CN101538604A - Online production method of rhamnolipid biosurfactant in cellulose hydrolyzation - Google Patents
Online production method of rhamnolipid biosurfactant in cellulose hydrolyzation Download PDFInfo
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- CN101538604A CN101538604A CN200910049013A CN200910049013A CN101538604A CN 101538604 A CN101538604 A CN 101538604A CN 200910049013 A CN200910049013 A CN 200910049013A CN 200910049013 A CN200910049013 A CN 200910049013A CN 101538604 A CN101538604 A CN 101538604A
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Abstract
The invention relates to an online production method of rhamnolipid biosurfactant in cellulose hydrolyzation, belonging to the field of biotechnology. Firstly, reaped straws are cleaned, cut and dried to constant weight, and then the cut straws are treated by alkali before cellulose hydrolyzation; secondly, the pretreated straws are cultured with Trichoderma reesei ZM4-F3 in a shaking table for hydrolyzation and sugar production for producing reducing sugar; thirdly, pseudomonas aeruginosa BSZ-07 during logarithmic phase obtained by culture is inoculated into a hydrolyzation system, thus realizing the online production of rhamnolipid biosurfactant and accelerating the hydrolyzation of cellulose. Owing to the addition method of the online production, the complex process of rhamnolipid purification can be saved, thus effectively reducing the production cost of rhamnolipid biosurfactant and being applicable for industrial production. Besides, the addition of rhamnolipid biosurfactant can improve the hydrolyzation efficiency of cellulose effectively.
Description
Technical field
The present invention relates to a kind of production method of tensio-active agent of biological technical field, specifically is the online production method of rhamnolipid biological surface activator in a kind of cellulase hydrolysis.
Background technology
Microorganism is oozy in metabolic process to have surfactivity and interfacial activity, collect hydrophilic group and hydrophobic group structure in same intramolecular amphiphilic cpds, is called bio-surfactant.Bio-surfactant comprises many different kinds, as glycolipid, lipopeptid, polysaccharide-composite of lipid, phosphatide, lipid acid and neutral fat etc.Bio-surfactant can promote the dispersion and the absorption of insoluble organic in the soil, can increase the contact surface between hydrophobic nature organic compound and the liquid phase, thereby strengthens microorganism active, improves cellulose degradation rate and composting efficiency.In addition, bio-surfactant also can be brought into play great effect in petroleum industry (as the biological viscosity reduction of oil and improve oil recovery factor etc.).And, compare with the tensio-active agent of chemosynthesis, bio-surfactant is except that having identical characteristics such as reduction surface tension, stable emulsion and foaming, also have synthetic surfactant not available nontoxic, can biological degradation etc. characteristic and bio-compatibility, therefore, people are increasing to the research of bio-surfactant in recent years.But the major obstacle that influences the bio-surfactant industrial application is its low yield and expensive.
Find through retrieval prior art, the Chinese patent publication number is CN1657613, patent name is " utilizing bio-surfactant to improve the method for viride cellulase activity ", this patent is earlier with the separation and purification of rhamnolipid type biological surfactant, be added into then in the solid-state of viride or the liquid state fermentation substratum, can improve cellulase activity.But the method that this first separation and purification is added again can increase the production cost of rhamnolipid greatly.
In the cellulase hydrolysis system, the correlation technique report of the online production addition manner that adopts bio-surfactant is not arranged so far.Because the online production of bio-surfactant can be saved the separation purifying technique of bio-surfactant, both can effectively reduce its production cost, also can effectively improve cellulosic enzymic hydrolysis efficient.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, the online production method of rhamnolipid biological surface activator in a kind of cellulase hydrolysis is provided, Trichodermareesei ZM4-F3 and Pseudomonas aeruginosa BSZ-07 are respectively applied for the production of cellulase and the production of rhamnolipid type biological surfactant.Trichodermareesei ZM4-F3 and Pseudomonas aeruginosa BSZ-07 can be bought by Chinese industrial microbial strains preservation center (CICC) and Chinese typical culture collection center (CCTCC) respectively.Pseudomonas aeruginosa BSZ-07 is added directly to in the cellulolytic system of Trichodermareesei ZM4-F3, thus the online production mode of realization rhamnolipid type biological surfactant.The online production adding technique of this kind rhamnolipid can reach and reduce production costs, and simplifies the purpose of technical process.
