CN105018398B - A kind of bacillus licheniformis culture medium and preparation method thereof - Google Patents
A kind of bacillus licheniformis culture medium and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of bacillus licheniformis culture medium and preparation method thereof, wherein, it is made by the raw material of following parts by weight：0.1 0.2 parts of 49 parts of corn pulp, 38 parts of peptone, 59 parts of sugarcane, 20 30 parts of steamed beef soup, 39 parts of roundpod jute seed, 4 10 parts of rhizoma cyperi, 59 parts of eleusine indica, 39 parts of astragalus, 4 10 parts of Chinese lizardtail rhizome, 0.1 0.3 parts of ammonium chloride, 0.05 0.2 parts of calcium bicarbonate, 0.05 0.3 parts of phosphoric acid hydrogen zinc and niter cake.And disclose preparation method.The beneficial effects of the invention are as follows：The culture medium gemma yield that the present invention obtains is high, at low cost, and obtained bacillus licheniformis bacteriostatic activity is high.
Description
Technical field
The present invention relates to culture medium field, more particularly to a kind of bacillus licheniformis culture medium and preparation method thereof.
Background technology
In rod-shaped, Dan Sheng, adjustable flora imbalance reaches therapeutic purposes, can promote for Bacillus licheniformis cell form and arrangement
Body is made to generate antibacterial substance, kill pathogenic bacteria.Resistant activity material can be generated, and takes oxygen effect machine by force with unique biology
System can inhibit the growth and breeding of pathogenic bacteria.
Value application：1. the effectively prevention diseases such as aquatic livestock enteritis, gill rot.2. it decomposes poisonous and harmful in aquaculture pond
Substance purifies water.3. with stronger protease, lipase, the activity of amylase, nutrient in feed is promoted to degrade, is made
Aquatic product animal absorbs more fully to feed.4. stimulating the development of aquatic livestock immune organ, enhance immunity of organism
Power.5th, promote the growth of normal physiological anaerobic bacteria in enteron aisle, adjust intestinal bacilli illness, recover function of intestinal canal.6th, to enteron aisle
Bacterium infection has special efficacy, and to light-duty or heavy chordapsus, light-duty and medium-sized acute bacillary dysentery etc. has obvious curative effects.
7th, resistant activity material can be generated, and there is unique biology take oxygen mechanism, the growth and breeding of pathogenic bacteria can be inhibited.
At present, bacillus licheniformis culture medium, there are low output, it is of high cost the problem of, and how to improve the ground of generation
The bacteriostatic activity of clothing bacillus.These problems are problems to be solved of the present invention.
The content of the invention
It is an object of the invention to provide a kind of culture mediums of bacillus licheniformis and preparation method thereof, obtain gemma yield
Culture medium high, at low cost, the bacillus licheniformis bacteriostatic activity obtained by basal culture medium culture are high.
In order to achieve the above object, the present invention provides a kind of bacillus licheniformis culture medium, wherein, by following parts by weight
Several raw materials are made：4-9 parts of corn pulp, 3-8 parts of peptone, 5-9 parts of sugarcane, 20-30 parts of steamed beef soup, 3-9 parts of roundpod jute seed, rhizoma cyperi
4-10 parts, 5-9 parts of eleusine indica, 3-9 parts of astragalus, 4-10 parts of Chinese lizardtail rhizome, 0.1-0.3 parts of ammonium chloride, calcium bicarbonate 0.05-0.2
0.1-0.2 parts of part, 0.05-0.3 parts of phosphoric acid hydrogen zinc and niter cake.
Wherein, it is made by the raw material of following parts by weight：6-8 parts of corn pulp, 4-7 parts of peptone, 6-9 parts of sugarcane, beef
25-30 parts of soup, 5-7 parts of roundpod jute seed, 6-8 parts of rhizoma cyperi, 6-9 parts of eleusine indica, 5-7 parts of astragalus, 6-8 parts of Chinese lizardtail rhizome, ammonium chloride
0.15-0.2 parts of 0.15-0.2 parts, 0.1-0.15 parts of calcium bicarbonate, 0.1-0.2 parts of phosphoric acid hydrogen zinc and niter cake.
