CN104846032A - Method for preparing cellulose hydrolyzed sugar by using cellulase - Google Patents
Method for preparing cellulose hydrolyzed sugar by using cellulase Download PDFInfo
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- CN104846032A CN104846032A CN201510162833.2A CN201510162833A CN104846032A CN 104846032 A CN104846032 A CN 104846032A CN 201510162833 A CN201510162833 A CN 201510162833A CN 104846032 A CN104846032 A CN 104846032A
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Abstract
The invention relates to a method for preparing cellulose hydrolyzed sugar by using cellulase. Lactalbumin and other active matters are added before adding cellulase to fill the active site of lignin in advance, so the addition amount of cellulase is greatly reduced. No surfactants are contained in a system, and a cellulose hydrolyzed sugar solution obtained after treatment in above mode can be used in a biological fermentation system, so the treatment cost is reduced, and the output of the cellulose hydrolyzed sugar is improved.
Description
Technical field
The present invention relates to and use cellulase to be hydrolyzed for pretreated Mierocrystalline cellulose to obtain the method for cellulose hydrolysis sugar.
Background technology
In recent years, along with the exhaustion day by day of Nonrenewable resources, the research emphasis of academia in new forms of energy starts to renewable resources, such as vegetable fibre, non-grain crop starch etc.And use cellulase hydrolysis to produce cellulose hydrolysis sugar, and rear application is carried out to it become study hotspot gradually
Current use cellulase hydrolysis prepare hydrolysis sugar carry out fermentation main difficult point have two:
1, current, the bacterial classification of the maturation that fermentation industry uses mostly cannot metabolism five-carbon sugar, and containing the five-carbon sugar of at least 20% in cellulosic hydrolysate, therefore greatly reduces the service efficiency of cellulosic hydrolysate in fermentation industry.
2, when Mierocrystalline cellulose carries out enzymolysis, the cellulase cost of use is very high.Even if carried out pre-treatment to Mierocrystalline cellulose, such as, acidolysis, alkaline hydrolysis, the means such as steam explosion, the xylogen that all cannot be contained all is removed, and xylogen is very large for cellulosic adsorption, the cellulase really contacted with Mierocrystalline cellulose in system is reduced greatly.
For solving the problem, document is had to use the tensio-active agents such as tween20, tween80 to be closed in residual lignin surface adsorption active centre, the consumption of the enzyme used when greatly can reduce cellulose hydrolysis.Although tensio-active agent usage quantity is higher, it is less expensive, and therefore processing cost increases seldom.
But, if by when doing aftertreatment after using the cellulose hydrolysis sugar soln of tensio-active agent to ferment, larger trouble can be run into.Aftertreatment technology many employings micro-filtration of current fermentation, ultrafiltration, the membrane process such as nanofiltration are purified, and the internal structure mechanism of film determines that its Surfactant is more responsive, and tensio-active agent can cause flux depression, even the situations such as blocking occur.
Summary of the invention
The object of the invention is to overcome defect of the prior art, a kind of method using cellulase to prepare cellulose hydrolysis sugar is provided, whey-protein isoreactivity thing was added before interpolation cellulase, the effect being pre-charged with xylogen active centre can be played, adding of cellulase can be reduced in a large number, not containing tensio-active agent in whole system, cellulose hydrolysis liquid glucose through aforesaid way process can be used in biological fermentation system, reduce processing cost, improve the output of cellulose hydrolysis sugar.
The present invention is achieved by the following technical programs.
Use cellulase to prepare a method for cellulose hydrolysis sugar, its preparation method is as follows:
(1) pre-treatment of cellulosic material:
1. cellulosic material is taken by the mass ratio of 1:18 ~ 1:22 and volumetric molar concentration is the aqueous sodium hydroxide solution of 1%;
2. cellulosic material is soaked 1-3 hour in the aqueous sodium hydroxide solution of 110-130 DEG C;
3. soak after terminating, cellulosic material is taken out, use washed with de-ionized water cellulosic material until PH is 6.8-7.2, then add after the pH value of cellulosic material is adjusted to 4.0-4.5 by organic acid and obtain the elementary material of Mierocrystalline cellulose;
4. in the elementary material of Mierocrystalline cellulose, actives is added, hybrid reaction more than 2 hours at the temperature of 38-40 DEG C, Mierocrystalline cellulose is finally expected, the quality of described actives is the 0.01%-0.05% of cellulosic material quality, and described actives is that molecular weight is at 8000 ~ 110000 daltonian egg white mixtures;
(2) added by cellulase during Mierocrystalline cellulose is finally expected and be hydrolyzed, hydrolysis time is 45-50 hour, can obtain cellulose hydrolysis sugar, and the quality of described cellulase is 5% of cellulosic material quality.
A kind of above-mentioned method using cellulase to prepare cellulose hydrolysis sugar, wherein, described cellulosic material comprises maize straw, paddy stalk, bagasse, Herba Eichhorniae.
A kind of above-mentioned method using cellulase to prepare cellulose hydrolysis sugar, wherein, the quality of described actives is the 0.3%-0.5% of cellulosic material quality.