The present invention is achieved by the following technical solutions, comprises the steps:
The first step is cleaned harvested straw, cut off, and is dried to constant weight, before cellulase hydrolysis, the straw that cuts is handled with alkali;
Described harvested straw is clean, cut-out, be dried to constant weight, be meant: harvested straw is washed 4-5 time with removal impurity and stem knot with tap water, and cut segment to 1-2cm, the straw that cuts places 70 ℃ of baking ovens to be dried to constant weight, at room temperature stores.
Describedly will shear good straw and handle with alkali, and be meant: the straw that 10g is sheared places the sodium hydroxide of 40ml 2% (volumetric concentration v/v), and keeping solid-liquid body mass ratio is 1: 4,85 ℃ of water-bath 1h.Be washed till neutrality with distilled water, place 70 ℃ of baking ovens to be dried to constant weight.
Second goes on foot, and the straw after handling is hydrolyzed with Trichodermareesei ZM4-F3 produces the sugar cultivation, in order to the generation reducing sugar.
Described being hydrolyzed with Trichodermareesei ZM4-F3 produced sugar and cultivated, and is to carry out in 30 ℃, 200rpm shaking table.
Described Trichodermareesei ZM4-F3 hydrolysis is produced sugar and is cultivated, and the composition of the substratum of employing is: straw 3g, and wheat bran 1g, Mandels trace element salts solution 0.5mL, nutritive salt 10mL, peptone 0.1g is settled to 100mL with distilled water, and the pH value transfers to 4.8.Contain potassium primary phosphate 0.4% in the wherein said nutritive salt, ammonium sulfate 1.6%, sal epsom 0.1%, distilled water 97.9%, this is meant mass concentration.
In the 3rd step, the Pseudomonas aeruginosa BSZ-07 that will be cultured to logarithmic phase then is inoculated in the straw hydrolyzation system in second step, realizes the online production of rhamnosyl lipid tensio-active agent, to promote the hydrolysis of cellulase.
In described the 3rd step, be specially: Pseudomonas aeruginosa BSZ-07 is dissolved in the 50mL seed culture medium with preparation seed suspension, and keeping density is 10
6Spore/mL.Then, get the seed suspension 5mL of bacterial strain, be inoculated into 100mL fermentation and produce in the tensio-active agent substratum, shaking culture 24h is used to produce bio-surfactant to logarithmic phase in 35 ℃, 200r/min shaking table.The Pseudomonas aeruginosa BSZ-07 seed liquor that 2-6mL is in logarithmic phase is inoculated in the straw hydrolyzation system in second step, and wherein the volume of straw hydrolyzation system is 100mL.Bacterial classification is cultivated 48h in 35 ℃, 200r/min shaking table, be used for the straw hydrolysis.Seed culture medium in this step and fermentation produce thing tensio-active agent substratum can adopt existing substratum.
Described seed culture medium composition is: extractum carnis 3g, NaNO
31g, (NH
4)
2SO
41g, Na
2HPO
41.2g, KH
2PO
41g, MgSO
47H
2O 0.01g, CaCl
20.002g, water 1000mL, pH 6.5-7.0.
The composition that described fermentation produces thing tensio-active agent substratum is: glucose 2g, yeast extract paste 0.1g, NH
4NO
30.5g, machine oil 2mL, KH
2PO
41g, Na
2HPO
41g, MgSO
47H
2O 0.02g, trace element solution 0.5mL, water 1000mL.
With the cellulase hydrolysis of not inoculating Pseudomonas aeruginosa BSZ-07 to comparing in the same old way, the online production of rhamnolipid biological surface activator is added can play significant synergism to cellulose hydrolysis.Because the online production of rhamnolipid can be saved this complicated technology of purification of rhamnolipid, therefore can effectively reduce its production cost, make it become a kind of more potential addition manner.
