CN101037703B - Hydrolysis and fermentation method for reinforcing straw cellulose - Google Patents
Hydrolysis and fermentation method for reinforcing straw cellulose Download PDFInfo
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- CN101037703B CN101037703B CN2006100651027A CN200610065102A CN101037703B CN 101037703 B CN101037703 B CN 101037703B CN 2006100651027 A CN2006100651027 A CN 2006100651027A CN 200610065102 A CN200610065102 A CN 200610065102A CN 101037703 B CN101037703 B CN 101037703B
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Abstract
The invention relates to a enzymatic hydrolysis and fermentation method of reinforcing straw cellulose, using crops steam-exploded straw as material, adding cellulase and rust ball in solid fermentation reactor, in swaying dynamic condition which pH=4-6 at 30-50 DEG C, enzymolysis steam-exploded straw at high speed to generate reductant sugar; the added cellulase is commercial Aspergillus niger; relationship between steam-exploded straw amount and cellulase amount: adding 11 IU cellulase in each gram of crops steam-exploded straw. Also adding saccharomycete to generate ethyl; relationship between steam-exploded straw amount and saccharomycete amount is: adding 0.01 g yeast in each gram of reinforcing straw cellulose. Shaking the reactor to revolve stainless steel ball to reinforce mass transfer, raise saccharification, ferment reaction rate. The invention is applied mainly in cellulose solid fermentation, can raise greatly steam-exploded straw saccharification, ferment reaction rate and solve energy inversion problem of biomass.
Description
Technical field
The invention belongs to the steam puffed stalk manufacture field, the cellulosic hydrolysis and fermentation method of particularly a kind of reinforcement steam puffed stalk.
Background technology
Along with the rapid growth of population, crisis of resource, be short of food, problem such as environmental pollution more and more seriously perplexing human society, caused that people pay close attention to widely.8,666 hundred million tons of plant organic matters of annual approximately formation on the earth, be that distributed in nature is the widest, content is maximum, cheap, and the renewable resources that is underutilized, except that being used for industries such as papermaking, building, weaving on a small quantity or with producing fodder, the fuel wood, major part is not used effectively, and rot in vain or burn, some also causes environmental pollution (for example burning).By biotechnology these fiber wastes are converted into the energy, Chemicals etc., will be one of human approach that solves following grain, the energy and industrial chemicals problem, of far-reaching significance..
Our used fossil oil (coal, oil and natural gas) all is the remains of ancient times biomass now.How many accumulation in 100000000 years, the modern who has been grasped scientific and technological ability used totally in two to 300 years.In the reproducible on earth green resource, cellulosic source is maximum, but people are very limited to the development and utilization of wood-cellulose, and the degradation mechanism of failing to get clear natural cellulose is a major reason.For a long time, mainly concentrate on the mould cellulase hydrolysis effect aspect that waits filamentous fungus of wood about the cellulose degradation Study on Mechanism, now got the mechanism of they hydrolysis crystalline celluloses substantially clear, but all be that material carries out, still fail to illustrate the reason that natural wooden fiber's element is difficult to be degraded with the crystalline cellulose.
In natural wooden fiber's material, owing to tightly link to each other between hemicellulose and lignin and Mierocrystalline cellulose, particularly the lignin bag is by fiber, and cellulase can't contact with fiber, and the enzymic hydrolysis effect can not be carried out, and this is the biggest obstacle during lignocellulosic material utilizes.For improving enzymolysis efficiency, the general method that at first stalk is adopted and handle improves the accessibility and the reactive behavior of fiber.
At present, the pretreatment process of stalk generally speaking can be divided three classes: physics method, chemical method and biological process.The physics method comprises grinding, pulverizing, high temperature, steaming and decocting under high pressure, explosion, microwave, radiotreatment etc.; Chemical method comprises acid, alkali, organic solvent method; Biological process mainly is microbial method and enzyme process.This several method cuts both ways.Consider from environment, energy consumption, hydrolysis result equal angles, the quick-fried technology of vapour is to be everybody confessed best pretreatment process up to now, steam explosion is that a kind of elder generation is with raw material vapor phase pulping for some time under high temperature, high-pressure medium, make hemicellulose be degraded into soluble sugar, xylogen softens and the part degraded simultaneously, reduces fiber bond intensity, unexpected again release of pressure, make material be dissociated into single fibrocyte, increase the accessibility of raw material greatly from middle lamella.Discover that the stalk after vapour is quick-fried has stronger infiltration, diffusion and swollen performance, porosity also improves greatly.