Wherein, it is made by the raw material of following parts by weight：8 parts of corn pulp, 4 parts of peptone, 8 parts of sugarcane, 24 parts of steamed beef soup,
8 parts of roundpod jute seed, 8 parts of rhizoma cyperi, 6 parts of eleusine indica, 4 parts of astragalus, 5 parts of Chinese lizardtail rhizome, 0.15 part of ammonium chloride, 0.1 part of calcium bicarbonate, phosphorus
0.15 part of 0.2 part of sour hydrogen zinc and niter cake.
Wherein, it is made by the raw material of following parts by weight：7 parts of corn pulp, 5 parts of peptone, 7 parts of sugarcane, 27 parts of steamed beef soup,
6 parts of roundpod jute seed, 7 parts of rhizoma cyperi, 7 parts of eleusine indica, 6 parts of astragalus, 7 parts of Chinese lizardtail rhizome, 0.18 part of ammonium chloride, 0.13 part of calcium bicarbonate,
0.18 part of 0.15 part of phosphoric acid hydrogen zinc and niter cake.
In order to achieve the above object, the present invention also provides the preparation method of the bacillus licheniformis culture medium, wherein,
Its preparation method is：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in its total amount 1-3
The water of times amount impregnate 2-4 it is small when after carry out boiling 30 minutes -1 it is small when, filter the dregs of a decoction, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains, stir and be heated to 50- by second step
80 DEG C, continuous heating stir 30 minutes -1 it is small when, obtain mixed liquid；
3rd step, the NaOH solution that 70-80% is added in the mixed liquid obtained to second step adjust PH to 7.1-7.3, Ran Hou
110-130 DEG C, sterilize 15-18 minutes under the conditions of 110-120kPa, obtain bacillus licheniformis culture medium.
Sugarcane：【Source】For the stalk of grass sugarcane.It harvests after autumn, cuts and take aerial part.Prune top tip leaf
It ties up, air-dries and dry out.【Nature and flavor】It is cold, it is sweet.【Function】It is moisturized in, clearing heat and relieving fidgetness.
Roundpod jute seed：【Source】For the seed of Tiliaceae plant jute.【Chemical analysis】Seed glucoside containing corchorus olitorius, it is yellow
Pockmarks glucoside A, B, glucose erysimoside, the yellowish-white neat glucoside of sugar, roundpod jute seed alcohol glucoside A, roundpod jute seed poison glucoside B, C, glucosyl group（1→6）- long
Capsule roundpod jute seed glucoside, strophanthin is without also containing sucrose, gossypose, stachyose and verbascose.【Nature and flavor】It is bitter；Temperature.【Channel tropism】
Lung；Liver；Kidney channel.【Major function】Promoting blood circulation；Menstruation regulating；Cough-relieving.
Rhizoma cyperi：【Source】This product is the dry rhizome of sedge nutgrass flatsedge.Autumn excavates, and sing the hair palpus, after burning directly
It dries.【Chemical analysis】Containing glucose 8.3~9.1%, fructose 1.0~1.7%, starch 40~41.1%, volatile oil 0.65~
1.4%.【Nature and flavor】Pungent slight bitter sweet and neutral.【Major function】Regulating the flow of vital energy Xie Yu relieves pain menstruation regulating.
Eleusine indica：【Source】For the herb of grass family yard grass platymiscium eleusine indica.Summer and autumn are adopted, clean dry it is spare.
【Chemical analysis】Containing homo-orientin, cyanidenon -7-O- rutinosides, tricin（tricin）, 5,7- dihydroxy -3', 4',
5'- trimethoxy flavones, cyanidenon -7-O- glycosides, special Jing Su（vitexin）, isovitexin, heartsease glycosides displayed and
3-O- β-D- glucopyranosyls-cupreol and 6'-O- palmityls -3-O- β-glucopyranosyl-cupreol.【Property
Taste】It is sweet；It is light；It is cool.【Major function】Clearing heat and promoting diuresis；Removing pattogenic heat from the blood and toxic material from the body.
Astragalus：【Source】For the dry root of legume astragalus.Autumn excavates.Cleared soil cuts root head and branch
Root dries.【Chemical analysis】Astragalus containing sucrose, glucuronic acid, mucilaginous substance, several amino acid, bitter principle, choline, glycine betaine,
Folic acid, 2 ', 4 '-dihydroxy -5,6- dimethoxy isoflavan and kumatakenin.【Nature and flavor】It is sweet, tepor.【Major function】Benefit is defended solid
Table, inducing diuresis for removing edema, eliminating toxic, myogenic.
