CN105779528A - Lignocellulose continuous enzymatic hydrolysis and synchronous saccharification-fermentation method - Google Patents
Lignocellulose continuous enzymatic hydrolysis and synchronous saccharification-fermentation method Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 67
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- 230000001360 synchronised effect Effects 0.000 title abstract 3
- 230000004151 fermentation Effects 0.000 claims abstract description 105
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a lignocellulose continuous enzymatic hydrolysis and synchronous saccharification-fermentation method. The method comprises the following steps: (1) pre-processing lignocellulose raw materials to obtain preprocessed raw materials; (2) continuously adding the preprocessed raw materials, cellulase, and water into an enzymatic hydrolysis tank to carry out pre-enzymatic hydrolysis, and controlling the concentration of dry substances in the enzymatic hydrolysis system in a range of 18 to 36 wt%; (3) after pre-enzymatic hydrolysis, continuously feeding the feed liquid into a fermentation tank, and adding a temperature resistant brewer's yeast to carry out synchronous saccharification and fermentation; (4) after fermentation, fully mixing the mash with an inorganic flocculant in a pipeline mixer, and then delivering the mixture to a settling tank; (5) after the settlement in the settling tank, transferring the supernate to a product separating unit, and returning the concentrated mixed liquid in the lower part to the enzymatic hydrolysis tank to participate in the enzymatic hydrolysis reactions again. The method has the advantages that continuous production can be realized, the activity and utilization rate of enzymes are improved, the yield of fermentation product is increased, the enzymes can be reused, and the costs of enzymatic hydrolysis and fermentation are reduced.
Description
Technical field
The invention belongs to biomass energy source domain, a kind of method being specifically related to lignocellulose continuous enzymolysis and simultaneous saccharification and fermentation.
Background technology
Increasing along with world population and various countries' industrialization degree improves, energy resource consumption continues to rise.Oil is the Main Resources meeting energy demand, but petroleum resources are limited, and scientist predicts the year two thousand fifty crude output and will be dropped to 5,000,000,000 barrels by 25,800,000,000 barrels (35.25 hundred million tons) of 2009.Bioenergy is as a kind of reproducible transportation fuel, it is possible to effectively reduces room temperature effect, slows down environmental pollution, changes existing unequal oil supply and demand relation simultaneously, continues to supply, have clear superiority than mineral nitrogen fuel.Compared with other bioenergy such as biodiesel, the production of alcohol fuel has been provided with considerable scale (about 17,000,000 tons of the whole world in 2002), mainly to pass through fermentative Production containing sugar substance for raw material.The dehydrated alcohol (E10) mixing 10% in the oil is possible not only to alleviation Pressure on Energy, moreover it is possible to improves octane number, improves exhaust emissions quality.In the U.S. and Brazil, alcohol fuel has been widely used as traffic fuel as oil substitutes.
The Chinese government attaches great importance to energy diversification and problem of environmental pollution, takes the incentives such as financial subsidies and deductions and exemptions of taxes, carries forward vigorously technology and the industry development of the diversification petroleum replacing energy.The appearance of " Renewable Energy Law " and National Program for Medium-to Long-term Scientific and Technological Development, greatly advances the development process of the biological liquid fuel such as biodiesel and alcohol fuel.China is planned for the year two thousand twenty, and bio-fuel consumption figure accounts for about the 15% of whole traffic fuel, it is established that have the bio-fuel industry of international competitiveness, and this brings good opportunity to develop to China's fuel ethanol industrial.Current China ethanol petrol has covered 9 provinces and cities in the whole nation, existing alcohol fuel production capacity 1,520,000 tons, and actual production alcohol fuel more than 1,800,000 tons in 2010, with Semen Maydis and Semen Tritici aestivi for primary raw material.China has a large population and a few land, cultivated land resource is in short supply, and staple food supply is nervous, will threaten the grain security of country with Semen Maydis, Semen Tritici aestivi for raw material production alcohol fuel, cause the chain reactions such as agricultural product price rise, thus China strictly control with grain be raw material alcohol fuel newly-built and expand can project.