The present invention is in the process of cellulase producing bacteria-Trichodermareesei ZM4-F3 hydrolysis straw, produce bacterium-Pseudomonas aeruginosa BSZ-07 to wherein directly adding the rhamnolipid biological surface activator that is cultured to logarithmic phase, realized the online production addition manner of rhamnolipid type biological surfactant.Because the addition manner of this kind online production can save this complicated technology of purification of rhamnolipid, therefore can effectively reduce the production cost of rhamnolipid biological surface activator, be applicable to industrial production.In addition, the interpolation of rhamnolipid can effectively improve the hydrolysis efficiency of cellulase.The present invention can effectively reduce bio-surfactant production cost, simplify its production technique, and be applicable to large-scale industrial production.
Described Trichodermareesei ZM4-F3, the preservation information of this bacterial classification can be bought: No. 32, in May, 2006 of Trichodermareesei (Trichoderma reesei), Chinese industrial microbial strains preservation center, Xiaoyun Road, Chaoyang District, Beijing City, numbering 40360 by Chinese industrial microbial strains preservation center (CICC).
Described Pseudomonas aeruginosa BSZ-07, the preservation information of this bacterial classification can be bought: pseudomonas aeruginosa (Pseudomonas aeruginosa), Chinese typical culture collection center, wuchang, wuhan district Luo Jiashan, in May, 2006, numbering AB93066 by China typical culture collection center (CCTCC).
Description of drawings
Fig. 1 be with Pseudomonas aeruginosa BSZ-07 with 4% inoculum size direct inoculation to Trichodermareesei ZM4-F3 hydrocellulose system the time, realize that promptly the addition manner of online production rhamnolipid type biological surfactant and cellulase hydrolysis are to produce the comparison diagram of reducing sugar in the same old way.
Embodiment
Below in conjunction with accompanying drawing embodiments of the invention are elaborated: present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
For the influence of the online production addition manner that confirms the rhamnolipid type biological surfactant to cellulase hydrolysis, utilize 752 type ultraviolet spectrophotometers to measure reducing sugar output, and with the plain enzymic hydrolysis system of individual fibers to contrast in the same old way.
Embodiment 1
Harvested straw is washed 4-5 time with removal impurity and stem knot with tap water, and cut segment to 1-2cm.The straw that cuts places 70 ℃ of baking ovens to be dried to constant weight, at room temperature stores in order to follow-up use.Before enzymic hydrolysis, carry out pre-treatment with shearing good straw with alkali, concrete grammar is: the straw that 10g is sheared places the Erlenmeyer flask of the sodium hydroxide that 40ml 2% is housed, and keeping solid-liquid body mass ratio is 1: 4,85 ℃ of water-bath 1h.Be washed till neutrality with distilled water, place 70 ℃ of baking ovens to be dried to constant weight.
Pretreated straw is produced sugar with Trichodermareesei ZM4-F3 hydrolysis in 30 ℃, 200rpm shaking table to be cultivated.
The Pseudomonas aeruginosa BSZ-07 that will be cultured to logarithmic phase then is inoculated in the straw hydrolyzation system with 2% inoculum size, that is: Pseudomonas aeruginosa BSZ-07 is dissolved in the 50mL seed culture medium with preparation seed suspension, and keeping density is 10
6Spore/mL.Then, get the seed suspension 5mL of bacterial strain, be inoculated into 100mL fermentation and produce in the tensio-active agent substratum, shaking culture 24h is to logarithmic phase, in order to produce bio-surfactant in 35 ℃, 200r/min shaking table.The Pseudomonas aeruginosa BSZ-07 seed liquor that 2mL is in logarithmic phase is inoculated in the straw hydrolyzation system, and wherein the volume of straw hydrolyzation system is 100mL.Bacterial classification is cultivated 48h in 35 ℃, 200r/min shaking table, be used for the straw hydrolysis.
In the present embodiment, described Trichodermareesei ZM4-F3 hydrolysis is produced sugar and is cultivated, and the composition of the substratum of employing is: straw 3g, wheat bran 1g, Mandels trace element salts solution 0.5mL, nutritive salt (potassium primary phosphate 0.4%, ammonium sulfate 1.6%, sal epsom 0.1%, percentage composition herein refers to mass concentration) 10mL, peptone 0.1g is settled to 100mL with distilled water, and the pH value transfers to 4.8.
