CN109762746A - A method of addition lignocellulosic activation induction mushroom liquid bacterial - Google Patents
A method of addition lignocellulosic activation induction mushroom liquid bacterial Download PDFInfo
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- CN109762746A CN109762746A CN201910231452.3A CN201910231452A CN109762746A CN 109762746 A CN109762746 A CN 109762746A CN 201910231452 A CN201910231452 A CN 201910231452A CN 109762746 A CN109762746 A CN 109762746A
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Abstract
The invention discloses a kind of methods of addition lignocellulosic activation induction mushroom liquid bacterial, comprising the following steps: (1) pipettes a small amount of refrigeration solid mushroom strain and be inoculated on the inclined-plane of PDA culture medium and cultivated；(2) by the plate of the mycelium inoculation on PDA culture medium inclined-plane to PDA culture medium, back-off is cultivated；(3) step (2) cultured mushroom mycelium is inoculated in seed culture medium and is cultivated, obtain mushroom liquid bacterial to be induced；(4) mushroom liquid bacterial is placed in activation induced medium and is cultivated, the activation induced medium includes the component of following weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%, KH2PO40.1%, MgSO4·7H2O 0.05%, lignocellulosic 1-3%, remaining is distilled water.The biological activity of mushroom liquid bacterial can be improved by this method, improve spawn activity.
Description
Technical field
The present invention relates to edible mushroom culture technique fields, activate induction mushroom more particularly to a kind of addition lignocellulosic
The method of liquid spawn.
Background technique
Mushroom (scientific name: Lentinus edodes) also known as thick mushroom, dried mushroom, mushroom, are a kind of edible fungus.Belong to Agaricales,
Tricholomataceae, Lentinus.Mushroom grows thickly, Dan Sheng or all living creatures.Nutrition needed for mushroom growth can be incited somebody to action by mycelia to Extracellular enzyme
Lignocellulosic and organic macromolecule material reabsorb after decomposing.Mushroom contains a variety of nutriments of needed by human body, can be improved
Immunity is a kind of good medical product.But the mushroom growth period is long, to mushroom using there are drawbacks.Due to natural ring
The limitation in border can also waste many human and material resources.It can produce ectoenzyme during mushroom growth, utilize born of the same parents caused by detection
Exoenzyme can further measure mushroom growth activity.
Lignocellulosic derives from plant, is the organic renewable resource of content highest in nature.It is natural reproducible
Organic wadding fibrous substance that timber is processed by chemical treatment, Mechanical Method, it is nontoxic, tasteless, pollution-free, "dead".
Wood cellulose mainly includes three kinds of compositions: lignin, cellulose, hemicellulose, can be by lignocellulolytic enzymes system
It is degraded to small molecule compound.All contain lignocellulosic such as in agricultural wastes major part: pine tree, poplar, Chinese scholartree.And wood
Matter cellulose is reusable, cheap and easy to get, has certain economic value to the recycling of lignocellulosic.
Summary of the invention
The object of the present invention is to provide a kind of methods of addition lignocellulosic activation induction mushroom liquid bacterial.It utilizes
Carbon source of this 3 kinds of lignocellulosics of pine tree, poplar, Chinese scholartree as induction mushroom liquid bacterial growth, promotes mushroom strain
Activation and growth, obtain the mushroom product of high-quality.
A method of addition lignocellulosic activation induction mushroom liquid bacterial, comprising the following steps:
(1) it pipettes the solid mushroom strain refrigerated on a small quantity and is inoculated on the inclined-plane of PDA culture medium and cultivated；
(2) by the plate of the mycelium inoculation on PDA culture medium inclined-plane to PDA culture medium, back-off is cultivated；
(3) step (2) cultured mushroom mycelium is inoculated in seed culture medium and is cultivated, obtain perfume (or spice) to be induced
Mushroom liquid strain；
(4) mushroom liquid bacterial is placed in activation induced medium and is cultivated, the activation induced medium includes
The component of following weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%,
KH2PO40.1%, MgSO4·7H2O 0.05%, lignocellulosic 1-3%, remaining is distilled water.
Preferably, the PDA culture medium include following weight component: potato 200g, glucose 20g, agar 20g,
Water 1000mL.
Preferably, the condition of culture of the step (1) is to cultivate 9d at 25 DEG C, allows mycelium to cover with inclined-plane, then in 4
Preservation inclined-plane mycelium under the conditions of DEG C, and periodically strain is passed on.
Preferably, the seed culture medium includes the component of following weight: potato 200g, glucose 20g, peptone
10g, dipotassium hydrogen phosphate 5g, tap water 1000mL.
