CN103667169A - Method for producing Phanerochaete chrysosporium chlamydospores and preparation thereof through industrial large-scale fermentation and preparation - Google Patents
Method for producing Phanerochaete chrysosporium chlamydospores and preparation thereof through industrial large-scale fermentation and preparation Download PDFInfo
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Abstract
The invention provides a method for preparing Phanerochaete chrysosporium chlamydospores, which is characterized by comprising the following steps: (1) inoculating fermented seeds of Phanerochaete chrysosporium into a fermentation culture medium, wherein the fermented seeds are conidiophores and/or mycelia of Phanerochaete chrysosporium; and (2) performing liquid submerged fermentation on the fermentation culture medium inoculated with the fermented seeds of Phanerochaete chrysosporium, thus producing Phanerochaete chrysosporium chlamydospores. The invention also provides a method for preparing a Phanerochaete chrysosporium chlamydospore preparation and the preparation. According to the technical scheme, Phanerochaete chrysosporium chlamydospores can be simply produced in a large scale.
Description
Technical field
The present invention relates to the method for the chlamydospore preparation of a kind of chlamydospore of preparing Phanerochaete chrysosporium and preparation Phanerochaete chrysosporium, and the chlamydospore preparation that utilizes Phanerochaete chrysosporium prepared by the method.
Background technology
Phanerochaete chrysosporium (Phanerochaete chrysosporium) has the ability of the degraded xenobiontics of very strong and wide spectrum.From early 1980s < < Science > > reported first the Degradation of P.chrysosporium, just caused the extensive concern of environment circle, carry out the large quantity research to aspects such as white rot mycobiology characteristic, physicochemical property, application, become the useful tool that sewage disposal, exhaust emission processing and soil organisms are repaired.
P.chrysosporium can produce three kinds of propaguluies in its growth cycle, comprises mycelium, chlamydospore and conidium.The fermentative production of P.chrysosporium mainly adopts liquid fermenting mycelia to produce the meta-bolitess such as laccase, peroxidase at present, or adopts solid culture production conidium to prepare conidium preparation.Due to its mycelia or the difficult preservation of conidium, in storage and transport process, effective microbe mortality or activity decreased in microbial inoculum, cause the effect of product to reduce or function reduction, limited the large-scale production and application of P.chrysosporium microbial inoculum.Chlamydospore is the spore with resistivity of mycetogenetic a kind of cell walls thickening, can resist bad external environment, and as high temperature, low temperature, ultraviolet ray, pH value and other chemical substances, the shelf-lives of product is long, is easy to preserve.Therefore develop the chlamydospore preparation of P.chrysosporium, for the commercialized development of P.chrysosporium, utilize significant.
CN102140425A discloses the chlamydosporic cultural method of a kind of Phanerochaete chrysosporium, but the method adopts shake-flask culture fermentation P.chrysosporium, hypha body needs freezing rear milled processed to prepare microbial inoculum, the fermentation culture of the method is only shaking flask level, continuous amplification production process from shaking flask to large tank, also have many technical barriers, and the method operation steps is complicated, has certain distance with the technical requirements of scale operation.
Therefore, need exploitation badly a kind of simple to operate and be applicable to the chlamydosporic method of fermentor tank scale operation Phanerochaete chrysosporium.
Summary of the invention
The object of the invention is to prepare in order to overcome prior art the defect that the chlamydospore operation steps of Phanerochaete chrysosporium is complicated, be unsuitable for scale operation, the chlamydosporic method of the simple and Phanerochaete chrysosporium that can be mass-produced of a kind of working method is provided, prepare a method for the chlamydospore preparation of Phanerochaete chrysosporium, and the chlamydospore preparation that utilizes the Phanerochaete chrysosporium that the method obtains.