A kind of above-mentioned method using cellulase to prepare cellulose hydrolysis sugar, wherein, described egg white mixture is composed of the following components by mass percentage:
Alpha-lactalbumin 0.1-1%
Bovine serum albumin 0.1-1%
Peptone 98-99.8%.
The mechanism of action of cellulase in cellulose hydrolysis is substantially distinct, since xylogen belongs to ineffective adsorption for cellulase absorption for the whole process of cellulase hydrolysis, if the adsorption activity center of the material so using some and cellulase similar first filling timber quality, add the ratio that cellulase just can reduce ineffective adsorption again, thus be equivalent to improve the ratio that cellulase contacts with Mierocrystalline cellulose.
Cellulase is some protein matters mainly, main composition structure is different from other protein varieties, but it is identical for can playing adsorbing functional group, therefore, must screen some roughly the same with Cellulase Molecules amount, the close proteic substance of character can be adsorbed by alternative fiber element enzyme.
Find in research, the molecular weight of cellulase constitute, from 8000 dalton to 110,000 dalton, through screening and mixture, uses actives of the present invention can the formation of mimic cellulose enzyme.
The main protein component of actives of the present invention has alpha-lactalbumin, peptone, bovine serum albumin, and the scope of overall molecule amount is also from 8000 to 11 ten thousand dalton.
Experiment proves, actives of the present invention was added before interpolation cellulase, the effect being pre-charged with xylogen active centre can be played, adding of cellulase can be reduced in a large number, not containing tensio-active agent in whole system, cellulose hydrolysis liquid glucose through aforesaid way process can be used in biological fermentation system, reduces processing cost, improves the output of cellulose hydrolysis sugar.
Embodiment
Below by way of specific embodiment, the specific embodiment of the present invention is described in further detail.
Embodiment one
Use cellulase to prepare a method for cellulose hydrolysis sugar, its preparation method is as follows:
(1) pre-treatment of raw material:
1. after taking bagasse oven dry, raw material 50g and volumetric molar concentration are the aqueous sodium hydroxide solution 1000g of 1%;
2. bagasse raw material is soaked 2 hours in the aqueous sodium hydroxide solution of 120 DEG C;
3. soak after terminating, bagasse raw material is taken out, use washed with de-ionized water bagasse raw material until PH is 7.0, then add after the pH value of bagasse raw material is adjusted to 4.5 by citric acid and obtain the elementary material of bagasse;
4. alpha-lactalbumin 0.5% is got by mass percentage, bovine serum albumin 0.7%, peptone ratio is 98.8% be mixed with into protein-active thing, adds protein-active thing 0.025g in the elementary material of bagasse, hybrid reaction 2.5 hours at the temperature of 39 DEG C, bagasse is finally expected;
(2) added during bagasse is finally expected by cellulase 2.5g and be hydrolyzed, hydrolysis time is 48 hours, can obtain cellulose hydrolysis sugar.
Embodiment two
Use cellulase to prepare a method for cellulose hydrolysis sugar, its preparation method is as follows:
(1) pre-treatment of raw material:
1. after taking maize straw oven dry, raw material 50g and volumetric molar concentration are the aqueous sodium hydroxide solution 1050g of 1%;
2. corn stalk raw material is soaked 2 hours in the aqueous sodium hydroxide solution of 120 DEG C;
3. soak after terminating, corn stalk raw material is taken out, use washed with de-ionized water corn stalk raw material until PH is 7.0, then add after the pH value of corn stalk raw material is adjusted to 4.2 by citric acid and obtain the elementary material of maize straw;
4. alpha-lactalbumin 0.5% is got by mass percentage, bovine serum albumin 1%, peptone ratio is 98.5% be mixed with into protein-active thing, adds protein-active thing 0.015g in the elementary material of maize straw, hybrid reaction 3 hours at the temperature of 40 DEG C, maize straw is finally expected;
(2) added during maize straw is finally expected by cellulase 2.5g and be hydrolyzed, hydrolysis time is 48 hours, can obtain cellulose hydrolysis sugar.
Embodiment three
Use cellulase to prepare a method for cellulose hydrolysis sugar, its preparation method is as follows:
(1) pre-treatment of raw material:
1. after taking the oven dry of paddy stalk, raw material 50g and volumetric molar concentration are the aqueous sodium hydroxide solution 990g of 1%;
2. paddy stalk is soaked 2 hours in the aqueous sodium hydroxide solution of 120 DEG C;
3. soak after terminating, paddy stalk is taken out, use washed with de-ionized water corn stalk raw material until PH is 7.0, then add after the pH value of paddy stalk is adjusted to 4.0 by citric acid and obtain the elementary material of paddy stalk;
4. alpha-lactalbumin 0.9% is got by mass percentage, bovine serum albumin 1%, peptone ratio is 98.1% be mixed with into protein-active thing, adds protein-active thing 0.02g in the elementary material of paddy stalk, hybrid reaction 3 hours at the temperature of 40 DEG C, paddy stalk is finally expected;
(2) added by cellulase 2.5g during paddy stalk is finally expected and be hydrolyzed, hydrolysis time is 48 hours, can obtain cellulose hydrolysis sugar.