Preferably, the condition of culture of the step (3) is the static gas wave refrigerator 3d in 26 DEG C of shaking tables, then in revolving speed
8d is cultivated in the shaking table that 150r/min and temperature are 26 DEG C, obtains mushroom liquid bacterial to be induced.
Preferably, the lignocellulosic is Chinese scholartree lignocellulosic, in pine tree lignocellulosic, poplar lignocellulosic
One kind.
Preferably, the condition of culture of the step (4) is to weigh wheat bran, dregs of beans, corn flour, glucose, yeast in proportion
Cream, KH2PO4、MgSO4·7H2O and lignocellulosic, are configured to activation induced medium after being mixed, removing step (3) obtains
The mushroom liquid bacterial arrived cultivates 8d in revolving speed 150r/min and in shaking table that temperature is 26 DEG C.
Preferably, the mushroom liquid bacterial of inoculation accounts for the 5% of activation induced medium total amount.
Compared with prior art, the invention has the following advantages: the present invention utilizes lignocellulosic to mushroom liquid
Strain carries out activation induction, during induction fermentation, can promote mushroom liquid bacterial relevant enzyme biological activity, especially paints
Enzyme, carboxymethylcelluloenzyme enzyme, amylase activity are expected to shorten mushroom liquid to promote the growth and development of mushroom liquid bacterial
Sprout time of the strain in cultivation Bag Material, improves production efficiency；The invention novel and unique, can large-scale promotion use, and take
With low, the cost of agricultural production is greatly reduced.
Specific embodiment
Below with reference to embodiment, the invention will be further described.
Embodiment 1
A method of addition lignocellulosic activation induction mushroom liquid bacterial, comprising the following steps:
(1) the refrigeration solid mushroom strain for pipetting soybean grain size, is inoculated on the inclined-plane of PDA culture medium and is cultivated,
The PDA culture medium includes the component of following weight: potato 200g, glucose 20g, agar 20g, water 1000mL, by bacterium
9d is cultivated at 25 DEG C after kind inoculation, allows mycelium to cover with inclined-plane, then the preservation inclined-plane mycelium under the conditions of 4 DEG C, and periodically
Strain is passed on；
(2) by the plate of the mycelium inoculation on PDA culture medium inclined-plane to PDA culture medium, back-off is cultivated；
(3) step (2) cultured mushroom mycelium is inoculated in seed culture medium and is cultivated, obtain perfume (or spice) to be induced
Mushroom liquid strain, the seed culture medium include the component of following weight: potato 200g, glucose 20g, peptone 10g,
Dipotassium hydrogen phosphate 5g, tap water 1000mL, and condition of culture is the static gas wave refrigerator 3d in 26 DEG C of shaking tables, then in revolving speed 150r/
8d is cultivated in the shaking table that min and temperature are 26 DEG C, obtains mushroom liquid bacterial to be induced；
(4) mushroom liquid bacterial is placed in activation induced medium and is cultivated, the activation induced medium includes
The component of following weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%,
KH2PO40.1%, MgSO4·7H2O 0.05%, Chinese scholartree lignocellulosic 2%, remaining is distilled water.Condition of culture be by than
Example weighs wheat bran, dregs of beans, corn flour, glucose, yeast extract, KH2PO4、MgSO4·7H2O and lignocellulosic, after being mixed
It is configured to activation induced medium, the mushroom liquid bacterial that removing step (3) obtains, in revolving speed 150r/min and temperature is 26 DEG C
Shaking table in the mushroom liquid bacterial cultivating 8d, and be inoculated with account for the 5% of activation induced medium total amount.
Embodiment 2
According to the preparation method of embodiment 1, pine tree lignocellulosic preparation activation induced medium is used instead, to mushroom liquid
Strain carries out induced growth, remaining step is constant, and the specific method is as follows:
(1) the refrigeration solid mushroom strain for pipetting soybean grain size, is inoculated on the inclined-plane of PDA culture medium and is cultivated,
The PDA culture medium includes the component of following weight: potato 200g, glucose 20g, agar 20g, water 1000mL, by bacterium
9d is cultivated at 25 DEG C after kind inoculation, allows mycelium to cover with inclined-plane, then the preservation inclined-plane mycelium under the conditions of 4 DEG C, and periodically
Strain is passed on；
(2) by the plate of the mycelium inoculation on PDA culture medium inclined-plane to PDA culture medium, back-off is cultivated；
(3) step (2) cultured mushroom mycelium is inoculated in seed culture medium and is cultivated, obtain perfume (or spice) to be induced
Mushroom liquid strain, the seed culture medium include the component of following weight: potato 200g, glucose 20g, peptone 10g,
Dipotassium hydrogen phosphate 5g, tap water 1000mL, and condition of culture is the static gas wave refrigerator 3d in 26 DEG C of shaking tables, then in revolving speed 150r/
8d is cultivated in the shaking table that min and temperature are 26 DEG C, obtains mushroom liquid bacterial to be induced；
(4) mushroom liquid bacterial is placed in activation induced medium and is cultivated, the activation induced medium includes
The component of following weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%,
KH2PO40.1%, MgSO4·7H2O 0.05%, pine tree lignocellulosic 3%, remaining is distilled water, condition of culture be by than
Example weighs wheat bran, dregs of beans, corn flour, glucose, yeast extract, KH2PO4、MgSO4·7H2O and lignocellulosic, after being mixed
It is configured to activation induced medium, the mushroom liquid bacterial that removing step (3) obtains, in revolving speed 150r/min and temperature is 26 DEG C
Shaking table in the mushroom liquid bacterial cultivating 8d, and be inoculated with account for the 5% of activation induced medium total amount.