But the stalk enzymolysis speed through the quick-fried processing of vapour is still lower, can not satisfy industrial requirement, apart from commercial application certain distance is arranged still.So,, will provide the feasibility reference for the enzymolysis of steam puffed stalk, the industrialization of fermentation if can under the prerequisite that does not increase new environmental disruption factor, increase substantially the enzymolysis speed of steam puffed stalk.
Amount of literature data studies show that the low cellulosic speed of enzyme hydrolysis of temperature is slow, and the temperature height causes the inactivation of enzyme easily, and cellulosic enzymic hydrolysis temperature is more suitable in 40-55 ℃ scope; Cellulosic enzymic hydrolysis is very sensitive to the variation of pH value, and comparatively the pH value of Shi Heing is 4.8-5.2, and when in this pH and temperature range Mierocrystalline cellulose being carried out enzymolysis, fermentation reaction, enzyme is lived higher, can obtain higher enzymolysis speed.
Enzymatic hydrolysis and fermentation speed for resistant cellulose, someone has studied ultrasonic wave influences cellulosic enzymic hydrolysis, discover: ultrasonic wave is in the plain enzymic hydrolysis of the ultrasonic energy accelerating fibers of certain frequency and strength range, and it is relevant with frequency and intensity, hyperacoustic influence can be uncovered the effect of the enzymolysis that quickens at short notice, and the fugitive more fruit of hydrolysis time is obvious more.Simultaneously, discover that ultrasonic wave plays deactivation to cellulase.
In the research of cellulase hydrolysis and fermentation at home and abroad,, in enzymolysis, fermentation, adopt relatively difficulty of dynamical fashion because cellulosic materials such as steam puffed stalk are different from general granular solid state fermentation raw material.So up to now, domestic and international research all is to adopt the mode that leaves standstill constant temperature enzymolysis, fermentation to carry out basically.
Summary of the invention
It is lower to the present invention is directed to present steam puffed stalk enzymolysis speed, proposition a kind of efficiently, green steam puffed stalk enzymatic hydrolysis and fermentation production technique.Use stalk for making raw material, the pretreatment mode combination with a series of clean and effectives makes stalk have stronger infiltration, diffusion and swollen performance, and porosity also improves greatly; Then, carry out the method for enzymatic hydrolysis and fermentation by dynamic enzymolysis process novel, environmental protection.
Technical scheme of the present invention is as follows:
The hydrolysis and fermentation method of reinforcing straw cellulose provided by the invention,
At first agricultural crop straw is soaked by a certain percentage, in the quick-fried device of the vapour of packing into, under the pressure of 1.0～2.0MPa, keep 1～15min after, fast release of pressure sprays stalk from steam-explosion jar, obtains steam puffed stalk;
With the crops steam-exploded straw is raw material, adds cellulase and rust system globe in solid-state fermentation reactor, and in pH=4～6, temperature is 30-50 ℃, under the dynamic condition that rocks, makes steam puffed stalk high speed enzymolysis produce reducing sugar; The cellulase that is added is a commercial Aspergillus niger; Described crops steam-exploded straw with the quantity relative ratio relationship of the cellulase that is added is: every gram crops steam-exploded straw adds the 11IU cellulase.
Described crops steam-exploded straw is or/and straw through the corn stalk of the quick-fried processing of vapour, wheat straw, hybrid giant napier.The described dynamic condition that rocks is meant that solid-state fermentation reactor shakes with 50r/min～500r/min.Described rust system globe is the stainless steel steel ball.
The synchronous glycosylation fermentation process of reinforcing straw cellulose provided by the invention adds yeast when adding cellulase, and then generates ethanol; Described crops steam-exploded straw and the saccharomycetic quantity relative ratio relationship that is added are: every gram reinforcing straw cellulose adds the 0.01g yeast.
The present invention with the agriculture and forestry organic waste material is raw material, and is with low cost, broken through at present the only idea of enzymolysis, fermentation constraint under static conditions of steam puffed stalk in the world; Adopting efficient enzymolysis pretreatment mode---on the basis of the quick-fried technology of vapour, stalk after utilizing vapour quick-fried has stronger infiltration, diffusion and swollen performance, characteristic that porosity is big, use Stainless Steel Ball as special mass transfer instrument first, the method that Stainless Steel Ball is rocked at a high speed increases the mass transfer of enzymolysis, fermentation, thereby significantly improves enzymolysis speed mode.Present method adopts the cleaning steam explosion and combines with dynamic enzymolysis, the efficient height, and omnidistance pollution-free, enzyme activity is loss not, has great industrial applications value.
