CN106635823A - Method for controlling growth morphology of mycelia - Google Patents
Method for controlling growth morphology of mycelia Download PDFInfo
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Abstract
The invention relates to the technical field of bioengineering and particularly relates to a method for controlling the growth morphology of mycelia. The method comprises the following steps: at a mycelium growth phase of a filamentous fungus fermentation process, adding farnesol into a fermented solution; and continually fermenting to obtain spherical filamentous fungus mycelia, wherein the final concentration of added farnesol is 150-1000 microns. According to the method provided by the invention, the growth morphology in the filamentous fungus fermentation process can be efficiently and conveniently controlled and the fermentation yield of filamentous fungi is improved; and the method is simple to operate and high in universality and is suitable for being amplified in industrial production.
Description
Technical field
The present invention relates to technical field of bioengineering, more particularly to a kind of method of control mycelial growth form.
Background technology
Filamentous fungis have a wide range of applications in industrialized production, and product covers industrial enzyme preparation (such as amylase, fibre
The plain enzyme of dimension, protease, lipase etc.), organic acid, antibiotic etc..In industrialization production, the filamentous fungis of submerged fermentation are logical
There are 3 kinds of forms in the case of often：It is lumps, cotton-shaped and spherical.The filamentous fungis of different shape can be directly affected in fermentation system
Physical parameter, substrate transmission etc., so generation of the growthform to fermented product, the control of sweat is most important.It is thread
The growthform of funguses has a significant impact to cellular metabolic pathways, product accumulation and separation and Extraction, therefore how to control thread
The growthform of funguses, maximizes the emphasis for improving target product yield into commercial production research.
During the fermentation inoculum concentration is difficult to control crumby thalli morphology, and significantly limit the biography inside thalline
Oxygen, mass transfer, Product yields are substantially low, and in stirred-tank fermenter sweat, can be wrapped on agitator or attached
Wall, ultimately resulting in can not obtain target product, fermentation failure；Cotton-shaped thalline nutrient substance and oxygen are more suitable in its internal transmission
Profit, but it can increase fermentation liquid viscosity, and rheological characteristic is relatively poor, reduces the transmission efficiency of material, oxygen in fermentation liquid, while
Stirring paddle is also easily wound, fermentation product production, fermentation tank performance is reduced；Spherical-like morphology can overcome the above not enough, improve
The rheological behavior of fermentation liquid, effectively improves mass transfer, passes oxygen performance, reducing energy consumption, if but fungus ball is excessive, fungus ball internal mass transfer, biography
Oxygen will occur difficult, impact product accumulation, therefore rule, suitable size spherical funguses are then conducive to enzyme preparation, organic acid
Deng the production of product.
However, growthform develops a kind of instinct form of expression as filamentous fungis life entity, the factor that is affected is more
And process is complicated, such as the impact of strain itself, the impact of growing nutrient environment, the impact of fermentation operation condition etc., most
The form for causing filamentous fungis eventually not can be controlled to preferable form, or the repeatability and collimation of fungus ball control are bad.
This Morphology Control Technology for allowing for high-efficient simple seems and is even more important.In filamentous fungis sweat, different metabolic product
Suitable form it is different, under different shape, the Accumulation discrepancy of metabolite substantially, the spherical head for becoming many productions
Form slection state, in today that immobilization technology is developed rapidly, control thalli growth becomes the self-curing technology of fungus ball and has obtained huge
Development.
Environment is the key factor for affecting fungus ball to be formed, the fungus ball structure and different sizes of acquisition under the conditions of varying environment.
Affect that fungus ball is formed and the factor of characteristic typically has pH, inoculating spores, medium component, speed of agitator etc., by these because
The regulation and control of element can obtain the optimal morphology of product accumulation.
