CN104911164A - Method for improving cellulase use efficiency, and apparatus thereof - Google Patents
Method for improving cellulase use efficiency, and apparatus thereof Download PDFInfo
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- CN104911164A CN104911164A CN201510234945.4A CN201510234945A CN104911164A CN 104911164 A CN104911164 A CN 104911164A CN 201510234945 A CN201510234945 A CN 201510234945A CN 104911164 A CN104911164 A CN 104911164A
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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Abstract
The invention relates to a method for improving the cellulase use efficiency, and an apparatus thereof. The method comprises the following steps: 1, extracting hemicellulose by using diluted sulfuric acid, immersing in an alkali to remove lignin, and processing a cellulose raw material; and 2, hydrolyzing the processed cellulose raw material by cellulase in an enzymatic hydrolysis reactor. The method and the apparatus avoid adsorption of lignin to cellulase and increase the hemicellulose utilization rate, and the enzymatic hydrolysis reactor is not provided with a stirring device to solve a problem of enzyme inactivation caused by excessive stirring of traditional enzymatic hydrolysis reactors; a traditional intermittence technology is changed to a continuous technology, so the product inhibition problem in the process is solved; and the cellulase use amount in the method and the apparatus in the invention is obviously lower than that of traditional technologies.
Description
Technical field
This patent relates to and uses cellulase when carrying out enzymolysis to Mierocrystalline cellulose, improves service efficiency, reduces the method and apparatus of usage quantity.
Background technology
Cellulose hydrolysis technique means mainly comprises dilute sulphuric acid hydrolysis and cellulase hydrolysis at present, wherein the efficiency of enzymic hydrolysis and the selectivity of final product and purity higher, but the cost of the high cellulase of enzymic hydrolysis is the difficulty being difficult at present overcome.
Under the state of the art of current cellulase, cause the too high reason of cellulase use cost have following some:
1, relevant with the formation structure of plant itself.The cellulose crystallity of some plants is high, compact structure, although through various means pre-treatment, is still difficult to destroy structure, contacts to make cellulase with Mierocrystalline cellulose.
2, after pre-treatment, a large amount of xylogen in vegetable fibre, is still had to remain.Xylogen can adsorptive cellulose enzyme, makes to reduce with the enzyme amount of fibrin reaction.
3, common hydrolysis flow process is all intermittent type, degrade gradually along with cellulosic, its content is fewer and feweri, and the probability that therefore cellulase contacts with Mierocrystalline cellulose is also reducing gradually, and now still having part enzyme to have high vigor in system, this is also one of factor reducing cellulase service efficiency; The phenomenon of Product inhibiton can be produced in the enzyme digestion reaction process of interval, enzymolysis efficiency can be caused equally to reduce.
4, common enzymolysis reactor is all equipped with agitator, is uniformly mixed enzymic hydrolysis system, promotes cellulase and cellulosic combination.But find in research, cellulase is very responsive for the shearing of stirring, and excessive stirring can cause the inactivation of enzyme
At present, in disclosed documents and materials, the usage quantity of cellulase is generally not less than 10% of Mierocrystalline cellulose total amount, and in hydrolyzed solution, total reducing sugars content is not more than 8%.
Summary of the invention
The object of the invention is to for the problems referred to above, provide a kind of method and apparatus improving cellulase service efficiency, the chance that increased fiber element enzyme contacts with Mierocrystalline cellulose, continuous processing is changed into by traditional batch technology, solve the Product inhibiton problem in process, reduce the usage quantity of cellulase.
The present invention is achieved by the following technical programs.
Improve a method for cellulase service efficiency, method is as follows:
(1) process of cellulosic material
1. functional quality concentration be 5% ~ 10% dilute sulphuric acid process cellulosic material extract hemicellulose in cellulosic material, the mass ratio of dilute sulphuric acid and cellulosic material is 3:1;
2. the solution after step (1) 1. processes is filtered out solid, solid clear water being washed till pH is 6.5 ~ 7.0; Collect filtrate, now filtrate is the dilute sulphuric acid liquid containing hemicellulose simultaneously;
3. by 2. process through step (1) the solid mass concentration obtained be 15 ~ 20% aqueous sodium hydroxide solution soak more than 3 hours under the condition of 120 degree, to remove xylogen, leach to obtain the cellulosic material handled well; The mass ratio of aqueous sodium hydroxide solution and solid is 10:1;
4. the dilute sulphuric acid liquid containing hemicellulose continues to carry out step (1) to the Mierocrystalline cellulose not carrying out processing and 1. processes, 2. the dilute sulphuric acid liquid of hemicellulose must be contained again through step (1), can Reusability 4 ~ 6 times, to the final diluted acid liquid functional quality concentration containing the hemicellulose aqueous sodium hydroxide solution that is 2 ~ 5% be neutralized to pH be 4 ~ 4.5 dilute acid hydrolysis liquid;
(2) hydrolysis of cellulase
1. add in enzymolysis reactor by the cellulosic material handled well, compressed by cellulosic material, the liquid of extrusion is discharged Posterior circle and is returned in enzymolysis reactor;
2. repeating step (2) 1., compresses cellulosic material and make it fill up container, keep pressure to be 1.5 ~ 3kg/cm2 after compressing;
3. use water and dilute acid hydrolysis liquid Novi to be believed cellulase Carezyme 4500L is mixed with the cellulase aqueous solution of 0.1 ~ 0.5%, the dosage of Novi letter cellulase Carezyme 4500L is 3 ~ 5% of content of cellulose in the cellulosic material handled well; Cellulase aqueous solution volume total amount is 10 ~ 15 times of content of cellulose in the cellulosic material handled well;
4. the cellulase aqueous solution is entered enzymolysis reactor, the volumetric flow rate of the cellulase aqueous solution is 1/48 of reaction vessel volume, and inlet pressure is greater than 3kg/cm2, when outlet has liquid to flow out, use protein assay to detect protein content, use liquid chromatography inspection total reducing sugars content; When discovery protein content continues to increase, and obtain cellulosic hydrolysate when total reducing sugars continues minimizing more than 10%, now stop the cellulase aqueous solution to enter, stop the outflow of outlet liquid, continue repeating step (2) 1. 2. 4..