Chinese lizardtail rhizome：【Source】For the rhizome of Saururaceae plant saururus chinensis.7~September excavates underground rhizome, removes mud
Soil is put and is impregnated in hot water several minutes, and taking-up is dried.【Chemical analysis】Containing amino acid, organic acid, carbohydrate and condensed tannin.
【Nature and flavor】It is sweet；It is pungent；It is cold.【Major function】Li Shui dehumidifies；It is clearing heat and detoxicating.
The advantageous effect that technical scheme is brought is：The present invention can obtain gemma yield height, training at low cost
Base is supported, the bacillus licheniformis bacteriostatic activity that the medium culture by the present invention obtains is high.
Specific embodiment
In order to achieve the above object, the present invention is specifically described by following embodiment.
Embodiment 1
Formula：Corn pulp 4g, peptone 3g, sugarcane 5g, steamed beef soup 20g, roundpod jute seed 3g, rhizoma cyperi 4g, eleusine indica 5g, astragalus
3g, Chinese lizardtail rhizome 4g, ammonium chloride 0.1g, calcium bicarbonate 0.05g, phosphoric acid hydrogen zinc 0.05g and niter cake 0.1g.
Preparation process：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 1 times of its total amount
Boiling 30 minutes is carried out after when the water immersion 2 of amount is small, the dregs of a decoction is filtered, obtains filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 50
DEG C, continuous heating stirs 30 minutes, obtains mixed liquid；
3rd step, the NaOH solution that 70% is added in the mixed liquid obtained to second step adjust PH to 7.1, then 110 DEG C,
It sterilizes 15 minutes under the conditions of 110kPa, obtains bacillus licheniformis culture medium.
Embodiment 2
Formula：Corn pulp 9g, peptone 8g, sugarcane 9g, steamed beef soup 30g, roundpod jute seed 9g, rhizoma cyperi 10g, eleusine indica 9g, Huang
Over sixty years of age 9g, Chinese lizardtail rhizome 10g, ammonium chloride 0.3g, calcium bicarbonate 0.2g, phosphoric acid hydrogen zinc 0.3g and niter cake 0.2g.
Preparation process：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 3 times of its total amount
When progress boiling 1 is small after when the water immersion 4 of amount is small, the dregs of a decoction are filtered, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 80
DEG C, when continuous heating stirring 1 is small, obtain mixed liquid；
3rd step, the NaOH solution that 80% is added in the mixed liquid obtained to second step adjust PH to 7.3, then 130 DEG C,
It sterilizes 18 minutes under the conditions of 120kPa, obtains bacillus licheniformis culture medium.
Embodiment 3
Formula：Corn pulp 6g, peptone 4g, sugarcane 6g, steamed beef soup 25g, roundpod jute seed 5g, rhizoma cyperi 6g, eleusine indica 6g, astragalus
5g, Chinese lizardtail rhizome 6g, ammonium chloride 0.15g, calcium bicarbonate 0.1g, phosphoric acid hydrogen zinc 0.1g and niter cake 0.15g.
Preparation process：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 2 times of its total amount
When progress boiling 1 is small after when the water immersion 3 of amount is small, the dregs of a decoction are filtered, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 60
DEG C, when continuous heating stirring 1 is small, obtain mixed liquid；
3rd step, the NaOH solution that 80% is added in the mixed liquid obtained to second step adjust PH to 7.2, then 120 DEG C,
It sterilizes 16 minutes under the conditions of 120kPa, obtains bacillus licheniformis culture medium.
Embodiment 4
Formula:Corn pulp 8g, peptone 7g, sugarcane 9g, steamed beef soup 30g, roundpod jute seed 7g, rhizoma cyperi 8g, eleusine indica 9g, Huang
Over sixty years of age 7g, Chinese lizardtail rhizome 8g, ammonium chloride 0.2g, calcium bicarbonate 0.15g, phosphoric acid hydrogen zinc 0.2g and niter cake 0.2g.
Preparation process：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 3 times of its total amount
When progress boiling 1 is small after when the water immersion 4 of amount is small, the dregs of a decoction are filtered, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 80
DEG C, when continuous heating stirring 1 is small, obtain mixed liquid；
3rd step, the NaOH solution that 80% is added in the mixed liquid obtained to second step adjust PH to 7.3, then 130 DEG C,
It sterilizes 18 minutes under the conditions of 120kPa, obtains bacillus licheniformis culture medium.