Liu Y,Liao W,Chen SL.“Study of pellet formation of filamentous fungi
Rhizopus oryzae using a multiple logistic regression model.”Biotechnol
Bioeng,2008,99(1):Report in 117-128.：Integrated survey spore concentration, glucose, carbamide, phosphate, metal
Impact of the concentration, fermentation system pH, ventilation stirring of ion to filamentous fungis growthform, pH and inoculum concentration are to affect thread true
The main cause of bacterium form, by reducing embracing sub- concentration, strict modification scope is effectively controlled to filamentous fungis spherical.The skill
The strategy of art scheme is：By changing growth of microorganism environment, suppress the growth of microorganism, be finally reached control pellet form
Purpose.It is often required that and for acidity adjustment to be accurate to 2 significant digits the disadvantage is that, require the regulation of pH very high, and this makes
Must be difficult to amplify in the industrial production.In addition, the certain Acidity Range of the characteristic requirements of some products, also makes the technical scheme
Using being restricted.
Driouch H,Sommer B,Wittmann C.“Morphology engineering of Aspergillus
niger for improved enzyme production.”Biotechnol Bioeng,2010,105(6):1058-
1068. in report：Can be used as the available strategy of morphology Control with the effect of production bacterial strain in flow field by the use of micropartical.
When using aspergillus niger producing enzyme, the micropartical such as silicon dioxide are added in fermentation system, effectively by the form of microorganism system
It is controlled to thread, has reached the purpose of high yield.The premise of its technical scheme is that the strain of the type is due to the growth spy of itself
Property, can not well form fungus ball because of the change of environment.In different fermentation systems, organic acid and some enzyme systems are carried out
During the fermenting and producing of the products such as agent, spherical is optimal form of production, add atomic technology path can not effectively by
Filamentous fungis are controlled to pellet form.
In sum, prior art processes condition is harsh, and operational approach is complicated, it is impossible to solves effective control filamentous fungis and sends out
The problem of ferment form.Therefore, to control filamentous fungis Fermentative growth form, becoming necessarily needs to design efficiently easily technical scheme
Ask.
The content of the invention
It is an object of the invention to provide a kind of method of control mycelial growth state, methods described can be efficiently convenient
Growthform in ground control filamentous fungis sweat, and the method is simple to operate, universality is strong, suitably in the industrial production
Amplify.
To reach this goal of the invention, the present invention is employed the following technical solutions：
In a first aspect, the invention provides a kind of method of control mycelial growth form, methods described includes：Thread
In the mycelial growth stage of fungi fermentation process, add farnesol in fermentation liquid, and it is spherical thread to obtain to continue to ferment
Fungal mycelium；
Wherein, final concentration of 150-1000 μM of the farnesol addition.
Final concentration of 150 μM of farnesol addition, 160 μM, 170 μM, 180 μM, 190 μM, 200 μM, 210 μM, 220
μM、230μM、240μM、250μM、260μM、270μM、280μM、290μM、300μM、310μM、320μM、330μM、340μM、
350μM、360μM、370μM、380μM、390μM、400μM、430μM、450μM、480μM、500μM、530μM、550μM、580μ
M、600μM、630μM、650μM、680μM、700μM、730μM、750μM、780μM、800μM、830μM、850μM、880μM、900
μM, 930 μM, 950 μM, 980 μM or 1000 μM,.
In the present invention, used as widest quorum sensing molecule is studied in Fungal quorum-sensing system, it can have farnesol
Effect changes the thalli morphology of Candida albicans, suppresses it from Yeast protoplase to the transformation of hypha form, therefore, by debita spissitudo
Farnesol is added to as additives can be with effective control filamentous fungis thalli morphology in fermentation medium.
In the present invention, the farnesol solution of certain concentration is added in filamentous fungis fermentation medium, by filamentous fungis
Mycelium morphology is controlled to spherical.
Preferably, 180-800 μM of the final concentration of the farnesol addition, preferably 200-600 μM.
Preferably, farnesol is added in fermentation liquid in the form of farnesol solution, the preparation of the farnesol solution is molten
Agent is the one kind in ethanol, normal propyl alcohol, isopropanol and n-butyl alcohol or at least two mixing, preferably ethanol or isopropanol.
Preferably, the filamentous fungis are any one in aspergillus, Rhizopus, trichoderma or Trametes or at least two
The combination planted, preferably any one in aspergillus, Rhizopus or trichoderma or at least two combination.
