CN104288187B - A kind of preparation method of Ganoderma Lucidum solid fermentation extract - Google Patents
A kind of preparation method of Ganoderma Lucidum solid fermentation extract Download PDFInfo
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Abstract
The present invention provides a kind of preparation method of Ganoderma Lucidum solid fermentation extract, belong to natural drug preparation field, this method is culture matrix using extracting the pseudo-ginseng residue after arasaponin, squeezing bagasse, rice bran after sugar etc., new two-way solid fermentation is carried out to lucidum strain, this method is compared with traditional artificial cultivation and deep liquid fermentation method, it the advantage is that with short production cycle, tunning ganoderma polyoses content >=50%, technique is simple, beneficial to industrialization, the tunning can as functional health-care food raw material.
Description
Technical field
The invention belongs to natural drug preparation field, and in particular to a kind of preparation side of Ganoderma Lucidum solid fermentation extract
Method.
Background technology
Ganoderma lucidum (Ganoderma lucidum), belongs to the drying fructification of basidiomycetes Polyporaceae red sesame or purple sesame, is
A kind of macro fungi, there is " celestial grass ", the title of " seocho ".There is dual regulation to human body, disease is controlled, be related to heart and brain blood
Each system such as pipe, digestion, nerve, endocrine, breathing, motion, ganoderma lucidum has inhibitory action to nervous system, had to the circulatory system
Step-down and the effect for strengthening cardiac contractile force, have phlegm-dispelling functions to respiratory system.In addition, also protect liver, improve immunologic function, resist
Bacterium etc. acts on, especially very notable to tumour, hepatic disease, insomnia and the preventive and therapeutic effect of aging.
GL-B is the primary bioactive components of ganoderma lucidum, and GL-B is present in natural glossy ganoderma fructification and mycelium
In, GL-B has antitumor, antioxidant radical, anti-aging, improves the bioactivity such as immunity, promoting blood circulation and removing blood stasis.Due to day
The right fructification production cycle is long, and climate influence is big, and yield and quality are unstable, and fructification degree of lignification is high, and polysaccharide is carried
Take utilization rate relatively low.Mycelium is produced using biofermentation technique, the cycle is short, low cost, yield are big, and with before industrialized production
Scape.
The common fermentation of medicinal plant and medicinal fungi, can the comprehensive advantage of the two, it may be found that new natural drug and new
Pharmacological active substance, exploitation co-fermentation technology be Chinese medicine and natural drug innovation direction.
The content of the invention
The invention aims to solve the deficiencies in the prior art, there is provided a kind of system of Ganoderma Lucidum solid fermentation extract
Preparation Method, this method is in preparation process, and culture subenvironment is equipped with the pseudo-ginseng residue extracted after arasaponin, squeezed after sugar
The low pressure polyethylene bags of the culture matrixes such as bagasse.This method is with short production cycle, active constituent content is high, management is easy, is not required to
Want special chemical reagent.
The technical solution adopted by the present invention is as follows：
A kind of preparation method of Ganoderma Lucidum solid fermentation extract, comprises the following steps：
Step（1）, lucidum strain preparation：Ganoderma lucidum slant strains are activated in first class inoculum nutrient solution, 4-5 is grown
My god, obtain first class inoculum；First class inoculum is transferred in secondary bacterial culture liquid after separation and cultivated, two grades of bacterium are obtained after 4-5 days
Kind；
Step（2）, solid medium preparation：1.8-2.2 in mass ratio:0.9-1.1:0.9-1.1:6 weigh pseudo-ginseng residue,
Bagasse, rice bran and purified water, after fully mixing thoroughly, are loaded into culture bag, and every bag of weight is 1.4-1.7kg, and sterilize standby
With obtaining solid medium；
Step（3）, inoculation fermentation：By step（1）Obtained second class inoculum is forwarded to step（2）Obtained solid medium
Interior, inoculum concentration is 10-30%, and both sides are communicated with oxygen supply with the external world when inoculation finishes bundle bag；Solid medium is placed in temperature after inoculation
Light culture is carried out under 25-30 DEG C of degree, humidity 60-75% culture environment；
Step（4）, culture terminal determination：Treat solid medium to cover with ganoderma lucidum mycelium and can terminate before not long fructification
Culture；
Step（5）, the preparation of co-fermentation extract：Extracted to covering with mycelial solid medium, extract solution warp
Concentrate, dry, obtain common fermentation product, i.e. Ganoderma Lucidum solid fermentation extract, ganoderma lucidum is more in the Ganoderma Lucidum solid fermentation extract
Sugared content >=50%, content of ashes≤10%.