For saccharide and amylum crops, lignocellulose belongs to non-grain raw material, and aboundresources.It can derive from agricultural wastes, such as wheat straw, corn straw, corn cob, soybean residue, bagasse etc.；Trade waste, such as slurrying and the fibrous residue in paper mill, sawdust etc.；Forestry waste；Urban waste, such as waste paper, wrapping paper etc..Lignocellulosic material accounts for the 50% of world's Biomass 10,000,000,000-500 hundred million tons according to estimates, China with regard to agricultural crop straw annual output namely up to 700,000,000 tons, go out of use in a large number and do not add the fibre object (sprig, bark, leaves, bits and waste paper etc.) of utilization there are about 500,000,000 tons/annual, only also to produce the alcohol fuel of 20,000,000 tons per year with 100,000,000 tons therein.As can be seen here, the new technology developing Production of Alcohol from Lignocellulose has good prospect.
The cost of cellulase hydrolysis process cellulase, ethanol distillation process energy consumption, and the seriality of enzymolysis process is always up the industrialized restraining factors of cellulose ethanol.CN200810189465.0 discloses the production method of a kind of cellulosic ethanol, including following steps: the culture medium containing cellulose and/or hemicellulosic material is joined in fermentation reaction still by (1)；(2) in fermentation reaction still, add cellulase, and inoculate Candida lusitaniae；(3) under the combined effect of cellulase and Candida lusitaniae, carry out simultaneous saccharification and fermentation, separate and obtain cellulosic ethanol.CN200810101314.5 discloses the method that ethanol prepared by the raw material of a kind of cellulose, and the method includes the raw material of steam blasting cellulose-containing；The product of the steam explosion obtained is mixed with enzyme, enzymolysis；The product that fermentation enzyme solution obtains.In order to reduce cellulase cost, it is good method that the cellulase in enzymatic hydrolysis and fermentation liquid carries out recycling.CN200910212693.X discloses a kind of method of performing enzyme hydrolysis on wood fiber raw material, makes raw material with the wood fibre through pretreatment, adopts segmentation to carry out enzymolysis and ultrafiltration reclaimed fibre element enzyme and beta-glucosidase.But, utilize ultrafilter membrane recycled fiber element enzyme cost high, operating process is loaded down with trivial details.
Gap enzymolysis process can be used in producing, but its enzymolysis efficiency is low, can only carry out the hydrolysis of low dry concentration, hydrolysis sugar concentration is on the low side, cause final concentration of alcohol low, affect follow-up distillation energy consumption, and also have the drawbacks such as utilization rate of equipment and installations is low, floor space is big, total motor power of agitator is big.
Summary of the invention
For the deficiencies in the prior art, a kind of method that the invention provides lignocellulose continuous enzymolysis and simultaneous saccharification and fermentation.The inventive method can realize continuous prodution, improves activity and the utilization rate of enzyme, improves fermentation product yield, while ensureing that enzyme is able to reuse, reduces enzymolysis and fermentation costs.
Lignocellulose continuous enzymolysis of the present invention and simultaneous saccharification and fermentation method, including following content:
(1) lignocellulosic material is carried out pretreatment, it is thus achieved that pretreated feedstock；
(2) being added continuously in enzymatic vessel by pretreated feedstock, cellulase and water and carry out pre-enzymolysis, the dry substance concentration of controlled enzymatic hydrolysis system is 18wt%-36wt%；
(3) after pre-enzymolysis, feed liquid continuously enters in fermentation tank, adds heatproof saccharomyces cerevisiae and carries out simultaneous saccharification and fermentation；
(4) mash after fermentation is sufficiently mixed with inorganic flocculating agent in pipe-line mixer, subsequently into settling tank；
(5) supernatant after settling tank settles enters product separation unit, and bottom concentration mixed liquor loops back and again participates in enzyme digestion reaction in enzymatic vessel.
In the inventive method, first mash after step (3) described simultaneous saccharification and fermentation can carry out cell breakage process further, as high-speed stirred pearl can be adopted to grind crush method, high-pressure homogenization crush method, super wave sound crush method and the molten crush method of enzyme etc., it is preferred to use the molten crush method of enzyme；Adopting Snailase and/or lysozyme, Snailase and/or lysozyme to add in the way of continuous stream adds, the addition controlling Snailase is 1-2mg/g dry, and the addition of lysozyme is 0.2-1.0mg/g dry.The feed liquid processed through cell breakage mixes with inorganic flocculating agent in pipe-line mixer, settles subsequently into settling tank.