Preferably, the method for the control mycelial growth form, specifically includes following steps：
(1) pre-incubated seed culture fluid is inoculated in fermentation medium, carries out fermentation culture；
(2) in the mycelial growth stage of the fermentation culture process, the additive process Buddhist nun alcoholic solution in fermentation liquid；
(3) collects thalline after mycelial growth terminates, observes mycelial growth form.
Preferably, fermentation medium includes following component in the step (1)：Carbon source 1-50g/L, nitrogen source 1-20g/L,
KH2PO40.5-5g/L, MgSO4·7H2O 0.1-2g/L, FeSO4·7H2O 0.01-0.1g/L, CaCl2·2H2O 0.01-
0.1g/L。
Preferably, the carbon source is any one in glycerol, glucose or sucrose or at least two mixing.
Preferably, the nitrogen source is soy peptone, carbamide or (NH4)2SO4In any one or at least two it is mixed
Close.
Preferably, the seed culture fluid is inoculated into fermentation medium, seed culture fluid inoculum concentration by volume
For 3-15%, for example can be 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or
15%, preferably 4-12%, more preferably 6-10%.
Preferably, the pH of the fermentation culture is 4.0-6.0, for example can be 4.0,4.1,4.2,4.3,4.4,4.5,
4.6th, 4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4,5.5,5.6,5.7,5.8,5.9 or 6.0, preferably 4.5-5.5.
Preferably, the temperature of the fermentation culture is 20-40 DEG C, for example can be 20 DEG C, 21 DEG C, 22 DEG C, 23 DEG C, 24
℃、25℃、26℃、27℃、28℃、29℃、30℃、31℃、32℃、33℃、34℃、35℃、36℃、37℃、38℃、39℃
Or 40 DEG C, preferably 23-36 DEG C, more preferably 26-32 DEG C.
Preferably, the stir speed (S.S.) of the fermentation culture be 50-200rpm, for example can be 50rpm, 60rpm, 70rpm,
80rpm、90rpm、100rpm、110rpm、120rpm、130rpm、140rpm、150rpm、160rpm、170rpm、180rpm、
190rpm or 200rpm, preferably 100-180rpm.
Preferably, the time of the fermentation culture be 48-168h, for example can be 48h, 50h, 52h, 54h, 55h, 58h,
60h、62h、65h、68h、70h、72h、75h、78h、80h、82h、85h、88h、90h、92h、95h、98h、100h、102h、
105h、108h、110h、112h、115h、118h、120h、122h、125h、128h、130h、135h、138h、140h、142h、
145h, 148h, 150h, 155h, 158h, 160h, 162h, 165h or 168h.
Preferably, the step (2) to additive process Buddhist nun alcoholic solution in fermentation liquid, the farnesol is for disposably or several times
Addition, preferably disposable addition.
The farnesol solution is degerming using 0.22 μm of membrane filtration.
Preferably, the concentration of the farnesol solution is 200-2000 times of the final concentration of the addition, for example, can be
Such as 200 times, 400 times, 600 times, 800 times, 1000 times, 1200 times, 1400 times, 1600 times, 1800 times, 2000 times, preferably
500-1500 times, more preferably 800-1200 times.
Preferably, the addition time of the farnesol solution is the 0-48h of fermentation culture process in the step (1),
For example can be 0h, 2h, 4h, 6h, 8h, 10h, 12h, 14h, 16h, 18h, 20h, 22h, 24h, 26h, 28h, 30h, 32h, 34h,
36h, 38h, 40h, 42h, 44h, 46h, 48h, preferably 4-36h, more preferably 12-24h.
Preferably, during the fermentation culture mycelial growth time be 0-48h, for example can be 0h, 2h, 4h,
6h、8h、10h、12h、14h、16h、18h、20h、22h、24h、26h、28h、30h、32h、34h、36h、38h、40h、42h、
44h, 46h, 48h, preferably 12-40h, more preferably 20-36h.