Embodiment 5
Formula：Corn pulp 8g, peptone 4g, sugarcane 8g, steamed beef soup 24g, roundpod jute seed 8g, rhizoma cyperi 8g, eleusine indica 6g, astragalus
4g, Chinese lizardtail rhizome 5g, ammonium chloride 0.15g, calcium bicarbonate 0.1g, phosphoric acid hydrogen zinc 0.2g and niter cake 0.15g.
Preparation process：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 3 times of its total amount
When progress boiling 1 is small after when the water immersion 4 of amount is small, the dregs of a decoction are filtered, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 80
DEG C, when continuous heating stirring 1 is small, obtain mixed liquid；
3rd step, the NaOH solution that 80% is added in the mixed liquid obtained to second step adjust PH to 7.3, then 130 DEG C,
It sterilizes 18 minutes under the conditions of 120kPa, obtains bacillus licheniformis culture medium.
Embodiment 6
Formula：Corn pulp 7g, peptone 5g, sugarcane 7g, steamed beef soup 27g, roundpod jute seed 6g, rhizoma cyperi 7g, eleusine indica 7g, astragalus
6g, Chinese lizardtail rhizome 7g, ammonium chloride 0.18g, calcium bicarbonate 0.13g, phosphoric acid hydrogen zinc 0.15g and niter cake 0.18g.
Preparation process：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 2 times of its total amount
When progress boiling 1 is small after when the water immersion 3 of amount is small, the dregs of a decoction are filtered, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 60
DEG C, when continuous heating stirring 1 is small, obtain mixed liquid；
3rd step, the NaOH solution that 80% is added in the mixed liquid obtained to second step adjust PH to 7.3, then 130 DEG C,
It sterilizes 16 minutes under the conditions of 120kPa, obtains bacillus licheniformis culture medium.
Contrast test object
The culture medium and traditional bacillus licheniformis culture medium is selected to carry out strain that 1-6 of the embodiment of the present invention is obtained
Cultivate comparison.
Contrast groups medium component：0.4 part of beef extract, 1 part of peptone, 0.6 part of sodium chloride, 1.2 parts of agar powder.
Strain：Bacillus licheniformis Bacillus licheniformis;It is real by Shandong Agricultural University's Life Science College
Room offer is provided.
Condition of culture：38 DEG C of temperature control shaking tables, 265r/min, 48 it is small when.
Production gemma rate contrast table is shown in Table 1
| Group | Temperature | Rotating speed | Time | Produce gemma rate |
| Embodiment 1 | 38℃ | 265r/min | 48 it is small when | 97.4% |
| Embodiment 2 | 38℃ | 265r/min | 48 it is small when | 97.5% |
| Embodiment 3 | 38℃ | 265r/min | 48 it is small when | 97.4% |
| Embodiment 4 | 38℃ | 265r/min | 48 it is small when | 97.5% |
| Embodiment 5 | 38℃ | 265r/min | 48 it is small when | 98.1% |
| Embodiment 6 | 38℃ | 265r/min | 48 it is small when | 98% |
| Contrast groups | 38℃ | 265r/min | 48 it is small when | 68% |
As can be seen from Table 1, the bacillus licheniformis production gemma rate of medium culture of the invention will be far above traditionally
Clothing bacillus culture medium.
Bacteriostatic activity compares
Experimental subjects carries out bacteriostasis property comparison to seven groups of obtained lichem bacillus strains as a comparison above.
Opportunistic pathogen mycelia：Salmonella, enterococcus, Escherichia coli.
Mycelial growth inhibits method：The opportunistic pathogen mycelia of 4 DEG C of preservations is transferred on PDA tablets and is activated, at 28 DEG C
Culture 4-6d, size one is broken into the aseptic card punch of a diameter of 5mm in pathogen thalline marginal position respectively in insulating box
The bacterium dish of cause is respectively connected to transfer needle picking bacterium dish in PDA tablets, is being accessed at bacterium dish 3mm using point connection
Bacillus licheniformis is placed in 28 DEG C of insulating boxs after cultivating 3-5d and measures six groups of embodiments and control group lichens bud respectively
The inhibition zone radius of spore bacillus.
