fermentation 24h simultaneously; Not add gac in contrast, viable count and yield of antibacterial peptides are in table 2.While adding 2.0% powdered active carbon, the raising effect of cell density and yield of antibacterial peptides is the most obvious, and viable count improves 3 orders of magnitude compared with the control, reaches 10
11cfu/mL, antibacterial peptide is tired and is doubled.
Fermention medium (w/v): glucose 1.5%, corn steep liquor 1.8%, CaCl
20.15%, ZnCl
20.008%, Tween-800.1%, adjusts pH7.0, prepares with water.
Fermented Condensed substratum: 10 times of concentrated solutions of fermention medium.
The impact that table 2 gac addition is tired on viable count and antibacterial peptide
Embodiment 5 impacts of Fermented Condensed material substratum additional amount on viable count and yield of antibacterial peptides
Taking Brevibacillus laterosporus (B.laterosporu) AS1.864 as fermentation strain, first carry out secondary seed solution preparation, obtaining bacteria concentration is 10
8the secondary seed solution of cfu/mL; Adopt 5L fermentor tank, press 5v/v% inoculum size access 3L fermention medium, at 32 DEG C, air flow 1:0.5v/v/min, under rotating speed 300r/min condition, after fermentation 16h, add 2% powdered active carbon, add respectively the Fermented Condensed substratum of 1v/v%, 2v/v% and 3v/v% simultaneously, continue fermentation 24h, not add gac in contrast, viable count and yield of antibacterial peptides are in table 3.The Fermented Condensed substratum that adds 2v/v% is the most remarkable to the raising effect of tiring, and approximately improves 90%.
Fermention medium (w/v): glucose 1.5%, corn steep liquor 1.8%, CaCl
20.15%, ZnCl
20.008%, Tween-200.1%, adjusts pH7.0, prepares with water.
Fermented Condensed material substratum: 10 times of concentrated solutions of fermention medium.
The impact that table 3 Fermented Condensed material substratum addition is tired on viable count and antibacterial peptide
Embodiment 630L fermentor tank amplifies scale fermentation test
Taking Brevibacillus laterosporus (B.laterosporu) AS1.864 as fermentation strain, in 32 DEG C, NB substratum carries out secondary seed solution preparation, and obtaining bacteria concentration is 10
8the secondary seed solution of cfu/mL.
Adopt 30L fermentor tank, by 5v/v% inoculum size access 20L fermention medium, at 32 DEG C, air flow 1:0.5v/v/min, rotating speed 300r/min, adds 2% powdered active carbon and 2v/v% Fermented Condensed substratum after fermentation 15h, then continues fermentation 20h.
Fermention medium (w/v): glucose 1.5%, corn steep liquor 1.8%, MnCl
20.05%, ZnCl
20.005%, CaCl
20.15%, adjust pH7.0, prepare with water.
Fermented Condensed substratum: 10 times of concentrated solutions of fermention medium.
After fermentation ends, viable count reaches 5.0 × 10
11cfu/mL, the antibacterial peptide 2916AU/mL that tires.With do not add gac compared with, viable count increase by 3 orders of magnitude, yield of antibacterial peptides has improved nearly one times.
Embodiment 7 Brevibacillus laterosporus antibacterial peptide antimicrobial spectrum tests
Utilize gac fermentation fractionation by adsorption coupling method to obtain Brevibacillus laterosporus antibacterial peptide, adopt the antimicrobial spectrum of Oxford cup inhibition zone method research antibacterial peptide.
Table 4 result shows, this antibacterial peptide has certain restraining effect to multiple food source property and avian pathogenic bacterium, comprises gram-positive microorganism: Listeria monocytogenes, streptococcus aureus, swine streptococcus, bacillus cereus, subtilis, bacillus megaterium etc.Gram-negative bacteria: intestinal bacteria, salmonella typhi, shigella flexneri, Pseudomonas aeruginosa, Caused by Yersinia enterocolitica, Enterobacter sakazakii etc.Visible, the antibacterial peptide being produced by Brevibacillus laterosporus has broad application prospects in the anticorrosion and livestock and poultry nonreactive cultivation field of biological food.
The antimicrobial spectrum of table 4 antibacterial peptide
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (9)
1. one kind is utilized the method for Brevibacillus laterosporus fermentative production antibacterial peptide, it is characterized in that, in Brevibacillus laterosporus (Brevibacillus laterosporu) fermenting process, in the time that yield of antibacterial peptides no longer increases, in fermented liquid, directly add gac, absorption by gac to tunning, removes product inhibition, thereby improves cell density and yield of antibacterial peptides.