To achieve these goals, on the one hand, the invention provides a kind of chlamydosporic method of preparing Phanerochaete chrysosporium, wherein, the method comprises the steps:
(1) ferment-seeded of Phanerochaete chrysosporium is inoculated in fermention medium; Described ferment-seeded is conidium and/or the mycelium of Phanerochaete chrysosporium;
(2) substratum of having inoculated the ferment-seeded of described Phanerochaete chrysosporium is carried out to liquid submerged fermentation, to produce the chlamydospore of Phanerochaete chrysosporium;
Wherein, in step (1), the CaCO of the yeast powder of the carbohydrate that described fermention medium contains 0.5-3.5 % by weight, 0.1-2 % by weight, the corn steep liquor of 2-8 % by weight, 0.05-1 % by weight
3, 0.005-0.05 % by weight ZnSO
4, 0.005-0.05 % by weight MgSO
4, 0.01-0.05 % by weight MnSO
4water with surplus;
In step (2), the condition of described liquid submerged fermentation comprises: the temperature of fermentation is 25-31 ℃, and air flow is 1:0.5-1 volume: (volume minute), stirring velocity is 150-200rpm, the time of fermentation is 100-120 hour.
Second aspect, the invention provides a kind of method of preparing the chlamydospore preparation of Phanerochaete chrysosporium, and wherein, the method comprises:
(1) the chlamydosporic fermented liquid that contains Phanerochaete chrysosporium according to method preparation provided by the invention;
(2) the described chlamydosporic fermented liquid that contains Phanerochaete chrysosporium is mixed with flocculating aids, obtain mixed material;
(3) the mixed material described in step (2) is carried out to solid-liquid separation, obtain the chlamydospore preparation that contains Phanerochaete chrysosporium.
The third aspect, the invention provides the chlamydospore preparation of Phanerochaete chrysosporium prepared by method as above.
By technique scheme, realized working method simply and the chlamydosporic object of the Phanerochaete chrysosporium that can be mass-produced.
Other features and advantages of the present invention partly in detail are described the embodiment subsequently.
Embodiment
Below the specific embodiment of the present invention is elaborated.Should be understood that, embodiment described herein only, for description and interpretation the present invention, is not limited to the present invention.
First aspect, the invention provides a kind of chlamydosporic method of preparing Phanerochaete chrysosporium, and wherein, the method comprises the steps:
(1) ferment-seeded of Phanerochaete chrysosporium is inoculated in fermention medium; Described ferment-seeded is conidium and/or the mycelium of Phanerochaete chrysosporium;
(2) substratum of having inoculated the ferment-seeded of described Phanerochaete chrysosporium is carried out to liquid submerged fermentation, to produce the chlamydospore of Phanerochaete chrysosporium;
Wherein, in step (1), the CaCO of the yeast powder of the carbohydrate that described fermention medium contains 0.5-3.5 % by weight, 0.1-2 % by weight, the corn steep liquor of 2-8 % by weight, 0.05-1 % by weight
3, 0.005-0.05 % by weight ZnSO
4, 0.005-0.05 % by weight MgSO
4, 0.01-0.05 % by weight MnSO
4water with surplus;
In step (2), the condition of described liquid submerged fermentation comprises: the temperature of fermentation is 25-31 ℃, and air flow is 1:0.5-1 volume: (volume minute), stirring velocity is 150-200rpm, the time of fermentation is 100-120 hour.
Preferably, in step (1), the CaCO of the yeast powder of the carbohydrate that described fermention medium contains 1-1.5 % by weight, 0.5-1 % by weight, the corn steep liquor of 3-5 % by weight, 0.1-0.5 % by weight
3, 0.01-0.02 % by weight ZnSO
4, 0.01-0.03 % by weight MgSO
4mnSO with 0.02-0.04 % by weight
4.
In the present invention, the carbohydrate containing in fermention medium described in step (1) is this area routine variously can provide for substratum the carbohydrate of carbon source, preferably, described carbohydrate is one or more in starch, dextrose plus saccharose; The natural pH value that the pH value of described fermention medium is substratum, is preferably 6.5-6.8.
In the present invention, the source of described Phanerochaete chrysosporium has no particular limits, for example, can be in Research for Industrial Microbial Germ preservation administrative center by commercially available, for example, numbering can be CICC40299, can be also CICC40719.
According to the present invention, the inoculum size of the ferment-seeded of described Phanerochaete chrysosporium can be conventional inoculum size, and the every liter of described fermention medium of preferably take is benchmark, and the inoculum size of described ferment-seeded is 5-10 volume %.
Wherein, in step (2), the operation of described liquid submerged fermentation can be carried out according to conventional liquid submerged fermentation method, under preferable case, the operation of described liquid submerged fermentation comprises the steps: (1) inoculation step, in described inoculation step, the conidium of Phanerochaete chrysosporium as above and/or mycelium are inoculated in liquid nutrient medium; (2) fermentation step, in described fermentation step, will inoculate Phanerochaete chrysosporium conidium and/or mycelial liquid nutrient medium ferments in fermentor tank.
