CN106906205A - The method for promoting grifola frondosus strain liquid state fermentation using magnetic field - Google Patents
The method for promoting grifola frondosus strain liquid state fermentation using magnetic field Download PDFInfo
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Abstract
The method for promoting grifola frondosus strain liquid state fermentation using magnetic field is disclosed in the present invention, belongs to food processing technology field.Methods described step is as follows：（1）Grifola frondosus strain after activation is inoculated in seed culture medium, through seed culture, seed liquor is obtained；（2）By step（1）Obtained seed liquor is transferred in fermentation medium, periodically adds magnetic field aid in treatment to promote fermentation in its growth cycle, obtains the tunning after the treatment of magnetic field.Mycelial biomass can be made to improve about 12% than normal fermentation product under optimal conditions, the method step is simple, it is easy to operate, it is possible to decrease the production cost of maitake mushroom mycelia.
Description
Technical field
The invention belongs to food processing technology field, it is specifically designed a kind of using magnetic field treatment promotion grifola frondosus liquid state fermentation
Method.
Background technology
Grifola frondosus belong to Basidiomycotina, Hymenomycetes, Aphyllophorales, Polyporaceae, a kind of medicine of tree flower Pseudomonas, food dual-purpose
High-grade edible bacterium, be that WHO and FAO organizes one of the rare economic kind recommended to developing country.Grifola Frondosa sporophore meat
Matter is tender and crisp, tasty mouthfeel, crude protein content 23.96%, total sugar content 48.05%, total amino acid content in its extract powder
12.66mg/100g (essential amino acid accounts for the 41% of total amino acid content), additionally containing taurine 266.83mg/100g and
Abundant mineral element, it is nutritious.Experiment both at home and abroad shows that grifola frondosus have extensive pharmacological activity, such as antitumor, antioxygen
The effects such as change, radioresistance and regulation immunologic function.(traditional Chinese medicines are accurate for the current existing pharmaceutical producing enterprise's production grifola frondosus capsule of China
Word B20020023).The industrialized production of current grifola frondosus still relies primarily on solid state fermentation, and liquid submerged fermentation is compared to solid
State fermentation is obtaining bioactive product such as polysaccharide of mycelium and stabilization etc. more effectively.Therefore, by studying grifola frondosus liquid
State deep drainpipe condition is a kind of novel to develop, and the method for efficiently obtaining maitake mushroom mycelia (and product) turns into what is received much concern
Problem.
In recent years, biomagnetism gos deep into magnetic field and is widely applied to as a new type edge subject with research
Industrial and agricultural production.Gao Mengxiang etc. is by studying the alternating magnetic field under discovery certain condition to brewer's yeast (0.563~0.669A/
M), the growth of the strain such as Hericium erinaceus (0.42~3.04A/m) has facilitation (Gao Mengxiang, Xia Fan, Hu Qiudong alternating magnetic fields
Growth to brewer's yeast promotes effect [J] agricultural mechanical journals, 2007,38 (7):91~93；Gao Mengxiang, Xia Fan, Zhu Peng
Research [J] agricultural mechanical journals that great waves alternating magnetic fields are acted on mycelium growth of Hericium erinaceus, 2009,40 (2):139~141).It is so far
Only, promote frondosa fermentation by using short period, the treatment of more low intensive magnetic field, improve the text of maitake mushroom mycelia yield
There is not been reported for chapter or patent.
The content of the invention
It is an object of the invention to provide a kind of method that utilization magnetic field promotes grifola frondosus strain liquid state fermentation.
Realize that technology of the invention is as follows：
To achieve the above object, the technical scheme taken of the present invention is：Promote grifola frondosus strain liquid state fermentation using magnetic field
Method, carry out as steps described below：
(1) culture medium and maitake mushroom mycelia condition of culture
Grifola frondosa strain (Griflolafrondosa), is seeded in aseptic PDA inclined-planes shaking speed for 180r/min, and 28
DEG C culture 5 days after make nutrient solution by the use of homogenizer in mycelium pellet be uniformly dispersed as seed liquor.Seed culture fluid is pressed into 10% (v/
V) inoculum concentration is seeded to containing 100mL in the 250mL conical flasks without bacteria fermentation culture medium, and conical flask is placed on shaking table
150r/min, 28 DEG C are cultivated and carried out magnetic field aid in treatment.