Above-mentioned technical proposal step（1）Described in slant strains be inoculated in what PDA culture medium was obtained for lucidum strain, institute
The PDA culture medium stated is potato agar culture medium, and pH value is natural.
Above-mentioned technical proposal step（1）Described in one-level, secondary bacterial culture liquid is including in parts by weight
Following component：200 parts of potato, 20 parts of glucose, 2 parts of peptone, KH2PO41 part, MgSO40.6 part, 1000 parts of water；PH is certainly
So.
Above-mentioned technical proposal step（2）Described in solid medium pseudo-ginseng residue, the mass ratio of bagasse and rice bran be
2：1：1.
Above-mentioned technical proposal, it is further preferred that described pseudo-ginseng residue is residual to extract the pseudo-ginseng after arasaponin
Slag, described bagasse is the bagasse after squeezing sugar.
Above-mentioned technical proposal, step（2）Described in culture bag be low pressure polyethylene bags.
Above-mentioned technical proposal step（3）Described in bundle bag concrete operations be by the two ends of solid medium with PTFE without
Bacterium sealing ventilated membrane sealing.
Above-mentioned technical proposal step（3）Described in the concrete operations of inoculation be：The standby solid medium and two of sterilizing
Level strain liquid is inoculated with sterile working, makes a call to three holes in solid medium top uniform vertical with long handle tweezers, often
Individual hole is inoculated with 15-20ml strain liquid, then covers with culture medium the hole of inoculation.
Step（4）Described in incubation time be 15-25 days.
Above-mentioned technical proposal step（5）Described in extracting method for mycelial culture medium quality multiple will be covered with
1.5-2.5 times is measured, and concentration is that 85% alcohol reflux is extracted 2-3 times, and filtering, merging filtrate obtains ganoderma lucidum total fermentation material ethanol extract,
Be concentrated into it is sticky after, be contained in baking tray in 70 DEG C of air dry oven dry, obtain dried object A；Filter residue quality multiple
1.5-2.5 times of purified water circumfluence distillation for measuring 2-3 times, filtering, merging filtrate obtains ganoderma lucidum total fermentation material aqueous extract, concentrates,
Spray drying, obtains dried object B；Dried object A and dried object B is merged, Ganoderma Lucidum solid fermentation extract is produced.
Compared with prior art, its advantage is the present invention：
1st, the inventive method is with short production cycle compared with traditional artificial cultivation and deep liquid fermentation method, yield and
Steady quality, beneficial to industrialization, Ganoderma Lucidum solid fermentation extract produced by the invention can as functional health-care food original
Material；The present invention by made full use of in the step of preparing Ganoderma Lucidum solid fermentation extract extract saponin(e after pseudo-ginseng residue and
The bagasse after sugar is squeezed, the comprehensive utilization ratio of waste residue is improved, reduces cost；Promote the life of Ganoderma Lucidum to a certain extent simultaneously
Long and increase fermentate polyoses content.
2nd, the present invention does not need special chemical reagent, and Extraction solvent is mainly water and ethanol, and safety non-toxic is conducive to production
The safety in production of product and the protection to environment.
3rd, the present invention is in the step of preparing Ganoderma Lucidum solid fermentation extract, and culture overall situation is under specific humiture
Light culture, management is easy.
Embodiment
With reference to embodiment, the present invention is described in further detail.
Medicine used in the present invention, reagent and instrument are ordinary commercial products.