Wherein, the conidium of described Phanerochaete chrysosporium and/or mycelium can be used conventional slant culture to obtain, for example, the mycelium of Phanerochaete chrysosporium as above can be seeded in to PDA substratum and (contain potato 180-220g/L, glucose 18-22g/L, agar 10-25g/L, pH value 6.8-7.0) on, at 29-31 ℃, cultivate 5-7 days, can obtain being attached in a large number conidium and/or mycelium on substratum, the substratum that slant culture can be finished digs piece to obtain conidium and/or the mycelium for inoculating.Wherein, the substratum that also slant culture can be finished digs piece and is inoculated in enlarged culturing in liquid nutrient medium, obtains the mycelium of enlarged culturing, using the mycelium of enlarged culturing as the inoculum for fermentative production.
The method of described enlarged culturing is conventionally known to one of skill in the art, and for example, the substratum that slant culture can be finished digs piece and is inoculated in shake-flask culture base and (contains starch 5-15g/L, glucose 5-15g/L, yeast powder 3-7g/L, CaCO
33-5g/L, pH value 6.5-6.8) in, in temperature, be to cultivate 20-30 hour under 29-35 ℃, the rotating speed condition that is 150-250rpm, obtain a large amount of mycelium; And the described mycelium obtaining is inoculated in to seed tank culture base (contains starch 5-15g/L, glucose 5-15g/L, yeast powder 3-7g/L, CaCO
33-5g/L, pH value nature) in, in temperature, be to cultivate 20-30 hour under 29-31 ℃, the rotating speed condition that is 150-200rpm, obtain a large amount of mycelium for fermention medium inoculation.
Wherein, the condition of described fermentation can be conventional liquid submerged fermentation condition, and for example, the temperature of fermentation can be 25-31 ℃, and air flow can be 1:0.5-1 volume: (volume minute), stirring velocity can be 150-200rpm.Although under such fermentation condition, can realize the chlamydosporic object of scale operation Phanerochaete chrysosporium, but in order further to improve the chlamydosporic output of Phanerochaete chrysosporium, under preferable case, the temperature of fermenting 0-40 hour is 29-31 ℃, and the 40th hour later temperature of fermenting is 27-29 ℃; The air flow fermenting 0-10 hour is 1:0.5-0.8 volume: (volume minute), the air flow fermenting 10-40 hour is 1:0.8-1 volume: (volume minute), and the 40th hour later air flow that ferment is 1:0.5-0.8 volume: (volume minute); Stirring velocity is 175-185rpm.Fermentation period is 100-120 hour, and now, more than 80% mycelium has all formed chlamydospore.
Term " air flow " generally recently represents with ventilation, conventionally with the volume of air by unit volume nutrient solution in per minute, recently represents (V/Vmin
-1), for example ventilation ratio is 1:0.1-1, being called for short air flow is 0.1-1 volume: (volume minute).
Second aspect, the invention provides a kind of method of preparing the chlamydospore preparation of Phanerochaete chrysosporium, and wherein, the method comprises:
(1) the chlamydosporic fermented liquid that contains Phanerochaete chrysosporium according to method preparation provided by the invention;
(2) the described chlamydosporic fermented liquid that contains Phanerochaete chrysosporium is mixed with flocculating aids, obtain mixed material;
(3) the mixed material described in step (2) is carried out to solid-liquid separation, obtain the chlamydospore preparation that contains Phanerochaete chrysosporium.
Kind and the add-on of described flocculating aids have no particular limits, as long as can promote the solid-liquid separation of chlamydosporic dispersion and fermented liquid, and chlamydospore are not had to toxic action.Preferably, described flocculating aids is diatomite and/or light calcium carbonate, take every liter of fermented liquid as benchmark, and the add-on of described flocculating aids is 3-5 % by weight.Wherein, described " flocculating aids " is a kind of too closely knit for preventing filter residue accumulation, make to filter and carry out smoothly, and the different insoluble inert material of granularity using.
Wherein, the method for described solid-liquid separation is conventionally known to one of skill in the art, for example, can use filter press filtration or centrifugal method to obtain the chlamydospore preparation that contains Phanerochaete chrysosporium.