Embodiment
Below by embodiment the present invention is further specified:
The application of embodiment 1 the present invention in the steam exploded wheat straw enzymolysis process
The wheat straw of regulating water content to 15～35% is packed in the quick-fried device of vapour, and after logical saturation steam reached 1.5MPa pressure, dimension was pressed 10min, relief pressure suddenly, steam exploded wheat straw;
The steam exploded wheat straw that obtains is taken out,, carries out enzymolysis after the taking-up 85 ℃ of following continuous countercurrent washing desugars:
In Plastic Bottle, add steam exploded wheat straw 10g (dry weight), liquid cellulase 1ml (FPA=110IU/ml), sodium-acetate-acetate buffer solution 100ml, under the condition of pH=4.8, enzyme digestion reaction under different mass transfer modes:
1. static enzymolysis in the water-bath;
2. vibration enzymolysis (adding one piece of Stainless Steel Ball simultaneously) in the shaking table, the rotating speed of shaking table is respectively 50r/min, 130r/min, 210r/min, 500r/min
After the enzymolysis 1hr time, suction filtration records the sugared concentration and the output of sugar of enzymolysis solution respectively and compares result such as table 1;
Table 1 is under different enzymolysis modes, the output of sugar (pipe) of the steam exploded wheat straw of enzymolysis after 1 hour
From table 1, learn: in 50r/min～500r/min speed range, adopt the output of sugar of steam exploded wheat straw enzymolysis all to be higher than and leave standstill enzymolysis in the steel ball mode of oscillation.As seen, under 50 ℃, the output of sugar that records behind the 1hr under the steel ball vibration enzymolysis mode (210r/min) is 0.9g in the table, and the mode output of sugar of leaving standstill is 0.3g, shows that interior the former output of sugar of 1hr is 3 times of the latter;
The application of embodiment 2 the present invention in steam exploded wheat straw synchronous glycosylation fermenting process
The wheat straw of regulating water content to 15～35% is packed in the quick-fried device of vapour, and after logical saturation steam reached 1.5MPa pressure, dimension was pressed 10min, and relief pressure takes out 85 ℃ of continuous countercurrents and washes desugars suddenly; Carry out diastatic fermentation after the taking-up.
In Plastic Bottle, add steam exploded wheat straw 10g (dry weight) respectively, Angel Yeast 0.1g, liquid cellulase 1ml (FPA=110IU/ml), sodium-acetate-acetate buffer solution 100ml, at 40 ℃, under the condition of pH=4.8, enzyme digestion reaction under different mass transfer modes: 1. static enzymolysis in the water-bath; 2. vibration (adding one piece of Stainless Steel Ball simultaneously) in the shaking table is vibrated under 50r/min, 130r/min, 210r/min, 210r/min respectively.Ferment after 24 hours, the alcohol output that records enzymolysis solution respectively compares.Its result such as table 2;
Table 2 is different saccharification fermentation modes, the alcohol output of the steam exploded wheat straw after 24 hours (v/v) influence
From table 2 as seen: in the speed range of 50r/min～500r/min, adopt under the steel ball mode of oscillation alcohol output that steam exploded wheat straw is carried out diastatic fermentation all to be higher than the mode of leaving standstill.As seen, under 50 ℃, record the alcohol output 0.90% (v/v) under the steel ball mode of oscillation (210r/min) behind the 24hr in the table, and the alcohol output that leaves standstill mode is 1.8% (v/v), the former alcohol amount is 2.0 times of the latter.