Embodiment 3
According to the preparation method of embodiment 1, poplar lignocellulosic preparation activation induced medium is used instead, to mushroom liquid
Strain carries out induced growth, remaining step is constant, and the specific method is as follows:
(1) the refrigeration solid mushroom strain for pipetting soybean grain size, is inoculated on the inclined-plane of PDA culture medium and is cultivated,
The PDA culture medium includes the component of following weight: potato 200g, glucose 20g, agar 20g, water 1000mL, by bacterium
9d is cultivated at 25 DEG C after kind inoculation, allows mycelium to cover with inclined-plane, then the preservation inclined-plane mycelium under the conditions of 4 DEG C, and periodically
Strain is passed on；
(2) by the plate of the mycelium inoculation on PDA culture medium inclined-plane to PDA culture medium, back-off is cultivated；
(3) step (2) cultured mushroom mycelium is inoculated in seed culture medium and is cultivated, obtain perfume (or spice) to be induced
Mushroom liquid strain, the seed culture medium include the component of following weight: potato 200g, glucose 20g, peptone 10g,
Dipotassium hydrogen phosphate 5g, tap water 1000mL, and condition of culture is the static gas wave refrigerator 3d in 26 DEG C of shaking tables, then in revolving speed 150r/
8d is cultivated in the shaking table that min and temperature are 26 DEG C, obtains mushroom liquid bacterial to be induced；
(4) mushroom liquid bacterial is placed in activation induced medium and is cultivated, the activation induced medium includes
The component of following weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%,
KH2PO40.1%, MgSO4·7H2O 0.05%, poplar lignocellulosic 1%, remaining is distilled water, condition of culture be by than
Example weighs wheat bran, dregs of beans, corn flour, glucose, yeast extract, KH2PO4、MgSO4·7H2O and lignocellulosic, after being mixed
It is configured to activation induced medium, the mushroom liquid bacterial that removing step (3) obtains, in revolving speed 150r/min and temperature is 26 DEG C
Shaking table in the mushroom liquid bacterial cultivating 8d, and be inoculated with account for the 5% of activation induced medium total amount.
Comparative example 1
According to the preparation method of embodiment 1, induction is treated using the activation induced medium for being not added with lignocellulosic
Mushroom liquid bacterial carries out shaken cultivation, remaining step is constant, i.e. step (4) are as follows: is placed in mushroom liquid bacterial and is not added with wood
Cultivated in the activation induced medium of matter cellulose, the activation induced medium for being not added with lignocellulosic include with
The component of lower weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%,
KH2PO40.1%, MgSO4·7H2O 0.05%, remaining be distilled water.Condition of culture is to weigh wheat bran, dregs of beans, jade in proportion
Rice flour, glucose, yeast extract, KH2PO4、MgSO4·7H2O, the activation for being configured to be not added with lignocellulosic after being mixed lure
Lead culture medium, the mushroom liquid bacterial that removing step (3) obtains is cultivated in revolving speed 150r/min and in shaking table that temperature is 26 DEG C
8d, and the mushroom liquid bacterial being inoculated with accounts for the 5% of activation induced medium total amount.
Comparative example 2
According to the preparation method of embodiment 2, by the pine tree wood fibre cellulose content in the activation induced medium of step (4)
It is increased to 4%, is induced using the activation induced medium, remaining inductive condition and method are constant.
When detecting embodiment 1-3 and comparative example 1-2 pine tree lignocellulosic activation induction mushroom liquid bacterial, when different
Between in section in culture medium laccase, carboxymethylcelluloenzyme enzyme, amylase activity, testing result is as shown in table 1.