The content of cellulose hydrolysis sugar that following table is embodiment one, embodiment two, embodiment three, usual manner prepare cellulose hydrolysis sugar (comparative example one and comparative example two) prepares and the consumption synopsis of cellulase, wherein, the cellulosic material quality of getting in usual manner and embodiment one, embodiment two, embodiment three, embodiment four are identical, are 50g.
| Project | The content of cellulose hydrolysis sugar | The consumption of cellulase |
| Embodiment one | 36.2g/L | 2.5g |
| Embodiment two | 35.3 g/L | 2.5g |
| Embodiment three | 35.9 g/L | 2.5g |
| Comparative example one | 30.2g/L | 5g |
| Comparative example two | 30.1g/L | 5g |
Can be reached a conclusion by upper table, adopt method of the present invention to prepare cellulose hydrolysis sugar, the consumption that can either reduce cellulase, to reduce processing cost, turn improves the output of cellulose hydrolysis sugar.
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 (4)
1. use cellulase to prepare a method for cellulose hydrolysis sugar, it is characterized by, its preparation method is as follows:
The pre-treatment of cellulosic material:
Cellulosic material is taken and volumetric molar concentration is the aqueous sodium hydroxide solution of 1% by the mass ratio of 1:18 ~ 1:22;
Cellulosic material is soaked 1-3 hour in the aqueous sodium hydroxide solution of 110-130 DEG C;
After immersion terminates, cellulosic material is taken out, use washed with de-ionized water cellulosic material until PH is 6.8-7.2, then add after the pH value of cellulosic material is adjusted to 4.0-4.5 by organic acid and obtain the elementary material of Mierocrystalline cellulose;
Actives is added in the elementary material of Mierocrystalline cellulose, hybrid reaction more than 2 hours at the temperature of 38-40 DEG C, Mierocrystalline cellulose is finally expected, the quality of described actives is the 0.01%-0.05% of cellulosic material quality, and described actives is that molecular weight is at 8000 ~ 110000 daltonian egg white mixtures;
Added by cellulase during Mierocrystalline cellulose is finally expected and be hydrolyzed, hydrolysis time is 45-50 hour, can obtain cellulose hydrolysis sugar, and the quality of described cellulase is 5% of cellulosic material quality.
2. a kind of method using cellulase to prepare cellulose hydrolysis sugar as claimed in claim 1, it is characterized by, described cellulosic material comprises maize straw, paddy stalk, bagasse, Herba Eichhorniae.
3. a kind of method using cellulase to prepare cellulose hydrolysis sugar as claimed in claim 1, it is characterized by, the quality of described actives is the 0.3%-0.5% of cellulosic material quality.
4. a kind of method using cellulase to prepare cellulose hydrolysis sugar as claimed in claim 1, it is characterized by, described egg white mixture is composed of the following components by mass percentage:
Alpha-lactalbumin 0.1-1%
Bovine serum albumin 0.1-1%
Peptone 98-99.8%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105695378A (en) * | 2016-04-28 | 2016-06-22 | 韩炳鑫 | Compound enzyme preparation for producing reducing glucose by means of degraded corn stalk |
| CN106431068A (en) * | 2016-08-31 | 2017-02-22 | 武昌首义学院 | Cement retarder and preparation method thereof |
| CN106939286A (en) * | 2017-02-27 | 2017-07-11 | 华中农业大学 | A kind of method that phosphate solubilizing bacteria PA02 solid fungicides are prepared by culture matrix of plant biomass |
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| CN102181506A (en) * | 2011-02-28 | 2011-09-14 | 陕西科技大学 | Biomass pretreatment method |
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2015
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| CN101338333A (en) * | 2008-08-12 | 2009-01-07 | 南京林业大学 | Method for conversing agricultural and forest residues to be fermentable sugars |
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| CN103608461A (en) * | 2011-03-09 | 2014-02-26 | 诺维信公司 | Methods of increasing the cellulolytic enhancing activity of a polypeptide |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695378A (en) * | 2016-04-28 | 2016-06-22 | 韩炳鑫 | Compound enzyme preparation for producing reducing glucose by means of degraded corn stalk |
| CN106431068A (en) * | 2016-08-31 | 2017-02-22 | 武昌首义学院 | Cement retarder and preparation method thereof |
| CN106431068B (en) * | 2016-08-31 | 2018-11-13 | 武昌首义学院 | A kind of cement retarder and preparation method thereof |
| CN106939286A (en) * | 2017-02-27 | 2017-07-11 | 华中农业大学 | A kind of method that phosphate solubilizing bacteria PA02 solid fungicides are prepared by culture matrix of plant biomass |
| CN106939286B (en) * | 2017-02-27 | 2020-07-10 | 华中农业大学 | Method for preparing phosphate solubilizing bacterium PA02 solid microbial inoculum by taking plant biomass as culture medium |
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