Described seed culture medium composition is: extractum carnis 3g, NaNO
31g, (NH
4)
2SO
41g, Na
2HPO
41.2g, KH
2PO
41g, MgSO
47H
2O 0.01g, CaCl
20.002g, water 1000mL, pH 6.5-7.0.
The composition that the tensio-active agent substratum is produced in described fermentation is: glucose 2g, yeast extract paste 0.1g, NH
4NO
30.5g, machine oil 2mL, KH
2PO
41g, Na
2HPO
41g, MgSO
47H
2O 0.02g, trace element solution 0.5mL, water 1000mL.
Present embodiment adopts the online production of Trichodermareesei ZM4-F3 and Pseudomonas aeruginosa BSZ-07 realization rhamnolipid biological surface activator to add, to promote the hydrolysis of cellulase.And with the plain enzymic hydrolysis system of individual fibers to contrast in the same old way.
Discover that the online production addition manner of rhamnolipid is about to Pseudomonas aeruginosa BSZ-07 and, promptly increases sharply from its reducing sugar output of 48h to Trichodermareesei ZM4-F3 hydrocellulose system the time with 2% inoculum size direct inoculation.To in the same old way, 96h is that the optimum of Trichodermareesei ZM4-F3 produces the sugar time, and it produced reducing sugar amount and reached maximum value this moment, was 2.231g/l.And with Pseudomonas aeruginosa BSZ-07 with 2% inoculum size direct inoculation to Trichodermareesei ZM4-F3 hydrocellulose system the time, just can make the reducing sugar amount reach 2.277g/l when producing sugared 84h.Therefore, the online production addition manner of this kind rhamnolipid can not only increase the output of reducing sugar, can also obviously shorten optimum producing the sugar time.
Embodiment 2
Harvested straw is washed 4-5 time with removal impurity and stem knot with tap water, and cut segment to 1-2cm.The straw that cuts places 70 ℃ of baking ovens to be dried to constant weight, at room temperature stores in order to follow-up use.Before enzymic hydrolysis, carry out pre-treatment with shearing good straw with alkali, concrete grammar is: the straw that 10g is sheared places the Erlenmeyer flask of the sodium hydroxide that 40ml 2% is housed, and keeping solid-liquid body mass ratio is 1: 4,85 ℃ of water-bath 1h.Be washed till neutrality with distilled water, place 70 ℃ of baking ovens to be dried to constant weight.
Pretreated straw is produced sugar with Trichodermareesei ZM4-F3 hydrolysis in 30 ℃, 200rpm shaking table to be cultivated.
The Pseudomonas aeruginosa BSZ-07 that will be cultured to logarithmic phase then is inoculated in the straw hydrolyzation system with 4% inoculum size: Pseudomonas aeruginosa BSZ-07 is dissolved in the 50mL seed culture medium with preparation seed suspension, and keeping density is 10
6Spore/mL.Then, get the seed suspension 5mL of bacterial strain, be inoculated into 100mL fermentation and produce in the tensio-active agent substratum, shaking culture 24h is to logarithmic phase, in order to produce bio-surfactant in 35 ℃, 200r/min shaking table.The Pseudomonas aeruginosa BSZ-07 seed liquor that 4mL is in logarithmic phase is inoculated in the straw hydrolyzation system, and wherein the volume of straw hydrolyzation system is 100mL.Bacterial classification is cultivated 48h in 35 ℃, 200r/min shaking table, be used for the straw hydrolysis.
Wherein, described Trichodermareesei ZM4-F3 hydrolysis is produced sugar and is cultivated, and the composition of the substratum of employing is: straw 3g, wheat bran 1g, Mandels trace element salts solution 0.5mL, nutritive salt (potassium primary phosphate 0.4%, ammonium sulfate 1.6%, sal epsom 0.1%, percentage composition herein refers to mass concentration) 10mL, peptone 0.1g is settled to 100mL with distilled water, and the pH value transfers to 4.8.