Embodiment 1
A kind of preparation method of Ganoderma Lucidum solid fermentation extract, comprises the following steps：
Step（1）, lucidum strain preparation：Lucidum strain is inoculated in ganoderma lucidum slant strains that PDA culture medium obtains in one
Activated in level bacteria culture fluid, grow 4 days, obtain first class inoculum；First class inoculum is transferred to second class inoculum culture after separation
Cultivated in liquid, second class inoculum is obtained after 4 days；Wherein, described one-level, secondary bacterial culture liquid are including according to parts by weight
The following component of number meter：200 parts of potato, 20 parts of glucose, 2 parts of peptone, KH2PO41 part, MgSO40.6 part, 1000 parts of water；
PH is natural；
Step（2）, solid medium preparation：In mass ratio 1.8:0.9:1.1:6 weigh pseudo-ginseng residue, bagasse, rice bran
And purified water, after fully mixing thoroughly, it is loaded into low pressure polyethylene bags, every bag of weight is 1.4kg, standby after sterilizing, consolidate
Body culture medium；
Step（3）, inoculation fermentation：By step（1）Obtained second class inoculum is forwarded to step（2）Obtained solid medium
Interior, inoculum concentration is 10%, and inoculation, which is finished, seals at the two ends of solid medium with the sterile sealing ventilated membranes of PTFE；Will be solid after inoculation
Body culture medium is placed under 25 DEG C of temperature, the culture environment of humidity 60% and carries out light culture；The concrete operations of described inoculation are：Go out
The standby solid medium of bacterium and secondary bacteria liquid are inoculated with sterile working, with long handle tweezers on solid medium top
End uniform vertical makes a call to three holes, each hole inoculation 15ml strain liquid, then covers with culture medium the hole of inoculation；
Step（4）, culture terminal determination：Treat solid medium to cover with ganoderma lucidum mycelium and can terminate before not long fructification
Culture, described incubation time is 15 days；
Step（5）, the preparation of co-fermentation extract：Mycelial 1.5 times of amounts of culture medium quality multiple, concentration will be covered with
Extracted 3 times for 85% alcohol reflux, filtering, merging filtrate obtains ganoderma lucidum total fermentation material ethanol extract, be concentrated into it is sticky after, be contained in
Dried in baking tray in 70 DEG C of air dry oven, obtain dried object A；Filter residue is returned with the purifying hydro-thermal of 1.5 times of amounts of quality multiple
Stream is extracted 3 times, and filtering, merging filtrate obtains ganoderma lucidum total fermentation material aqueous extract, is concentrated, and spray drying obtains dried object B；Will be dry
Dry thing A and dried object B merge, and produce Ganoderma Lucidum solid fermentation extract.
Wherein, described pseudo-ginseng residue is extracts the pseudo-ginseng residue after arasaponin, and described bagasse is to squeeze after sugar
Bagasse.
The present embodiment co-cultures 15 bags of solid mediums, obtains Ganoderma Lucidum solid fermentation extract 1250g, polyoses content is
52.01%, content of ashes is 6.00%.
Embodiment 2
A kind of preparation method of Ganoderma Lucidum solid fermentation extract, comprises the following steps：
Step（1）, lucidum strain preparation：Lucidum strain is inoculated in ganoderma lucidum slant strains that PDA culture medium obtains in one
Activated in level bacteria culture fluid, grow 5 days, obtain first class inoculum；First class inoculum is transferred to second class inoculum culture after separation
Cultivated in liquid, second class inoculum is obtained after 5 days；Wherein, described one-level, secondary bacterial culture liquid are including according to parts by weight
The following component of number meter：200 parts of potato, 20 parts of glucose, 2 parts of peptone, KH2PO41 part, MgSO40.6 part, 1000 parts of water；
PH is natural；
Step（2）, solid medium preparation：In mass ratio 2.2: 1.1:0.9:6 weigh pseudo-ginseng residue, bagasse, rice bran
And purified water, after fully mixing thoroughly, it is loaded into low pressure polyethylene bags, every bag of weight is 1.7kg, standby after sterilizing, consolidate
Body culture medium；
Step（3）, inoculation fermentation：By step（1）Obtained second class inoculum is forwarded to step（2）Obtained solid medium
Interior, inoculum concentration is 30%, and inoculation, which is finished, seals at the two ends of solid medium with the sterile sealing ventilated membranes of PTFE；Will be solid after inoculation
Body culture medium is placed under 30 DEG C of temperature, the culture environment of humidity 75% and carries out light culture；The concrete operations of described inoculation are：Go out
The standby solid medium of bacterium and secondary bacteria liquid are inoculated with sterile working, with long handle tweezers on solid medium top
End uniform vertical makes a call to three holes, each hole inoculation 20ml strain liquid, then covers with culture medium the hole of inoculation；
Step（4）, culture terminal determination：Treat solid medium to cover with ganoderma lucidum mycelium and can terminate before not long fructification
Culture, described incubation time is 25 days；
Step（5）, the preparation of co-fermentation extract：Mycelial 2 times of amounts of culture medium quality multiple will be covered with, concentration is
85% alcohol reflux is extracted 2 times, filtering, and merging filtrate obtains ganoderma lucidum total fermentation material ethanol extract, be concentrated into it is sticky after, be contained in roasting
Dried in disk in 70 DEG C of air dry oven, obtain dried object A；The purifying hydro-thermal backflow of 2 times of amounts of filter residue quality multiple is carried
Take 2 times, filter, merging filtrate obtains ganoderma lucidum total fermentation material aqueous extract, concentrate, spray drying obtains dried object B；By dried object A
Merge with dried object B, produce Ganoderma Lucidum solid fermentation extract.