According to the present invention, preservation for the ease of the chlamydospore preparation that contains Phanerochaete chrysosporium, under preferable case, the method also comprises that solid filter cake that solid-liquid separation is obtained or centrifugal sediment are dried and pulverize and obtains the chlamydosporic dry preparation that contains Phanerochaete chrysosporium.
Wherein, described dry condition comprises: dry temperature can be 60-80 ℃; The dry time is can be 16-20 hour.
The third aspect, the invention provides the chlamydospore preparation of Phanerochaete chrysosporium prepared by method as above.
Below will describe the present invention by embodiment.
In following examples and comparative example, the method for counting by blood counting chamber is calculated sporulation quantity.
Phanerochaete chrysosporium, purchased from Research for Industrial Microbial Germ preservation administrative center, is numbered CICC40299.
Preparation example 1
Get potato 20g after peeling and boil and within 20 minutes, get juice, juice and 2g glucose, 1.5g agar and water are made into 100ml, the cooling PDA substratum that obtains after bottling sterilizing.
By 5g starch, 5g glucose, 2.5g yeast powder, 0.15g CaCO
3be mixed with 500mL with water, the cooling shake-flask culture base that obtains after bottling (every bottle of 100mL) sterilizing.
By 5kg starch, 5kg glucose, 2.5kg yeast powder, 1.5kg CaCO
3be mixed with 500L with water, pack into and in seeding tank, after sterilizing, obtain seed tank culture base.
Preparation example 2
Phanerochaete chrysosporium is inoculated on the PDA substratum in preparation example 1, cultivates 5 days at 30 ℃; By inoculate and cultivate after PDA substratum dig piece and be inoculated in the shaking flask liquid nutrient medium in preparation example 1 (the lawn 1cm of 100ml liquid nutrient medium inoculation PDA substratum growth
2), under the condition of 30 ℃ and rotating speed 200rpm, cultivate 24 hours, as seed fermenter inoculum; By in the seed tank culture base after cultured shaking flask inoculum access sterilizing (inoculum size 0.5%(v/v)), under the condition of 30 ℃ and rotating speed 180rmp, cultivate 24 hours, become ferment-seeded.
Embodiment 1
The present embodiment is for illustrating the method for the chlamydosporic method of preparation Phanerochaete chrysosporium provided by the invention and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
(1) preparation of fermention medium: the every liter of fermented liquid of take is benchmark, starch content is that 1.5 % by weight, yeast powder content are that 0.5 % by weight, corn steep liquor content are 4 % by weight, CaCO
3content is 0.3 % by weight, ZnSO
4content be 0.02 % by weight, MgSO
4content be 0.025 % by weight and MnSO
4content be that 6500 liters of substratum of 0.03 % by weight carry out sterilizing and obtain fermention medium, natural pH value.
(2) by the ferment-seeded obtaining in preparation example 2, by 5%(volume/volume) inoculum size be inoculated in the fermention medium in the present embodiment step (1).Fermentation condition in fermentor tank is: the leavening temperature fermenting 0-40 hour is 30 ℃; The 40th hour later leavening temperature that ferment is 28 ℃; The air flow fermenting 0-10 hour is 1:0.6 volume: (volume minute); The air flow fermenting 10-40 hour is 1:1 volume: (volume minute), and the 40th hour later air flow that ferment is 1:0.6 volume: (volume minute); Stirring velocity is 180rpm.
(3) fermentation is in the time of the 110th hour, and 90% above mycelia all forms chlamydospore, and fermented liquid is thinning, add 325kg diatomite, stirring and evenly mixing, passes through filter press by fermented liquid now, collect filter cake, will obtain filtration cakes torrefaction and pulverizing and obtain Phanerochaete chrysosporium chlamydospore preparation.Wherein, described dry temperature is 60 ℃, and the dry time is 20 hours.Sporulation quantity is 5.6 * 10
7individual chlamydospore/mL.
Embodiment 2
The present embodiment is for illustrating the method for the chlamydosporic method of preparation Phanerochaete chrysosporium provided by the invention and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
(1) preparation of fermention medium: the every liter of fermented liquid of take is benchmark, glucose content is that 1 % by weight, yeast powder content are that 1 % by weight, corn steep liquor content are 5 % by weight, CaCO
3content is 0.1 % by weight, ZnSO
4content be 0.01 % by weight, MgSO
4content be 0.03 % by weight and MnSO
4content be that 6500 liters of substratum of 0.04 % by weight carry out sterilizing and obtain fermention medium.