As optimal technical scheme, the method for the control mycelial growth form, following steps are specifically included：
(1 '), will be pre-incubated thread under conditions of temperature 20-40 DEG C, pH4.0-6.0, stir speed (S.S.) 50-200rpm
Funguses seed liquor is inoculated in fermentation medium with volume ratio 3-15%, carries out fermentation culture；
(2 ') the fermentation culture process 0-48h, disposably to additive process Buddhist nun alcoholic solution in fermentation liquid to method Buddhist nun
Final concentration of 110-400 μM of alcohol；
(3 ') collects thalline after mycelial growth terminates, observes mycelial growth form.
Compared with prior art, the invention has the advantages that：
(1) method of the control mycelial growth form that the present invention is provided, efficiently can easily control filamentous fungis and send out
Growthform during ferment, improves the fermentation production rate of filamentous fungis, and lovastatin content improves 1.6 times, and extracellular polysaccharide contains
Amount improves 2.1 times；
(2) method of the present invention is simple to operate, it is easy to amplifies production, large scale fermentation is suitable to, so as to effective control is thread
The form of mycothallus, meets the requirement of scale application.
Description of the drawings
Fig. 1 is the aspect graph of the bread mold of the culture of the embodiment of the present invention 1；
Fig. 2 is the aspect graph of the aspergillus niger of the culture of the embodiment of the present invention 2.
Specific embodiment
Technical scheme is further illustrated below by specific embodiment.Those skilled in the art should be bright
, the embodiment be only to aid in understand the present invention, be not construed as to the present invention concrete restriction.
Experimental technique used in following embodiments if no special instructions, is conventional method.
Material used, reagent etc. in following embodiments, if no special instructions, commercially obtain.
Strain：The strain of the present embodiment is the filamentous fungis with Typical Representative commonly used in commercial production：
Bread mold (Rhizopus nigricans 41346) spore suspension；
Aspergillus niger (Aspergillus niger 2106) spore suspension；
Monascuses (Monascus purpureus 5013) spore suspension；
Trametes versicolor (Trametes versicolor 14001).
Embodiment 1
This example demonstrates that the method for controlling mycelium morphology using farnesol：
(1) preparation of culture medium：Basal medium includes, glucose 20g/L, (NH4)2SO45g/L, KH2PO44g/L,
MgSO4·7H2O 2g/L,FeSO4·7H2O 0.1g/L,CaCl2·2H2O 0.5g/L, glucose separately sterilizes, and 115 DEG C go out
Bacterium 30min；
(2) seed culture：Bread mold spore suspension 1mL, 28 DEG C, 160rpm are added toward step (1) culture medium
Culture 20 hours, will be seed liquor after culture homogenizing；
(3) farnesol is dissolved in ethanol by the concentration of 200mM, 0.22 μm of membrane filtration is degerming, farnesol is obtained molten
Liquid；
(4) concentration in the 0h of whole incubation, addition method Buddhist nun alcoholic solution to fermentation liquid reaches 200 μM, is not added with
Enter the culture medium of farnesol as control；
(5) fermentation culture：Seed liquor is accessed into step (1) fresh culture by 6% (v/v) inoculum concentration, 28 DEG C,
160rpm is cultivated 48 hours, collects thalline.
As a result as shown in figure 1, showing that mycelium is smooth spherical in surface in the culture medium for adding farnesol.
Embodiment 2
This example demonstrates that a kind of method that utilization farnesol controls mycelium morphology：
(1) preparation of culture medium：Basal medium includes, sucrose 50g/L, carbamide 10g/L, KH2PO41g/L, MgSO4·
7H2O 1g/L, CaCl2·2H2O 0.05g/L, glucose separately sterilizes, 115 DEG C of sterilizing 30min；
(2) seed culture：Aspergillus niger spores suspension 1mL, 30 DEG C, 150rpm are added toward step (1) culture medium
Culture 36 hours, will be seed liquor after culture homogenizing；
(3) farnesol is dissolved in ethanol by the concentration of 200mM, 0.22 μm of membrane filtration is degerming, farnesol is obtained molten
Liquid；
(4) concentration in the 0h of whole incubation, addition method Buddhist nun alcoholic solution to fermentation liquid reaches 150 μM, is not added with
Enter the culture medium of farnesol as control, be 6.0 with HCl regulation and control pH；
(5) fermentation culture：Seed liquor is accessed into step (1) fresh culture by 6% (v/v) inoculum concentration, 30 DEG C,
200rpm is cultivated 72 hours, collects thalline.