Wherein, described pseudo-ginseng residue is extracts the pseudo-ginseng residue after arasaponin, and described bagasse is to squeeze after sugar
Bagasse.
The present embodiment co-cultures 12 bags of solid mediums, obtains Ganoderma Lucidum solid fermentation extract 1050g, polyoses content is
57.11%, content of ashes is 5.70%.
Embodiment 3
A kind of preparation method of Ganoderma Lucidum solid fermentation extract, comprises the following steps：
Step（1）, lucidum strain preparation：Lucidum strain is inoculated in ganoderma lucidum slant strains that PDA culture medium obtains in one
Activated in level bacteria culture fluid, grow 4.5 days, obtain first class inoculum；First class inoculum is transferred to second class inoculum training after separation
Cultivated in nutrient solution, second class inoculum is obtained after 4.8 days；Wherein, described one-level, secondary bacterial culture liquid are including according to weight
Measure the following component of number meter：200 parts of potato, 20 parts of glucose, 2 parts of peptone, KH2PO41 part, MgSO40.6 part, water
1000 parts；PH is natural；
Step（2）, solid medium preparation：In mass ratio 2:1:1:6 weigh pseudo-ginseng residue, bagasse, rice bran and purifying
Water, after fully mixing thoroughly, is loaded into low pressure polyethylene bags, and every bag of weight is 1.6kg, standby after sterilizing, obtains solid culture
Base；
Step（3）, inoculation fermentation：By step（1）Obtained second class inoculum is forwarded to step（2）Obtained solid medium
Interior, inoculum concentration is 20%, and inoculation, which is finished, seals at the two ends of solid medium with the sterile sealing ventilated membranes of PTFE；Will be solid after inoculation
Body culture medium is placed under 27 DEG C of temperature, the culture environment of humidity 68% and carries out light culture；The concrete operations of described inoculation are：Go out
The standby solid medium of bacterium and secondary bacteria liquid are inoculated with sterile working, with long handle tweezers on solid medium top
End uniform vertical makes a call to three holes, each hole inoculation 18ml strain liquid, then covers with culture medium the hole of inoculation；
Step（4）, culture terminal determination：Treat solid medium to cover with ganoderma lucidum mycelium and can terminate before not long fructification
Culture, described incubation time is 20 days；
Step（5）, the preparation of co-fermentation extract：Mycelial 2.5 times of amounts of culture medium quality multiple, concentration will be covered with
Extracted 2 times for 85% alcohol reflux, filtering, merging filtrate obtains ganoderma lucidum total fermentation material ethanol extract, be concentrated into it is sticky after, be contained in
Dried in baking tray in 70 DEG C of air dry oven, obtain dried object A；Filter residue is returned with the purifying hydro-thermal of 2.5 times of amounts of quality multiple
Stream is extracted 2 times, and filtering, merging filtrate obtains ganoderma lucidum total fermentation material aqueous extract, is concentrated, and spray drying obtains dried object B；Will be dry
Dry thing A and dried object B merge, and produce Ganoderma Lucidum solid fermentation extract.
Wherein, described pseudo-ginseng residue is extracts the pseudo-ginseng residue after arasaponin, and described bagasse is to squeeze after sugar
Bagasse.
The present embodiment co-cultures 15 bags of solid mediums, obtains Ganoderma Lucidum solid fermentation extract 1200g, polyoses content is
54.60%, content of ashes is 5.60%.