Wherein PDA culture medium formula：Potato leaches powder 6g/L, glucose 20g/L, MgS040.75g/L, KH2P04
1.5g/L, soy peptone 5g/L, agar 20g/L, pH nature) upper 28 DEG C of trainings culture, 5 pieces big will be cut off after 5 days from inclined-plane
The about mycelia of 5 × 5mm, and be inoculated in the conical flask of 250mL, wherein containing 100mL aseptic seed culture mediums.
Wherein PDA liquid medium formula：Potato leaches powder 6g/L, glucose 20g/L, MgS040.75g/L,
KH2P041.5g/L, soy peptone 5g/L, pH nature.
Wherein fermentative medium formula：Potato leaches powder 6g/L, glucose 20g/L, MgS040.75g/L, KH2P04
1.5g/L, soy peptone 5g/L, pH nature.
(2) magnetic field assisted fermentation
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.Different time points (1-23h) after inoculation proceed by magnetic field treatment, are then carried out again in same time point daily
Magnetic field is processed, coprocessing 5 days.Handling duration is 1-7h, and magnetic induction intensity is 15-55Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.
The present invention promotes maitake mushroom mycelia fermentation using magnetic field technique, and mycelial biomass is determined.By
Magnetic field aid in treatment can significantly improve maitake mushroom mycelia biomass, and process is simple, efficient.
Brief description of the drawings
Fig. 1 is magnetic field means figure used in the present invention, wherein 1 is the shaking table hole of coiling；2 is coil (6 shaking table holes
In the same size, institute's coiling parameter is consistent)；3 is rotational speed regulation button；4 is rotating speed display screen；5 is shaking table；6 is TDGC2-
The single-phase contact voltage regulators of 3k3000VA；7 is 220V AC powers.
Specific embodiment
Magnetic field means used in the present invention are as shown in the figure (Fig. 1).220V AC powers are by the series connection shaking table of pressure regulator 6 hole
Coil 2 on 1, the size for changing voltage in coil by pressure regulator 6 (can come the magnetic induction intensity in the generation of regulating winding 2 magnetic field
Adjustable magnetic induction scope is 15-55Gs)；The rotating speed of shaking table 5 is adjusted by rotational speed regulation button 3, and concrete numerical value is shown in that rotating speed shows
Screen 4；Sample is placed in when magnetic field is processed the center of shaking table hole 1 of coiling.Cultivation temperature passes through airconditioning control.
Grifola frondosa strain (Griflolafrondosa) of the present invention has been applied and granted mandate Chinese invention patent, patent
Number：ZL201110150888.3, title：The bacterial strain of grifolan is produced for ferment rice bran and wheat bran extract solution.
Used as evaluation index, its determination step is mycelium dry weight in the present invention：By the mycelium centrifugation obtained by fermentation
(5000r/min, 10min), the nutrient solution that surface is sticked is removed with distilled water flushing, is put into air dry oven, 50 DEG C of bars
Dried to permanent quality under part, electronic balance weighing obtains final product mycelia dry mass.
The computing formula of mycelium dry weight increase rate is as follows：
Wherein L is mycelium dry weight increase rate, and m is after each magnetic field assisted fermentation condition is processed
Grifola frondosus mycelia dry mass, n is control group grifola frondosus mycelia dry mass.
Reference examples
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.Grifola frondosus hypha fermentation overall process is processed without magnetic field.After fermentation ends, mycelia is by being centrifuged (5000 turns, 10 minutes) receipts
Collection, and after being washed twice with distilled water, dried in 50 DEG C of baking ovens.After constant weight, its mycelial biomass is determined.