2. method according to claim 1, it is characterized in that, described Brevibacillus laterosporus includes but not limited to B.laterosporu AS1.6343, B.laterosporu AS1.2827, B.laterosporu AS1.2739, B.laterosporu AS1.864, B.laterosporu ACCC06527, B.laterosporu ACCC05440, B.laterosporu ACCC10390.
3. method according to claim 1, is characterized in that, comprises the following steps:
1) seed liquor preparation: by the nutrient broth medium of the inclined-plane of Brevibacillus laterosporus or glycerine pipe access sterilizing, culture temperature 30-37 DEG C, incubation time 12-24h, obtains primary seed solution; Then primary seed solution is pressed to 3-5v/v% inoculum size access nutrient broth medium, 30-37 DEG C, cultivates 6-18h, and obtaining bacteria concentration is 10
7-10
8the secondary seed solution of cfu/mL;
2) fermentation culture: to the fermention medium that packs fermentor tank volume 60-80% in fermentor tank into, the cooling rear access step 1 of sterilizing) secondary seed solution prepared, inoculum size 3-5v/v%; Fermentation condition is: leavening temperature 30-34 DEG C, rotating speed 200-400r/min, air flow 1:0.1-1:0.5v/v/min, dissolved oxygen 60-70%, incubation time 12-24h;
3) charcoal absorption: in the time that fermentation 12-24h yield of antibacterial peptides no longer increases, in fermented liquid, add gac, add Fermented Condensed substratum simultaneously, gac addition 1-4w/v%, Fermented Condensed substratum additional amount 1-3v/v%, continues fermentation 12-24h; Fermentation condition is with step 2);
Wherein, step 2) described in the formula of fermention medium be: carbon source 0.5-3.0w/v%, nitrogenous source 0.5-3.0w/v%, inorganic salt 0.005-1.0w/v%, tensio-active agent 0.1-1.0w/v%, adjust pH7.0-7.4, prepare with water;
Step 3) described in the formula of Fermented Condensed substratum be: carbon source 5.0-30.0w/v%, nitrogenous source 5.0-30.0w/v%, inorganic salt 0.05-10.0w/v%, tensio-active agent 1.0-10.0w/v%, adjust pH7.0-7.4, prepare with water.
4. method according to claim 3, it is characterized in that step 2) described in fermention medium and step 3) described in the carbon source that uses in Fermented Condensed substratum be selected from one or more in glucose, sucrose, Zulkovsky starch, dextrin, molasses or glycerine.
5. method according to claim 3, it is characterized in that step 2) described in fermention medium and step 3) described in the nitrogenous source that uses in Fermented Condensed substratum be selected from one or more in corn steep liquor, extractum carnis, peptone, yeast powder or bean cake powder.
6. method according to claim 3, is characterized in that step 2) described in fermention medium and step 3) described in the inorganic salt that use in Fermented Condensed substratum be selected from Ca
2+, Zn
2+, Mg
2+, Mn
2+, Fe
2+or Cu
2+one or more in salt.
7. method according to claim 3, it is characterized in that step 2) described in fermention medium and step 3) described in the tensio-active agent that uses in Fermented Condensed substratum be selected from one or more in Tween-20, Tween-40, Tween-60, Tween-80, Triton X-100, Triton X-114, Triton X-116, TX-4, TX-6, TX-7, TX-8, TX-9, TX-10, AEO-3, AEO-7 or AEO-9.
8. method according to claim 3, is characterized in that step 3) in use gac be Powdered or granular bamboo charcoal, charcoal, fruit shell carbon or coconut husk charcoal.
9. according to the method described in claim 1-8 any one, it is characterized in that, after charcoal absorption tunning, regenerate by desorption means, realize the recycle of gac.
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| CN110577910A (en) * | 2019-09-17 | 2019-12-17 | 南京农业大学 | Brevibacillus laterosporus, antibacterial lipopeptide and application of antibacterial lipopeptide in agriculture and food |
| CN110577910B (en) * | 2019-09-17 | 2022-02-01 | 南京农业大学 | Brevibacillus laterosporus, antibacterial lipopeptide and application of antibacterial lipopeptide in agriculture and food |
| CN111019621A (en) * | 2019-12-11 | 2020-04-17 | 中国海洋石油集团有限公司 | Blocking remover and preparation method thereof |
| CN111269865A (en) * | 2020-04-01 | 2020-06-12 | 北京工商大学 | Brevibacillus laterosporus strain S62-9 and application thereof |
| CN111269865B (en) * | 2020-04-01 | 2020-12-01 | 北京工商大学 | Brevibacillus laterosporus strain S62-9 and application thereof |
| CN112553276A (en) * | 2020-12-14 | 2021-03-26 | 华北制药集团新药研究开发有限责任公司 | Production process for producing antibacterial peptide by using brevibacillus laterosporus |
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