As a result as shown in Fig. 2 showing that mycelium is the spherical of surface hirsutism in the culture medium for adding farnesol.
Embodiment 3
This example demonstrates that a kind of carry out lovastatin fermenting and producing using farnesol control mycelial growth morphological method：
(1) preparation of culture medium：Basal medium includes, glycerol 5%, glucose 10g/L, soy peptone 2g/L,
(NH4)2SO42g/L, KH2PO41.5g/L, MgSO4·7H2O 1g/L,FeSO4·7H2O0.05g/L, CaCl2·2H2O
0.05g/L, glucose separately sterilizes, 115 DEG C of sterilizing 30min；
(2) seed culture：Monascus spore suspension 1mL, 30 DEG C, 200rpm are added toward step (1) culture medium
Culture 48 hours, will be seed liquor after culture homogenizing；
(3) farnesol is dissolved in ethanol by the concentration of 200mM, 0.22 μm of membrane filtration is degerming, farnesol is obtained molten
Liquid；
(4) in the 48h of whole incubation, the concentration in the farnesol solution to fermentation liquid in step (3) is added to reach
To 500 μM, the culture medium of farnesol is added without as control；
(4) fermentation culture：Seed liquor is accessed into step (1) fresh culture by 10% (v/v) inoculum concentration, 30 DEG C,
200rpm is cultivated 21 days, obtains the smooth spherical thallus in surface, and the cell thalline can serve as fermenting and producing lovastatin；
(5) extraction and detection of lovastatin：Take 1 volume homogenate fermentation liquid and 4 volumes absolute methanol, 30 DEG C,
200rpm shaking table mechanical shaking extraction 3h, 12000rpm are centrifuged 10min, supernatant Jing 0.22um membrane filtrations, HPLC sample detections.
HPLC testing conditions：Chromatographic column 250mm × 4.6mm C18 posts；UV-detector：Detection wavelength 237nm；Mobile phase
For 70% acetonitrile (chromatograph pure phosphoric acid adjusts pH to 2.6-2.8)；Flow velocity 0.6ml/min；Sample size 10uL；
As a result show：Lovastatin content in fermentation liquid is 0.64g/L, and lovastatin content is 0.4g/ in matched group
L。
Embodiment 4
This example demonstrates that a kind of carry out Exopolysaccharide Production From The Fermentation production using farnesol control mycelial growth morphological method：
(1) preculture：The culture medium of preculture, wherein Rhizoma Solani tuber osi are prepared using conventional Rhizoma Solani tuber osi culture medium compound method
200g/L, glucose 20g/L；Trametes versicolor mycelium is inoculated in the culture medium, is cultivated under the conditions of 26 DEG C, 150r/min
7 days；Fresh culture will be accessed after culture homogenizing by 10% (v/v) inoculum concentration, 26 DEG C, cultivate 5 days under the conditions of 150r/min,
Obtain final product pre-incubated seed liquor.
(2) preparation of fermentation medium：Glucose 40g/L, soy peptone 10g/L, KH2PO42g/L, MgSO4·
7H2O 2g/L, glucose separately sterilizes, 115 DEG C of sterilizing 30min；
(3) farnesol is dissolved in ethanol by the concentration of 200mM, 0.22 μm of membrane filtration is degerming, farnesol is obtained molten
Liquid；
(4) concentration in the 0h of whole incubation, addition method Buddhist nun alcoholic solution to fermentation liquid reaches 1mM, is added without
The culture medium of farnesol is used as control；
(5) fermentation culture：Seed liquor is accessed into step (2) fresh culture by 10% (v/v) inoculum concentration, 26 DEG C,
50rpm is cultivated 168 hours, obtains the brushy spherical thallus for expanding in surface, and the cell thalline can serve as fermenting and producing born of the same parents
Exo polysaccharides；
(6) extraction and detection of extracellular polysaccharide：The fermentation liquid of 1 volume and 95% ethanol of 4 volumes are taken, vibration shakes up, 4 DEG C
Left undisturbed overnight, 12000rpm centrifugation 10min, the ethanol of precipitate Jing 95%, 70% ethanol are respectively washed twice, and are obtained final product thick extracellular many
Sugar.