(2) by the ferment-seeded obtaining in preparation example 2, by 7%(volume/volume) inoculum size be inoculated in the fermention medium in the present embodiment step (1).Fermentation condition in fermentor tank is: the leavening temperature fermenting 0-40 hour is 31 ℃; The 40th hour later leavening temperature that ferment is 29 ℃; The air flow fermenting 0-10 hour is 1:0.8 volume: (volume minute); The air flow fermenting 10-40 hour is 1:0.9 volume: (volume minute), and the 40th hour later air flow that ferment is 1:0.5 volume: (volume minute); Stirring velocity is 185rpm.
(3) fermentation is in the time of the 110th hour, and 90% above mycelia all forms chlamydospore, and fermented liquid is thinning, add 325kg light calcium carbonate, stirring and evenly mixing, collects chlamydospore by fermented liquid now by filter press, collect filter cake, and dry and pulverize and be Phanerochaete chrysosporium chlamydospore preparation.Wherein, described dry temperature is 65 ℃, and the dry time is 19 hours.Sporulation quantity is 3.2 * 10
7individual chlamydospore/mL.
Embodiment 3
The present embodiment is for illustrating the method for the chlamydosporic method of preparation Phanerochaete chrysosporium provided by the invention and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
(1) preparation of fermention medium: the every liter of fermented liquid of take is benchmark, sucrose content is that 1.3 % by weight, yeast powder content are that 0.8 % by weight, corn steep liquor content are 3 % by weight, CaCO
3content is 0.5 % by weight, ZnSO
4content be 0.015 % by weight, MgSO
4content be 0.01 % by weight and MnSO
4content be that 6500 liters of substratum of 0.02 % by weight carry out sterilizing and obtain fermention medium.
(2) by the ferment-seeded obtaining in preparation example 2, by 10%(volume/volume) inoculum size be inoculated in the fermention medium in the present embodiment step (1).Fermentation condition in fermentor tank is: the leavening temperature fermenting 0-40 hour is 29 ℃; The 40th hour later leavening temperature that ferment is 27 ℃; The air flow fermenting 0-10 hour is 1:0.5 volume: (volume minute); The air flow fermenting 10-40 hour is 1:0.8 volume: (volume minute), and the 40th hour later air flow that ferment is 1:0.8 volume: (volume minute); Stirring velocity is 175rpm.
(3) fermentation is in the time of the 110th hour, and 90% above mycelia all forms chlamydospore, and fermented liquid is thinning, add 325kg light calcium carbonate, stirring and evenly mixing, collects chlamydospore by fermented liquid now by filter press, collect filter cake, and dry and pulverize and be Phanerochaete chrysosporium chlamydospore preparation.Wherein, described dry temperature is 80 ℃, and the dry time is 16 hours.Sporulation quantity is 2.1 * 10
7individual chlamydospore/mL.
Embodiment 4
The present embodiment is for illustrating the method for the chlamydosporic method of preparation Phanerochaete chrysosporium provided by the invention and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
According to the method for embodiment 1, carry out the preparation of the chlamydosporic preparation of Phanerochaete chrysosporium and the chlamydospore preparation of Phanerochaete chrysosporium, different is, carry out the composition of fermention medium prepared by the chlamydospore of Phanerochaete chrysosporium for take every liter of fermented liquid as benchmark, starch content is that 3.5 % by weight, yeast powder content are that 0.1 % by weight, corn steep liquor content are 8 % by weight, CaCO
3content is 0.05 % by weight, ZnSO
4content be 0.05 % by weight, MgSO
4content be 0.005 % by weight and MnSO
4content be 0.05 % by weight.Sporulation quantity is 7.1 * 10
6individual chlamydospore/mL.