Described seed culture medium composition is: extractum carnis 3g, NaNO
31g, (NH
4)
2SO
41g, Na
2HPO
41.2g, KH
2PO
41g, MgSO
47H
2O 0.01g, CaCl
20.002g, water 1000mL, pH 6.5-7.0.
The composition that the tensio-active agent substratum is produced in described fermentation is: glucose 2g, yeast extract paste 0.1g, NH
4NO
30.5g, machine oil 2mL, KH
2PO
41g, Na
2HPO
41g, MgSO
47H
2O 0.02g, trace element solution 0.5mL, water 1000mL.
Present embodiment adopts the online production of Trichodermareesei ZM4-F3 and Pseudomonas aeruginosa BSZ-07 realization rhamnolipid biological surface activator to add, to promote the hydrolysis of cellulase.And with the plain enzymic hydrolysis system of individual fibers to contrast in the same old way.
Discover that the online production addition manner of rhamnolipid is about to Pseudomonas aeruginosa BSZ-07 and, promptly increases sharply from its reducing sugar output of 48h to Trichodermareesei ZM4-F3 hydrocellulose system the time with 4% inoculum size direct inoculation.To in the same old way, 96h is that the optimum of Trichodermareesei ZM4-F3 produces the sugar time, and it produced reducing sugar amount and reached maximum value this moment, was 2.231g/l.And with Pseudomonas aeruginosa BSZ-07 with 4% inoculum size direct inoculation to Trichodermareesei ZM4-F3 hydrocellulose system the time, just can make the reducing sugar amount reach 2.571g/l when producing sugared 84h.Therefore, the online production addition manner of this kind rhamnolipid can not only increase the output of reducing sugar, can also obviously shorten optimum producing the sugar time.
Embodiment 3
Harvested straw is washed 4-5 time with removal impurity and stem knot with tap water, and cut segment to 1-2cm.The straw that cuts places 70 ℃ of baking ovens to be dried to constant weight, at room temperature stores in order to follow-up use.Before enzymic hydrolysis, carry out pre-treatment with shearing good straw with alkali, concrete grammar is: the straw that 10g is sheared places the Erlenmeyer flask of the sodium hydroxide that 40ml 2% is housed, and keeping solid-liquid body mass ratio is 1: 4,85 ℃ of water-bath 1h.Be washed till neutrality with distilled water, place 70 ℃ of baking ovens to be dried to constant weight.
Pretreated straw is produced sugar with Trichodermareesei ZM4-F3 hydrolysis in 30 ℃, 200rpm shaking table to be cultivated.
The Pseudomonas aeruginosa BSZ-07 that will be cultured to logarithmic phase then is inoculated in the straw hydrolyzation system with 6% inoculum size: Pseudomonas aeruginosa BSZ-07 is dissolved in the 50mL seed culture medium with preparation seed suspension, and keeping density is 10
6Spore/mL.Then, get the seed suspension 5mL of bacterial strain, be inoculated into 100mL fermentation and produce in the tensio-active agent substratum, shaking culture 24h is to logarithmic phase, in order to produce bio-surfactant in 35 ℃, 200r/min shaking table.The Pseudomonas aeruginosa BSZ-07 seed liquor that 6mL is in logarithmic phase is inoculated in the straw hydrolyzation system, and wherein the volume of straw hydrolyzation system is 100mL.Bacterial classification is cultivated 48h in 35 ℃, 200r/min shaking table, be used for the straw hydrolysis.
Wherein, described Trichodermareesei ZM4-F3 hydrolysis is produced sugar and is cultivated, and the composition of the substratum of employing is: straw 3g, wheat bran 1g, Mandels trace element salts solution 0.5mL, nutritive salt (potassium primary phosphate 0.4%, ammonium sulfate 1.6%, sal epsom 0.1%, percentage composition herein refers to mass concentration) 10mL, peptone 0.1g is settled to 100mL with distilled water, and the pH value transfers to 4.8.