Embodiment 1
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 1h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 2
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 2h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 3
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 4
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 4h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 5
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 5h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 6
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 6h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 7
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 7h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 8
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 15Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 9
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 25Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 10
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 45Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 11
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.1h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 55Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 12
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.8h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 13
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.13h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 14
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.18h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
Embodiment 15
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is
5 days.23h proceeds by magnetic field treatment after inoculation, then carries out magnetic field treatment again in same time point daily, until fermentation
Terminate.Magnetic field treatment conditions are：Daily handling duration is 3h, magnetic induction intensity is 35Gs.Daily after magnetic treatment terminates, rapidly
Sample is back on shaking table and is persistently cultivated.The same reference examples of biomass estimation after fermentation ends.Maitake mushroom mycelia biomass is shown in
Table 1.
The different embodiment maitake mushroom mycelia biomass of table 1
As can be seen from the above table, maitake mushroom mycelia fermentation is by after the aid in treatment of magnetic field, mycelial biomass has difference
Degree is improved.Especially fermented and cultured magnetic field aid in treatment condition is：The magnetic treatment time started is 1h, daily magnetic treatment after inoculation
Shi Changwei 3h, magnetic induction intensity are 35Gs, and maitake mushroom mycelia biomass is improved compared to control conspicuousness after fermentation ends
11.43%.
Claims (4)
1. the method for promoting grifola frondosus strain liquid state fermentation using magnetic field, it is characterised in that carry out as steps described below：
（1）Culture medium and maitake mushroom mycelia condition of culture：
Grifola frondosa strain（Griflolafrondosa）, aseptic PDA inclined-planes shaking speed is seeded in for 180r/min, 28 DEG C of trainings
Mycelium pellet is uniformly dispersed as seed liquor in making nutrient solution by the use of homogenizer after supporting 5 days；Seed culture fluid is pressed 10%（v/v）Connect
The amount of kind is seeded to containing 100mL in the 250mL conical flasks without bacteria fermentation culture medium, and conical flask is placed in 150r/ on shaking table
Min, 28 DEG C are cultivated and carried out magnetic field aid in treatment；
（2）Magnetic field assisted fermentation：
Fermentation is calculated under the conditions of 28 DEG C, rotating speed 150r/min since inoculation time, and total fermentation number of days is 5 days；
1-23h after inoculation proceeds by magnetic field treatment, then carries out magnetic field treatment, coprocessing 5 again in same time point daily
My god；Handling duration is 1-7h, and magnetic induction intensity is 15-55Gs；Daily after magnetic treatment terminates, sample is back to shaking table rapidly
Upper lasting culture, you can reach the purpose for promoting grifola frondosus strain liquid state fermentation.
2. the method that utilization magnetic field according to claim 1 promotes grifola frondosus strain liquid state fermentation, it is characterised in that wherein
PDA culture medium is formulated：Potato leaches powder 6g/L, glucose 20g/L, MgS040.75g/L, KH2P041.5g/L, soybean protein
Peptone 5g/L, agar 20g/L, pH nature）Upper 28 DEG C of trainings culture, will cut off 5 pieces of about bacterium of 5 × 5mm from inclined-plane after 5 days
Silk, and be inoculated in the conical flask of 250mL, wherein containing 100mL aseptic seed culture mediums.
3. the method that utilization magnetic field according to claim 1 promotes grifola frondosus strain liquid state fermentation, it is characterised in that wherein
PDA liquid medium is formulated：Potato leaches powder 6g/L, glucose 20g/L, MgS040.75g/L, KH2P041.5g/L, soybean
Peptone 5g/L, pH nature.
4. the method that utilization magnetic field according to claim 1 promotes grifola frondosus strain liquid state fermentation, it is characterised in that wherein
Fermentative medium formula：Potato leaches powder 6g/L, glucose 20g/L, MgS040.75g/L, KH2P041.5g/L, soybean egg
White peptone 5g/L, pH nature.
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| CN109479622A (en) * | 2018-11-15 | 2019-03-19 | 江苏强农农业技术服务有限公司 | A kind of tea tree mushroom strains industrial production method |
| CN109679846A (en) * | 2019-01-23 | 2019-04-26 | 江苏大学 | A kind of method of rotary magnetic field treating device and its forced fermentation |
| CN114875019A (en) * | 2022-05-24 | 2022-08-09 | 深圳劲创生物技术有限公司 | Method for improving toadstool mycelium biomass by magnetic treatment |
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