Embodiment 5
The present embodiment is for illustrating the method for the chlamydosporic method of preparation Phanerochaete chrysosporium provided by the invention and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
According to the method for embodiment 1, carry out the preparation of the chlamydosporic preparation of Phanerochaete chrysosporium and the chlamydospore preparation of Phanerochaete chrysosporium, different is, the composition that carries out fermention medium prepared by the chlamydospore of Phanerochaete chrysosporium is, take every liter of fermented liquid as benchmark, and starch content is that 0.5 % by weight, yeast powder content are that 2 % by weight, corn steep liquor content are 2 % by weight, CaCO
3content is 1 % by weight, ZnSO
4content be 0.005 % by weight, MgSO
4content be 0.05 % by weight and MnSO
4content be 0.01 % by weight.Sporulation quantity is 6.6 * 10
6individual chlamydospore/mL.
Embodiment 6
The present embodiment is for illustrating the method for the chlamydosporic method of fermention medium provided by the invention, preparation Phanerochaete chrysosporium and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
According to the method for embodiment 1, carry out the preparation of the chlamydosporic preparation of Phanerochaete chrysosporium and the chlamydospore preparation of Phanerochaete chrysosporium, different, the leavening temperature fermenting 0-40 hour in the process of fermentation is 31 ℃; The 40th hour later leavening temperature that ferment is 25 ℃.Sporulation quantity is 9.2 * 10
6individual chlamydospore/mL.
Embodiment 7
The present embodiment is for illustrating the method for the chlamydosporic method of preparation Phanerochaete chrysosporium provided by the invention and the chlamydospore preparation of preparation Phanerochaete chrysosporium.
According to the method for embodiment 1, carry out the preparation of the chlamydosporic preparation of Phanerochaete chrysosporium and the chlamydospore preparation of Phanerochaete chrysosporium, different, the temperature of fermenting in the whole process of fermentation is 30 ℃.Sporulation quantity is 1.2 * 10
7individual chlamydospore/mL.
Comparative example 1
The present embodiment is for illustrating the method for the chlamydosporic method of the preparation Phanerochaete chrysosporium that prior art provides and the chlamydospore preparation of preparation Phanerochaete chrysosporium
According to the method for embodiment 7, carry out the preparation of the chlamydosporic preparation of Phanerochaete chrysosporium and the chlamydospore preparation of Phanerochaete chrysosporium, different, the temperature of fermenting in the whole process of fermentation is 33 ℃.Sporulation quantity is 2.3 * 10
6individual chlamydospore/mL.
Embodiment 1-7 by above-mentioned employing technical solution of the present invention compares with the comparative example 1 of the technical scheme that adopts prior art to provide, and adopts fermention medium of the present invention, zymotechnique, and sporulation quantity can improve greatly.Embodiment 1-3 and embodiment 4 and 5 are compared and can be found out, when in fermention medium, the content of each material is in the preferred scope of the present invention, its sporulation quantity is higher.Embodiment 1-3 and embodiment 6 and 7 are compared and can be found out, when fermentation condition is in the preferred scope of the present invention, its sporulation quantity is higher.Therefore, adopt method of the present invention can produce on a large scale the chlamydospore of the flat lead fungi of yellow archespore; And after fermentation ends, the operation scheme of chlamydospore preparation of preparing the flat lead fungi of yellow archespore is simple.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition each the concrete technical characterictic described in above-mentioned embodiment, in reconcilable situation, can combine by any suitable mode.For fear of unnecessary repetition, the present invention is to the explanation no longer separately of various possible array modes.
In addition, between various embodiment of the present invention, also can carry out arbitrary combination, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (8)
1. prepare the chlamydosporic method of Phanerochaete chrysosporium, it is characterized in that, the method comprises the steps:
(1) ferment-seeded of Phanerochaete chrysosporium is inoculated in fermention medium; Described ferment-seeded is conidium and/or the mycelium of Phanerochaete chrysosporium;
(2) fermention medium of having inoculated the ferment-seeded of described Phanerochaete chrysosporium is carried out to liquid submerged fermentation, to produce the chlamydospore of Phanerochaete chrysosporium;
Wherein, in step (1), the CaCO of the yeast powder of the carbohydrate that described fermention medium contains 0.5-3.5 % by weight, 0.1-2 % by weight, the corn steep liquor of 2-8 % by weight, 0.05-1 % by weight
3, 0.005-0.05 % by weight ZnSO
4, 0.005-0.05 % by weight MgSO
4, 0.01-0.05 % by weight MnSO
4water with surplus;
In step (2), the condition of described liquid submerged fermentation comprises: the temperature of fermentation is 25-31 ℃, and air flow is 1:0.5-1 volume: (volume minute), stirring velocity is 150-200rpm, the time of fermentation is 100-120 hour.