Described seed culture medium composition is: extractum carnis 3g, NaNO
31g, (NH
4)
2SO
41g, Na
2HPO
41.2g, KH
2PO
41g, MgSO
47H
2O 0.01g, CaCl
20.002g, water 1000mL, pH 6.5-7.0.
The composition that the tensio-active agent substratum is produced in described fermentation is: glucose 2g, yeast extract paste 0.1g, NH
4NO
30.5g, machine oil 2mL, KH
2PO
41g, Na
2HPO
41g, MgSO
47H
2O 0.02g, trace element solution 0.5mL, water 1000mL.
Present embodiment adopts the online production of Trichodermareesei ZM4-F3 and Pseudomonas aeruginosa BSZ-07 realization rhamnolipid biological surface activator to add, to promote the hydrolysis of cellulase.And with the plain enzymic hydrolysis system of individual fibers to contrast in the same old way.
Discover that the online production addition manner of rhamnolipid is about to Pseudomonas aeruginosa BSZ-07 and, promptly increases sharply from its reducing sugar output of 48h to Trichodermareesei ZM4-F3 hydrocellulose system the time with 6% inoculum size direct inoculation.To in the same old way, 96h is that the optimum of Trichodermareesei ZM4-F3 produces the sugar time, and it produced reducing sugar amount and reached maximum value this moment, was 2.231g/l.And with Pseudomonas aeruginosa BSZ-07 with 6% inoculum size direct inoculation to Trichodermareesei ZM4-F3 hydrocellulose system the time, just can make the reducing sugar amount reach 2.432g/l when producing sugared 84h.Therefore, the online production addition manner of this kind rhamnolipid can not only increase the output of reducing sugar, can also obviously shorten optimum producing the sugar time.
As can be seen from Figure 1, when having only the independent hydrolysis straw of Trichodermareesei ZM4-F3 to comparing in the same old way, Pseudomonas aeruginosa BSZ-07 and Trichodermareesei ZM4-F3 cohydrolysis sample increase sharply and surpass value in the same old way from the reducing sugar output of 48h.To in the same old way, 96h is that the optimum of Trichodermareesei ZM4-F3 produces the sugar time, and it produced reducing sugar amount and reached maximum value this moment, was 2.231g/L.And in cohydrolysis (being online production) sample, just can make the reducing sugar amount reach 2.571g/L when producing sugared 84h.Therefore result of study shows that the interpolation of Pseudomonas aeruginosa BSZ-07 in the rice straw degradation process can not only increase the output of reducing sugar, can also obviously shorten the rice straw degradation time.
Claims (9)
1, the online production method of rhamnolipid biological surface activator in a kind of cellulase hydrolysis is characterized in that, comprises the steps:
The first step is cleaned harvested straw, cut off, and is dried to constant weight, before cellulase hydrolysis, the straw that cuts is handled with alkali;
Second goes on foot, and the straw after handling is hydrolyzed with Trichodermareesei ZM4-F3 produces the sugar cultivation, in order to the generation reducing sugar;
In the 3rd step, the Pseudomonas aeruginosa BSZ-07 that will be cultured to logarithmic phase then is inoculated in the straw hydrolyzation system in second step, realizes the online production of rhamnosyl lipid tensio-active agent, promotes the hydrolysis of cellulase.
2, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 1, it is characterized in that, described harvested straw is clean, cut-out, be dried to constant weight, be meant: harvested straw is washed 4-5 all over tying to remove impurity and stem with tap water, and cut segment to 1-2cm, the straw that cuts places 70 ℃ of baking ovens to be dried to constant weight, at room temperature stores.
3, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 1, it is characterized in that, the described straw well of will shearing is handled with alkali, be meant: the straw that 10g is sheared places the sodium hydroxide of 40ml 2% volumetric concentration, keeping solid-liquid body mass ratio is 1: 4,85 ℃ of water-bath 1h are washed till neutrality with distilled water, place 70 ℃ of baking ovens to be dried to constant weight.
4, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 1 is characterized in that, described being hydrolyzed with Trichodermareesei ZM4-F3 produced sugar and cultivated, and is to carry out in 30 ℃, 200rpm shaking table.