A kind of device improving cellulase service efficiency, described device is enzymolysis reactor, described enzymolysis reactor comprises solution tank body and hydraulic lift, tank body be provided with can cut off charging opening, the outlet of cellulosic hydrolysate and raffinate mouth, tank body lower part is provided with the import of the cellulase aqueous solution, the hydraulic lift of tank body top mounting strap back pressure monitoring, hydraulic lift top is connected with electric motor, hydraulic lift bottom is connected with piston pressing plate, drives hydraulic lift control piston pressing plate to move up and down in tank body by electric motor.
Beneficial effect of the present invention is:
Using certain density aqueous sodium hydroxide solution to carry out pre-treatment to cellulosic material can very effective removal xylogen, avoids xylogen to the absorption of cellulase; But because hemicellulose and xylogen combine more tight, the loss of the hemicellulose of a great deal of can be caused, first use certain density sulfuric acid first to be extracted by the hemicellulose in cellulosic material for this reason, avoid in utilization ratio delignification being reduced thoroughly hemicellulose simultaneously.
In the present invention, enzymolysis reactor eliminates agitator, solve traditional enzymolysis reactor causes the inactivation of enzyme problem due to excessive stirring, and technique is changed into continuous processing by traditional batch technology by this enzymolysis reactor, solve the Product inhibiton problem in process, the accumulation that can not produce substrate presses down, and increases cellulase and cellulosic contact area and time simultaneously.
Use method and apparatus cellulase hydrolysis cellulosic material of the present invention, the usage quantity of cellulase is lower than 5% of content of cellulose.
Accompanying drawing explanation
Fig. 1 is the structure iron of enzymolysis reactor of the present invention.
Embodiment
Below by way of specific embodiment, the specific embodiment of the present invention is described in further detail.
Cellulosic material of the present invention is commercially available bagasse, dries the bagasse obtaining for 4 ~ 6 hours doing at 105 DEG C, and detection fibers element and hemicellulose level are 72%.
Embodiment 1
(1) process of bagasse
1. take the dry bagasse of 10kg, use 30L mass concentration be 5% dilute sulphuric acid process cellulosic material extract hemicellulose in cellulosic material;
2. the solution after step (1) 1. processes is filtered out solid, solid clear water being washed till pH is 6.5 ~ 7.0; Collect filtrate, now filtrate is the dilute sulphuric acid liquid containing hemicellulose simultaneously;
3. by 2. process through step (1) the solid mass concentration obtained be 15% aqueous sodium hydroxide solution soak 5 hours under the condition of 120 degree, to remove xylogen, leach to obtain the bagasse handled well, the mass ratio of aqueous sodium hydroxide solution and solid is 10:1;
4. the dilute sulphuric acid liquid containing hemicellulose continues to carry out step (1) to the Mierocrystalline cellulose not carrying out processing and 1. processes, 2. the dilute sulphuric acid liquid of hemicellulose must be contained again through step (1), can Reusability 4 ~ 6 times, to the diluted acid liquid containing hemicellulose that 30L is final use 15L mass concentration be 5% aqueous sodium hydroxide solution be neutralized to pH be 4.5 dilute acid hydrolysis liquid;
(2) hydrolysis of cellulase
1. behind starter motor 1 jigger lifting piston pressing plate 3 to reactor tank body 8 top, the bagasse handled well is added in the enzymolysis reactor of 2L by charging opening 4, actuating motor 1 drives piston pressing plate 3 to be compressed by bagasse, and the liquid of extrusion is discharged Posterior circle from raffinate mouth 7 and returned feed storage tank;
2. repeating step (2) 1., compresses cellulosic material and make it fill up container, keeps the back pressure of hydraulic lift 2 to be designated as 2kg/cm2 after compressing.