2. method according to claim 1, wherein, in step (1), the CaCO of the yeast powder of the carbohydrate that described fermention medium contains 1-1.5 % by weight, 0.5-1 % by weight, the corn steep liquor of 3-5 % by weight, 0.1-0.5 % by weight
3, 0.01-0.02 % by weight ZnSO
4, 0.01-0.03 % by weight MgSO
4mnSO with 0.02-0.04 % by weight
4.
3. method according to claim 1 and 2, wherein, in step (1), described carbohydrate is one or more in starch, dextrose plus saccharose.
4. method according to claim 1, wherein, in step (2), the condition of described liquid submerged fermentation comprises: the temperature of fermenting 0-40 hour is 29-31 ℃, the 40th hour later temperature of fermenting is 27-29 ℃; The air flow fermenting 0-10 hour is 1:0.5-0.8 volume: (volume minute), the air flow fermenting 10-40 hour is 1:0.8-1 volume: (volume minute), and the 40th hour later air flow that ferment is 1:0.5-0.8 volume: (volume minute); Stirring velocity is 175-185rpm.
5. a method of preparing the chlamydospore preparation of Phanerochaete chrysosporium, is characterized in that, the method comprises:
(1) the chlamydosporic fermented liquid that contains Phanerochaete chrysosporium according to the method preparation described in any one in claim 1-4;
(2) the described chlamydosporic fermented liquid that contains Phanerochaete chrysosporium is mixed with flocculating aids, obtain mixed material;
(3) the mixed material described in step (2) is carried out to solid-liquid separation, obtain the chlamydospore preparation that contains Phanerochaete chrysosporium.
6. method according to claim 5, wherein, with respect to the chlamydosporic fermented liquid that contains Phanerochaete chrysosporium described in every weight part, the add-on of flocculating aids is 3-5 % by weight.
7. according to the method described in claim 5 or 6, wherein, described flocculating aids is diatomite and/or light calcium carbonate.
8. the chlamydospore preparation of the Phanerochaete chrysosporium that in claim 5-7 prepared by the method described in any one.
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|---|---|---|---|
| CN201310228809.5A CN103667169B (en) | 2013-06-08 | 2013-06-08 | Method and the preparation thereof of Phanerochaete chrysosporium chlamydospore and preparation thereof are produced in industrial scale fermentation |
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| CN105543159A (en) * | 2016-03-02 | 2016-05-04 | 河南师范大学 | Method for producing CLCs (chlamydospore-like cells) through solid fermentation of corn straw by using Phanerochaete chrysosporium |
| CN105884039A (en) * | 2016-05-11 | 2016-08-24 | 河南师范大学 | Microbial curing bed reactor for treating highly concentrated phenol-containing wastewater |
| CN106591276A (en) * | 2016-12-23 | 2017-04-26 | 南京理工大学 | Method for degrading malachite green by using bacterial cellulose membrane-immobilized phanerochaete chrysosporium |
| CN111925948A (en) * | 2020-08-14 | 2020-11-13 | 四川金珠生态农业科技有限公司 | Preparation method and application of high-concentration spore liquid of Ceriporia lacerata |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543159A (en) * | 2016-03-02 | 2016-05-04 | 河南师范大学 | Method for producing CLCs (chlamydospore-like cells) through solid fermentation of corn straw by using Phanerochaete chrysosporium |
| CN105884039A (en) * | 2016-05-11 | 2016-08-24 | 河南师范大学 | Microbial curing bed reactor for treating highly concentrated phenol-containing wastewater |
| CN105884039B (en) * | 2016-05-11 | 2019-10-15 | 河南师范大学 | It is a kind of for handling the microorganism fixed bedreactor of high concentration phenol waste water |
| CN106591276A (en) * | 2016-12-23 | 2017-04-26 | 南京理工大学 | Method for degrading malachite green by using bacterial cellulose membrane-immobilized phanerochaete chrysosporium |
| CN111925948A (en) * | 2020-08-14 | 2020-11-13 | 四川金珠生态农业科技有限公司 | Preparation method and application of high-concentration spore liquid of Ceriporia lacerata |
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| CN103667169B (en) | 2015-08-05 |
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