It is 1 enzyme-activity unit that wherein laccase activity measurement, which is enzyme amount required for aoxidizing 1umol guaiacol according to 1min,
(U)；
CMCase activity measurement is basis in the case where pH4.8 and temperature are 50 DEG C, 1 hour hydrolysis substrate CMC-Na
The required enzyme amount for releasing 1mg reduced sugar is an enzyme activity unit U；
Amylase activity measurement is according at 60 DEG C, pH5.6, and enzyme solution is released with 2% soluble starch solution effects 1h
Enzyme amount needed for putting 0.1mg maltose is 1 enzyme activity unit (U).
Table 1
The mycelium pellet dry weight uniformity of the mushroom strain of induction, and bacterium are activated using the method for 1-3 of the embodiment of the present invention
Pompon dry weight increases by 147% or more compared with comparative example 1, substantially increases spawn activity and mycelia ball weight.Meanwhile from comparison
In the detection data of example 2 it can also be seen that after wood fibre cellulose content is higher than 3%, the activation of strain will be inhibited to lure significantly
It leads.
Used herein a specific example illustrates the principle and implementation of the invention, and above embodiments are said
It is bright to be merely used to help understand method and its core concept of the invention；At the same time, for those skilled in the art, foundation
Thought of the invention, there will be changes in the specific implementation manner and application range.In conclusion the content of the present specification is not
It is interpreted as limitation of the present invention.
Claims (8)
1. a method of addition lignocellulosic activation induction mushroom liquid bacterial, which comprises the following steps:
(1) it pipettes the solid mushroom strain refrigerated on a small quantity and is inoculated on the inclined-plane of PDA culture medium and cultivated；
(2) by the plate of the mycelium inoculation on PDA culture medium inclined-plane to PDA culture medium, back-off is cultivated；
(3) step (2) cultured mushroom mycelium is inoculated in seed culture medium and is cultivated, obtain mushroom liquor to be induced
Body strain；
(4) mushroom liquid bacterial is placed in activation induced medium and is cultivated, the activation induced medium includes following
The component of weight percent: wheat bran 0.3%, dregs of beans 0.3%, corn flour 0.2%, glucose 2%, yeast extract 0.3%, KH2PO4
0.1%, MgSO4·7H2O 0.05%, lignocellulosic 1-3%, remaining is distilled water.
2. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 1, which is characterized in that
The PDA culture medium includes the component of following weight: potato 200g, glucose 20g, agar 20g, distilled water 1000mL.
3. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 2, which is characterized in that
The condition of culture of the step (1) is to cultivate 9d at 25 DEG C, allows mycelium to cover with inclined-plane, then preservation is oblique under the conditions of 4 DEG C
Face mycelium, and periodically strain is passed on.
4. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 1, which is characterized in that
The seed culture medium includes the component of following weight: potato 200g, glucose 20g, peptone 10g, dipotassium hydrogen phosphate 5g,
Distilled water 1000mL.
5. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 4, which is characterized in that
The condition of culture of the step (3) is the static gas wave refrigerator 3d in 26 DEG C of shaking tables, then in revolving speed 150r/min and temperature is
8d is cultivated in 26 DEG C of shaking table, obtains mushroom liquid bacterial to be induced.
6. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 1, which is characterized in that
The lignocellulosic is one of Chinese scholartree lignocellulosic, pine tree lignocellulosic, poplar lignocellulosic.
7. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 6, which is characterized in that
The condition of culture of the step (4) is to weigh wheat bran, dregs of beans, corn flour, glucose, yeast extract, KH in proportion2PO4、MgSO4·
7H2O and lignocellulosic are configured to activation induced medium, the mushroom liquid bacteria that removing step (3) obtains after being mixed
Kind, 8d is cultivated in revolving speed 150r/min and in shaking table that temperature is 26 DEG C.
8. the method for addition lignocellulosic activation induction mushroom liquid bacterial according to claim 7, which is characterized in that
The mushroom liquid bacterial of inoculation accounts for the 5% of activation induced medium total amount.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110583371A (en) * | 2019-09-23 | 2019-12-20 | 贵州同辉食用菌发展有限公司 | Fermentation method of mushroom liquid strain with high activity and high stability |
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| WO2009020459A2 (en) * | 2006-08-04 | 2009-02-12 | Verenium Corporation | Glucanases, nucleic acids encoding them and methods for making and using them |
| CN106755283A (en) * | 2016-12-22 | 2017-05-31 | 菏泽学院 | The assay method of enzyme activity rule during a kind of Phellinus batch production liquid fermentation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110583371A (en) * | 2019-09-23 | 2019-12-20 | 贵州同辉食用菌发展有限公司 | Fermentation method of mushroom liquid strain with high activity and high stability |
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Application publication date: 20190517 |