3. use 27L tap water and dilute acid hydrolysis liquid Novi to be believed cellulase Carezyme 4500L is mixed with the cellulase aqueous solution of 0.3%, the dosage of Novi letter cellulase Carezyme 4500L is 0.216kg; Cellulase aqueous solution volume total amount is 72L;
4. the cellulase aqueous solution is entered enzymolysis reactor by the aqueous solution import 6 of cellulase, the volumetric flow rate of the cellulase aqueous solution is 42ml/h, inlet pressure is greater than 3kg/cm2, when hydrolyzed solution outlet 5 has liquid to flow out, using protein assay to detect protein content is 0.001%, uses liquid chromatography inspection total reducing sugars content substantially to maintain 30g/L; When discovery protein content continues to increase, and obtain cellulosic hydrolysate when total reducing sugars continues to be reduced to below 27g/L, now stop the cellulase aqueous solution to enter, stop the outflow of hydrolyzed solution outlet liquid, continue repeating step (2) 1. 2. 4..
The actual cellulosic hydrolysate collected is 69L, and reducing sugar total content is 9.5%.Actual enzyme usage quantity is lower than 5% of content of cellulose.
As can be seen from the above-described embodiment, adopt the present invention to improve the method and apparatus of cellulase service efficiency, the usage quantity of cellulase is starkly lower than traditional method, reduces production cost.
Here description of the invention and application is illustrative, not wants by scope restriction of the present invention in the above-described embodiments, and therefore, the present invention is not by the restriction of the present embodiment, and the technical scheme that any employing equivalence replacement obtains is all in the scope of protection of the invention.
Claims (2)
1. improve a method for cellulase service efficiency, it is characterized by, method is as follows:
(1) process of cellulosic material
1. functional quality concentration be 5% ~ 10% dilute sulphuric acid process cellulosic material extract hemicellulose in cellulosic material, the mass ratio of dilute sulphuric acid and cellulosic material is 3:1;
2. the solution after step (1) 1. processes is filtered out solid, solid clear water being washed till pH is 6.5 ~ 7.0; Collect filtrate, now filtrate is the dilute sulphuric acid liquid containing hemicellulose simultaneously;
3. by 2. process through step (1) the solid mass concentration obtained be 15 ~ 20% aqueous sodium hydroxide solution soak more than 3 hours under the condition of 120 degree, to remove xylogen, leach to obtain the cellulosic material handled well; The mass ratio of aqueous sodium hydroxide solution and solid is 10:1;
4. the dilute sulphuric acid liquid containing hemicellulose continues to carry out step (1) to the Mierocrystalline cellulose not carrying out processing and 1. processes, 2. the dilute sulphuric acid liquid of hemicellulose must be contained again through step (1), can Reusability 4 ~ 6 times, to the final diluted acid liquid functional quality concentration containing the hemicellulose aqueous sodium hydroxide solution that is 2 ~ 5% be neutralized to pH be 4 ~ 4.5 dilute acid hydrolysis liquid;
(2) hydrolysis of cellulase
1. add in enzymolysis reactor by the cellulosic material handled well, compressed by cellulosic material, the liquid of extrusion is discharged Posterior circle and is returned in enzymolysis reactor;
2. repeating step (2) 1., compresses cellulosic material and make it fill up container, keep pressure to be 1.5 ~ 3kg/cm2 after compressing; 3. use water and dilute acid hydrolysis liquid Novi to be believed cellulase Carezyme 4500L is mixed with the cellulase aqueous solution of 0.1 ~ 0.5%, the dosage of Novi letter cellulase Carezyme 4500L is 3 ~ 5% of content of cellulose in the cellulosic material handled well; Cellulase aqueous solution volume total amount is 10 ~ 15 times of content of cellulose in the cellulosic material handled well;
4. the cellulase aqueous solution is entered enzymolysis reactor, the volumetric flow rate of the cellulase aqueous solution is 1/48 of reaction vessel volume, and inlet pressure is greater than 3kg/cm2, when outlet has liquid to flow out, use protein assay to detect protein content, use liquid chromatography inspection total reducing sugars content; When discovery protein content continues to increase, and obtain cellulosic hydrolysate when total reducing sugars continues minimizing more than 10%, now stop the cellulase aqueous solution to enter, stop the outflow of outlet liquid, continue repeating step (2) 1. 2. 4..
2. one kind is improved the device of cellulase service efficiency, it is characterized by, described device is enzymolysis reactor, described enzymolysis reactor comprises solution tank body and hydraulic lift, tank body be provided with can cut off charging opening, the outlet of cellulosic hydrolysate and raffinate mouth, tank body lower part is provided with the import of the cellulase aqueous solution, the hydraulic lift of tank body top mounting strap back pressure monitoring, hydraulic lift top is connected with electric motor, hydraulic lift bottom is connected with piston pressing plate, drives hydraulic lift control piston pressing plate to move up and down in tank body by electric motor.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105567567A (en) * | 2016-02-03 | 2016-05-11 | 程雪娇 | Bagasse culture medium and preparing method thereof |
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