5, according to the online production method of rhamnolipid biological surface activator in claim 1 or the 4 described cellulase hydrolysiss, it is characterized in that, described being hydrolyzed with Trichodermareesei ZM4-F3 produced the sugar cultivation, the composition of the substratum that adopts is: straw 3g, wheat bran 1g, Mandels trace element salts solution 0.5mL, nutritive salt 10mL, peptone 0.1g is settled to 100mL with distilled water, and the pH value transfers to 4.8.
6, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 5, it is characterized in that constituent mass concentration in the described nutritive salt: potassium primary phosphate 0.4%, ammonium sulfate 1.6%, sal epsom 0.1%, distilled water 97.9%.
7, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 1, it is characterized in that, in described the 3rd step, be specially: Pseudomonas aeruginosa BSZ-07 is dissolved in the 50mL seed culture medium with preparation seed suspension, and keeping density is 10
6Spore/mL; Then, get the seed suspension 5mL of bacterial strain, be inoculated into 100mL fermentation and produce in the tensio-active agent substratum, shaking culture 24h is to logarithmic phase in 35 ℃, 200r/min shaking table; The Pseudomonas aeruginosa BSZ-07 seed liquor that 2-6mL is in logarithmic phase is inoculated in the straw hydrolyzation system, and wherein the volume of straw hydrolyzation system is 100mL, and bacterial classification is cultivated 48h in 35 ℃, 200r/min shaking table, be used for the straw hydrolysis.
8, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 7 is characterized in that, described seed culture medium composition is: extractum carnis 3g, NaNO
31g, (NH
4)
2SO
41g, Na
2HPO
41.2g, KH
2PO
41g, MgSO
47H
2O 0.01g, CaCl
20.002g, water 1000mL, pH 6.5-7.0.
9, the online production method of rhamnolipid biological surface activator in the cellulase hydrolysis according to claim 7 is characterized in that, the composition that described fermentation produces thing tensio-active agent substratum is: glucose 2g, yeast extract paste 0.1g, NH
4NO
30.5g, machine oil 2mL, KH
2PO
41g, Na
2HPO
41g, MgSO
47H
2O0.02g, trace element solution 0.5mL, water 1000mL.
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| CN101898100A (en) * | 2010-04-13 | 2010-12-01 | 东北农业大学 | Biosurfactant as well as preparation method and application thereof |
| CN101898907A (en) * | 2010-04-13 | 2010-12-01 | 东北农业大学 | Compound microbial preparation and application thereof in microbial degradation of crop straw |
| CN101898907B (en) * | 2010-04-13 | 2013-07-10 | 东北农业大学 | Compound microbial preparation and application thereof in microbial degradation of crop straw |
| CN101898100B (en) * | 2010-04-13 | 2013-10-23 | 东北农业大学 | Biosurfactant as well as preparation method and application thereof |
| CN103088101A (en) * | 2013-01-18 | 2013-05-08 | 北京林业大学 | Method for extracting natural saponin and biosurfactant composite product |
| CN103088101B (en) * | 2013-01-18 | 2014-12-10 | 北京林业大学 | Method for extracting natural saponin and biosurfactant composite product |
| CN103305447A (en) * | 2013-07-04 | 2013-09-18 | 江苏德鑫环保科技有限公司 | Organic waste degradation bacterium and preparation method thereof |
| CN103305447B (en) * | 2013-07-04 | 2015-08-05 | 江苏德鑫环保科技有限公司 | A kind of Organic waste degradation bacterium and method for making thereof |
| US9884883B2 (en) | 2015-01-12 | 2018-02-06 | Logos Technologies, Llc | Production of rhamnolipid compositions |
| US10829507B2 (en) | 2017-02-06 | 2020-11-10 | Stepan Company | Decolorization of concentrated rhamnolipid composition |
| CN111793665A (en) * | 2019-04-09 | 2020-10-20 | 中国科学院微生物研究所 | Production method of biological surface active functional protein and culture medium used by same |
| CN111793665B (en) * | 2019-04-09 | 2022-03-15 | 中国科学院微生物研究所 | Production method of biological surface active functional protein and culture medium used by same |
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