Detection：Phend-sulphuric acid detection total sugar content, the content of reducing sugar of DNS methods detection doping, extracellular polysaccharide content=
Total sugar content-content of reducing sugar；
As a result show：Extracellular polysaccharide content in fermentation liquid is 2.5g/L, and extracellular polysaccharide content is 0.81g/ in matched group
L, compares and improves 2.1 times.
Comparative example 1
The comparative example the difference is that only that the addition method Buddhist nun alcoholic solution is to the concentration in fermentation liquid in embodiment 1
120 μM are reached, in addition, remaining reagent and reagent dosage and cultural method are same as Example 1.
As a result show that mycelium is cotton-shaped in culture medium.
Comparative example 2
The comparative example the difference is that only that the addition method Buddhist nun alcoholic solution is to the concentration in fermentation liquid in embodiment 2
100 μM are reached, in addition, remaining reagent and reagent dosage and cultural method are same as Example 2.
As a result show that mycelium is cluster-shaped in culture medium.
Comparative example 3
The comparative example the difference is that only that the addition method Buddhist nun alcoholic solution is to the concentration in fermentation liquid in embodiment 3
60 μM are reached, in addition, remaining reagent and reagent dosage and cultural method are same as Example 3.
As a result show：Lovastatin content in fermentation liquid is 0.5g/L.
Comparative example 4
The comparative example the difference is that only that the addition method Buddhist nun alcoholic solution is to the concentration in fermentation liquid in embodiment 4
60 μM are reached, in addition, remaining reagent and reagent dosage and cultural method are same as Example 4.
As a result show：The content of the extracellular polysaccharide in fermentation liquid is 1.2g/L.
Applicant states that the present invention illustrates the process of the present invention, but the present invention not office by above-described embodiment
It is limited to above-mentioned processing step, that is, does not mean that the present invention has to rely on above-mentioned processing step and could implement.Art
Technical staff it will be clearly understood that any improvement in the present invention, the equivalence replacement and auxiliary element to raw material selected by the present invention
Addition, selection of concrete mode etc., within the scope of all falling within protection scope of the present invention and disclosure.
Claims (10)
1. it is a kind of control mycelial growth form method, it is characterised in that methods described includes：In filamentous fungis sweat
The mycelial growth stage, add farnesol in fermentation liquid, and continue fermentation to obtain spherical filamentous fungis mycelium；
Wherein, final concentration of 150-1000 μM of the farnesol addition.
2. method according to claim 1, it is characterised in that final concentration of 180-800 μM of the farnesol addition, it is excellent
Elect 200-600 μM as.
3. method according to claim 1 and 2, it is characterised in that added in fermentation liquid in the form of farnesol solution
Farnesol, the solvent of preparing of the farnesol solution is the one kind or at least two in ethanol, normal propyl alcohol, isopropanol and n-butyl alcohol
Mixing, preferably ethanol or isopropanol.
4. the method according to any one of claim 1-3, it is characterised in that the filamentous fungis are aspergillus, rhizopus
In category, trichoderma or Trametes any one or at least two combination, preferably in aspergillus, Rhizopus or trichoderma
Any one or at least two combination.
5. the method according to any one of claim 1-3, it is characterised in that methods described specifically includes following steps：
(1) pre-incubated seed culture fluid is inoculated in fermentation medium, carries out fermentation culture；
(2) in the mycelial growth stage of the fermentation culture process, the additive process Buddhist nun alcoholic solution in fermentation liquid；
(3) collects thalline after mycelial growth terminates, observes mycelial growth form.