Comparing result is as follows：
Table 2 is the inhibition zone radius of each group（cm）Comparison
| Group | Salmonella | Enterococcus | Escherichia coli |
| Embodiment 1 | 1.83 | 1.38 | 1.12 |
| Embodiment 2 | 1.84 | 1.36 | 1.14 |
| Embodiment 3 | 1.86 | 1.37 | 1.13 |
| Embodiment 4 | 1.89 | 1.37 | 1.15 |
| Embodiment 5 | 1.88 | 1.41 | 1.17 |
| Embodiment 6 | 1.89 | 1.42 | 1.18 |
| Control group | 0.96 | 0.75 | 0.65 |
As can be seen from Table 2, the bacillus licheniformis of six embodiments is compared with control group bacillus licheniformis, antibacterial work
It is with obvious effects to be higher than control group bacterial strain.
To sum up, culture medium of the invention is applied to the culture of bacillus licheniformis, and gemma yield is high, at low cost, and
The bacillus licheniformis arrived has higher bacteriostasis property, and stability is good.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all the present invention spirit and
Within principle, any modifications, equivalent replacements and improvements are made should all be included in the protection scope of the present invention.
Claims (4)
1. a kind of bacillus licheniformis culture medium, which is characterized in that be made by the raw material of following parts by weight：4-9 parts of corn pulp,
3-8 parts of peptone, 5-9 parts of sugarcane, 20-30 parts of steamed beef soup, 3-9 parts of roundpod jute seed, 4-10 parts of rhizoma cyperi, 5-9 parts of eleusine indica, astragalus 3-
9 parts, 4-10 parts of Chinese lizardtail rhizome, 0.1-0.3 parts of ammonium chloride, 0.05-0.2 parts of calcium bicarbonate, 0.05-0.3 parts of phosphoric acid hydrogen zinc and sulphur
Sour 0.1-0.2 parts of hydrogen sodium；
The preparation method of the bacillus licheniformis culture medium is：
The first step takes sugarcane, roundpod jute seed, rhizoma cyperi, eleusine indica, astragalus and Chinese lizardtail rhizome according to proportioning, adds in 1-3 times of its total amount and measures
Water impregnate 2-4 it is small when after carry out boiling 30 minutes -1 it is small when, filter the dregs of a decoction, obtain filtrate；
Surplus stock according to proportioning is mixed, and adds in the filtrate that the first step obtains by second step, is stirred and is heated to 50-80 DEG C,
Continuous heating stir 30 minutes -1 it is small when, obtain mixed liquid；
3rd step, the NaOH solution that 70-80% is added in the mixed liquid obtained to second step adjusts PH to 7.1-7.3, then in 110-
130 DEG C, sterilize 15-18 minutes under the conditions of 110-120kPa, obtain bacillus licheniformis culture medium.
2. bacillus licheniformis culture medium according to claim 1, which is characterized in that by the raw material system of following parts by weight
：6-8 parts of corn pulp, 4-7 parts of peptone, 6-9 parts of sugarcane, 25-30 parts of steamed beef soup, 5-7 parts of roundpod jute seed, 6-8 parts of rhizoma cyperi, Cowhells
6-9 parts careless, 5-7 parts of astragalus, 6-8 parts of Chinese lizardtail rhizome, 0.15-0.2 parts of ammonium chloride, 0.1-0.15 parts of calcium bicarbonate, phosphoric acid hydrogen zinc
0.1-0.2 parts and 0.15-0.2 parts of niter cake.
3. bacillus licheniformis culture medium according to claim 1, which is characterized in that by the raw material system of following parts by weight
：8 parts of corn pulp, 4 parts of peptone, 8 parts of sugarcane, 24 parts of steamed beef soup, 8 parts of roundpod jute seed, 8 parts of rhizoma cyperi, 6 parts of eleusine indica, astragalus 4
Part, 5 parts of Chinese lizardtail rhizome, 0.15 part of ammonium chloride, 0.1 part of calcium bicarbonate, 0.2 part of phosphoric acid hydrogen zinc and 0.15 part of niter cake.
4. bacillus licheniformis culture medium according to claim 1, which is characterized in that by the raw material system of following parts by weight
：7 parts of corn pulp, 5 parts of peptone, 7 parts of sugarcane, 27 parts of steamed beef soup, 6 parts of roundpod jute seed, 7 parts of rhizoma cyperi, 7 parts of eleusine indica, astragalus 6
Part, 7 parts of Chinese lizardtail rhizome, 0.18 part of ammonium chloride, 0.13 part of calcium bicarbonate, 0.15 part of phosphoric acid hydrogen zinc and 0.18 part of niter cake.
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