Table 1 is the ratio of Ganoderma Lucidum solid fermentation extract polyoses content and other patented methods prepared by the inventive method
Compared with specific as follows：
Table 1
Claims (10)
1. a kind of preparation method of Ganoderma Lucidum solid fermentation extract, it is characterised in that comprise the following steps：
Step（1）, lucidum strain preparation：Ganoderma lucidum slant strains are activated in first class inoculum nutrient solution, grown 4-5 days,
Obtain first class inoculum；First class inoculum is transferred in secondary bacterial culture liquid after separation and cultivated, second class inoculum is obtained after 4-5 days；
Step（2）, solid medium preparation：1.8-2.2 in mass ratio:0.9-1.1:0.9-1.1:6 weigh pseudo-ginseng residue, sugarcane
Slag, rice bran and purified water, after fully mixing thoroughly, are loaded into culture bag, and every bag of weight is 1.4-1.7kg, standby after sterilizing, is obtained
To solid medium；
Step（3）, inoculation fermentation：By step（1）Obtained second class inoculum is forwarded to step（2）In obtained solid medium,
Inoculum concentration is 10-30%, and both sides are communicated with oxygen supply with the external world when inoculation finishes bundle bag；Solid medium is placed in temperature after inoculation
25-30 DEG C, light culture is carried out under humidity 60-75% culture environment；
Step（4）, culture terminal determination：Treat solid medium to cover with ganoderma lucidum mycelium and training can be terminated before not long fructification
Support；
Step（5）, the preparation of co-fermentation extract：Extracted to covering with mycelial solid medium, extract solution is concentrated,
Dry, obtain common fermentation product, i.e. Ganoderma Lucidum solid fermentation extract, ganoderma polyoses content in the Ganoderma Lucidum solid fermentation extract
>=50%, content of ashes≤10%.
2. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（1）Middle institute
The slant strains stated are inoculated in what PDA culture medium was obtained for lucidum strain.
3. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（1）Middle institute
The one-level stated, secondary bacterial culture liquid are including following component in parts by weight：200 parts of potato, glucose 20
Part, 2 parts of peptone, KH2PO41 part, MgSO40.6 part, 1000 parts of water；PH is natural.
4. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（2）Middle institute
The mass ratio of pseudo-ginseng residue, bagasse and rice bran is 2 in the solid medium stated：1：1.
5. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 4, it is characterised in that described pseudo-ginseng
Residue is extracts the pseudo-ginseng residue after arasaponin, and described bagasse is the bagasse after squeezing sugar.
6. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（2）Middle institute
The culture bag stated is low pressure polyethylene bags.
7. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（3）Middle institute
The concrete operations for the bundle bag stated are to seal at the two ends of solid medium with the sterile sealing ventilated membranes of PTFE.
8. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（3）Middle institute
The concrete operations for the inoculation stated are：The standby solid medium of sterilizing and secondary bacteria liquid are inoculated with sterile working room,
Three holes are made a call in solid medium top uniform vertical, each hole is inoculated with 15-20ml strain liquid with long handle tweezers, then with cultivating
Base lid lives inoculation hole.
9. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（4）Middle institute
The incubation time stated is 15-25 days.
10. the preparation method of Ganoderma Lucidum solid fermentation extract according to claim 1, it is characterised in that step（5）In
Described extracting method is measured for 1.5-2.5 times for that will cover with mycelial culture medium with quality multiple, and concentration carries for 85% alcohol reflux
Take 2-3 times, filter, merging filtrate obtains ganoderma lucidum total fermentation material ethanol extract, be concentrated into it is sticky after, in 70 DEG C of air dry oven
Middle drying, obtains dried object A；The purified water circumfluence distillation measured with 1.5-2.5 times of quality multiple of filter residue 2-3 times, filters, merges
Filtrate obtains ganoderma lucidum total fermentation material aqueous extract, concentrates, and spray drying obtains dried object B；Dried object A and dried object B is merged,
Produce Ganoderma Lucidum solid fermentation extract.
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| Title |
|---|
| 中药渣用于培养灵芝菌及功能成分分析;陈合等;《农业工程学报》;20061231;第22卷;第167-170页，尤其是第167页第1.2节，第168页第1.3.2节、第2.2.2节表4、第2.3.2节，第169页第2.3.6节及第169页右栏第2段第1-2行 * |
| 几种食用真菌对甘蔗渣利用的研究;吴萍等;《安徽技术师范学院学报》;20041231;第18卷(第2期);第9-11页 * |
| 利用中药渣培养灵芝菌及生物活性成分的研究;陈合等;《食品工业科技》;20061231;第27卷(第10期);第58-60,63页 * |
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