6. method according to claim 5, it is characterised in that fermentation medium includes following component in the step (1)：
Carbon source 1-50g/L, nitrogen source 1-20g/L, KH2PO40.5-5g/L, MgSO4·7H2O 0.1-2g/L, FeSO4·7H2O 0.01-
0.1g/L, CaCl2·2H2O 0.01-0.1g/L；
Preferably, the carbon source is any one in glycerol, glucose or sucrose or at least two mixing；
Preferably, the nitrogen source is soy peptone, carbamide or (NH4)2SO4In any one or at least two mixing；
Preferably, the seed culture fluid is inoculated into fermentation medium, and seed culture fluid inoculum concentration by volume is 3-
15%, preferably 4-12%, more preferably 6-10%.
7. the method according to claim 5 or 6, it is characterised in that the pH of the fermentation culture is 4.0-6.0, preferably
4.5-5.5；
Preferably, the temperature of the fermentation culture is 20-40 DEG C, preferably 23-36 DEG C, more preferably 26-32 DEG C；
Preferably, the stir speed (S.S.) of the fermentation culture is 50-200rpm, preferably 100-180rpm；
Preferably, the time of the fermentation culture is 48-168h.
8. the method according to any one of claim 5-7, it is characterised in that the step (2) is added in fermentation liquid
Farnesol solution, the farnesol is preferably disposable to add disposably or several times to add；
Preferably, the concentration of the farnesol solution is 200-2000 times of the final concentration of the addition, preferably 500-1500
Times, more preferably 800-1200 times.
9. the method according to any one of claim 5-8, it is characterised in that the addition time of the farnesol solution is
The 0-48h, more preferably preferably 4-36h, 12-24h of fermentation culture process in the step (1)；
Preferably, during the fermentation culture mycelial growth time be 0-48h, preferably 12-40h, further preferably
For 20-36h.
10. the method according to any one of claim 5-9, it is characterised in that methods described specifically includes following steps：
(1 ') under conditions of temperature 20-40 DEG C, pH4.0-6.0, stir speed (S.S.) 50-200rpm, by pre-incubated filamentous fungis
Seed liquor is inoculated in fermentation medium with volume ratio 3-15%, carries out fermentation culture；
(2 ') the fermentation culture process 0-48h, disposably to additive process Buddhist nun alcoholic solution in fermentation liquid to farnesol
Final concentration of 110-400 μM；
(3 ') collects thalline after mycelial growth terminates, observes mycelial growth form.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610911793.1A CN106635823B (en) | 2016-10-19 | 2016-10-19 | Method for controlling growth form of mycelium |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113088456A (en) * | 2021-03-31 | 2021-07-09 | 南京师范大学 | Method for controlling growth form of filamentous fungi |
| CN113215206A (en) * | 2021-04-26 | 2021-08-06 | 天津科技大学 | Preparation and purification method of grifola frondosa polysaccharide with high antioxidant activity |
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| WO2006015726A1 (en) * | 2004-08-03 | 2006-02-16 | Henkel Kommanditgesellschaft Auf Aktien | Prebiotic intimate personal hygiene product |
| CN102212484A (en) * | 2011-05-05 | 2011-10-12 | 南京工业大学 | Method for controlling growthform of filamentous fungi during fermentation process |
| CN103409381A (en) * | 2013-07-19 | 2013-11-27 | 中国科学院过程工程研究所 | Method for improving Trametes versicolor laccase output |
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| WO2006015726A1 (en) * | 2004-08-03 | 2006-02-16 | Henkel Kommanditgesellschaft Auf Aktien | Prebiotic intimate personal hygiene product |
| CN102212484A (en) * | 2011-05-05 | 2011-10-12 | 南京工业大学 | Method for controlling growthform of filamentous fungi during fermentation process |
| CN103409381A (en) * | 2013-07-19 | 2013-11-27 | 中国科学院过程工程研究所 | Method for improving Trametes versicolor laccase output |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113088456A (en) * | 2021-03-31 | 2021-07-09 | 南京师范大学 | Method for controlling growth form of filamentous fungi |
| CN113215206A (en) * | 2021-04-26 | 2021-08-06 | 天津科技大学 | Preparation and purification method of grifola frondosa polysaccharide with high antioxidant activity |
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