The application of embodiment 3 the present invention in the quick-fried straw enzymolysis process of vapour
The straw of regulating water content to 15～35% is packed in the quick-fried device of vapour, and after logical saturation steam reached 1.5MPa pressure, dimension was pressed 10min, relief pressure suddenly, the quick-fried straw of vapour;
The quick-fried straw of the vapour that obtains is taken out,, carries out enzymolysis after the taking-up 85 ℃ of following continuous countercurrent washing desugars:
In Plastic Bottle, add the quick-fried straw 10g of vapour (dry weight), liquid cellulase 1ml (FPA=110IU/ml), sodium-acetate-acetate buffer solution 100ml, under the condition of pH=4.8, enzyme digestion reaction under different mass transfer modes:
3. static enzymolysis in the water-bath;
4. vibration enzymolysis (adding one piece of Stainless Steel Ball simultaneously) in the shaking table, the rotating speed of shaking table is respectively 50r/min, 130r/min, 210r/min, 500r/min
After the enzymolysis 1hr time, suction filtration records the sugared concentration and the output of sugar of enzymolysis solution respectively and compares result such as table 3;
Table 3 is under different enzymolysis modes, the output of sugar (g) of the vapour quick-fried straw of enzymolysis after 1 hour
From table 3, learn: in 50r/min～500r/min speed range, adopt the output of sugar of the quick-fried straw enzymolysis of vapour all to be higher than and leave standstill enzymolysis in the steel ball mode of oscillation.As seen, under 50 ℃, the output of sugar that records behind the 1hr under the steel ball vibration enzymolysis mode (210r/min) is 1.03g in the table, and the mode output of sugar of leaving standstill is 0.34g, shows that interior the former output of sugar of 1hr is 3.03 times of the latter.
Claims (5)
1. the hydrolysis and fermentation method of a reinforcing straw cellulose, it is characterized in that, with the crops steam-exploded straw is raw material, in solid-state fermentation reactor, add cellulase and Stainless Steel Ball, in pH=4～6, temperature is 30-50 ℃, under the dynamic condition that rocks, makes steam exploded wheat straw high speed enzymolysis produce reducing sugar; Described crops steam-exploded straw with the quantity relative ratio relationship of the cellulase that is added is: every gram crops steam-exploded straw adds the 11IU cellulase.
2. by the hydrolysis and fermentation method of the described reinforcing straw cellulose of claim 1, it is characterized in that described crops steam-exploded straw is or/and straw through the corn stalk of the quick-fried processing of vapour, wheat straw, hybrid giant napier.
3. by the hydrolysis and fermentation method of the described reinforcing straw cellulose of claim 1, it is characterized in that the described dynamic condition that rocks is meant that solid-state fermentation reactor shakes with 50r/min～500r/min.
4. by the hydrolysis and fermentation method of the described reinforcing straw cellulose of claim 1, it is characterized in that described rust system globe is the stainless steel steel ball.
5. want the synchronous glycosylation fermentation process of 1,2,3 or 4 described reinforcing straw celluloses by right, it is characterized in that, when adding cellulase, add yeast, and then generate ethanol; Described crops steam-exploded straw and the saccharomycetic quantity relative ratio relationship that is added are: every gram reinforcing straw cellulose adds the 0.01g yeast.
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101768545B (en) * | 2010-02-24 | 2013-11-06 | 中国科学院过程工程研究所 | Cycling elution biomimetic peristaltic enzymolysis reactor and method thereof |
| AU2010361837B2 (en) * | 2010-09-29 | 2015-01-29 | Versalis S.P.A. | Pre-treated biomass having enhanced enzyme accessibility |
| CN103289903B (en) * | 2013-05-07 | 2015-02-04 | 山西省农业科学院农业环境与资源研究所 | Aspergillus niger H active decomposition accelerator with bacteriostasis function and preparation method thereof |
| CN103436586B (en) * | 2013-07-18 | 2015-08-26 | 济南开发区星火科学技术研究院 | One utilizes fermentable biomass to produce alcohol technique |
| CN103710394B (en) * | 2013-12-10 | 2016-05-11 | 南昌大学 | A kind of method of utilizing oil-extracted algae slag to produce bio-ethanol |
| CN105441490A (en) * | 2015-12-05 | 2016-03-30 | 贵州省欣紫鸿药用辅料有限公司 | Production method for preparing medical ethyl alcohol through corn straw |
| CN105695533A (en) * | 2016-03-09 | 2016-06-22 | 孟繁志 | Additive for preparation of cellulose and preparation method of cellulose |
| CN109024031A (en) * | 2018-10-12 | 2018-12-18 | 山东金胜粮油集团有限公司 | The method that steam blasting combination ultrasonic treatment prepares peanut shell nano-cellulose |
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| CN1451755A (en) * | 2002-04-12 | 2003-10-29 | 中国农业大学 | Method for producing alcohol by solid fermentation of stalks |
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| CN1451755A (en) * | 2002-04-12 | 2003-10-29 | 中国农业大学 | Method for producing alcohol by solid fermentation of stalks |
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| 孙智谋.混合酶及汽爆法提高秸杆发酵酒精的产量.酿酒科技 5.2004,(5),75-78. |
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