Lignocellulosic material in step of the present invention (1) includes the biomass material of all cellulose, such as straw, wood flour, energy-source plant (such as switchgrass) and waste paper etc., it is preferred to corn straw.Described pretreatment mode can adopt all can improve the physics of lignocellulose enzymolysis performance, chemically and thermally chemical technology, including mechanical activation comminution, radiation, microwave, acid treatment, alkali process, steam explosion pretreatment and solvent pre-treatment, or the combination pretreatment etc. of said method, it is preferred to use dilute acid steam explosion combination pretreatment.
In step (2), the time of staying of controlled enzymatic hydrolysis feed liquid is 8-96h, it is preferable that 8-24h.Size according to enzymatic vessel effective volume, controls the addition speed of pretreated feedstock and water, and the dry substance concentration (percentage ratio of soluble solids and insolubility solid masses sum and system gross mass, lower same) making enzymatic hydrolysis system is 20wt%-30wt%.Described cellulase adopts pheron or the pheron mixture of all hydrolyzable lignocellulose components, can generate cellulase online in factory, it is possible to adopt commercial goods cellulase, as Novi believes the raw enzyme of enzyme or pool.The addition controlling cellulase makes cellulase be 5-25IU/g cellulose with cellulosic ratio in pretreated feedstock.The pH controlling pre-enzymolysis is 4.5-5.5, it is preferred to 4.8-5.2；Temperature is 45-55 DEG C, it is preferred to 48-52 DEG C.
Simultaneous saccharification and fermentation in step (3) refer to pre-enzymolysis after residual cellulose at glucose fermentation producing and ethanol or other product while continuing to the process that is hydrolyzed.Described heatproof saccharomyces cerevisiae adopts the bacterial strain of available lignocellulosic material fermentation producing and ethanol or other product being currently known, the heatproof saccharomyces cerevisiae that can tolerate 36-42 DEG C is preferably used, more preferably use heatproof saccharomyces cerevisiae described in CN200910204295.3 (Saccharomycescerevisiae) FE-B, the preserving number of this bacterial strain is CGMCCNo.2735.The training method adopting this area conventional prepares zymocyte seed liquor, and the inoculum concentration of seed liquor is 1v%-5v%.In the present invention, need not regulating pH after enzymolysis, be made directly simultaneous saccharification and fermentation, control fermentation temperature and be 35-42 DEG C, the fermentation time of staying is 12-72h.The nitrogenous source added in simultaneous saccharification and fermentation system is selected from one or more in yeast extract, peptone, Semen Maydis pulp, ammonium sulfate or carbamide etc., it is preferable that carbamide, addition is the 0.02wt%-0.2wt% of system gross mass.
Inorganic flocculating agent described in step (4) is one or more in aluminum sulfate, aluminum chloride, iron sulfate, ferrous sulfate, iron chloride etc., it is preferred to iron chloride or aluminum chloride, and the addition controlling inorganic flocculating agent is 0.1-0.5mg/ dry.First fermentation liquid is mixed in proportion at pipe-line mixer with inorganic flocculating agent, settle subsequently into settling tank, the cellulase contributing to dissolving in fermentation liquid is fully adsorbed onto on solid particulate matter, cellulase is made to be circulated back in enzymatic vessel again participate in enzyme digestion reaction with the solid particulate matter of sedimentation, thus improving cellulase reuse amount.
In fermenting cellulosic ethanol mash, cellulase be concurrently present in solid-liquid biphase in, no matter product is easily separated purification by which kind of mode of follow-up employing, all can cause cellulase loss to a certain extent and inactivation.In order to realize the part reuse of enzyme, fermentation liquid generally adopts following 3 kinds of modes to circulate: 1. fermentation liquid direct part circulating and recovering；2. fermentation liquid is after solid-liquid separation, solid phase circulating and recovering；3. fermentation liquid is after solid-liquid separation, liquid phase circulation reuse.But, the solid residue granule in fermentation liquid is below tens microns, and major part is several microns only, and solid-liquid separation is extremely difficult, is generally adopted plate-and-frame filtration and adds centrifugal combination, but energy consumption is bigger；If being made directly natural subsidence, processing the cycle longer, poor effect, enzyme can not effectively reuse.Fermentation liquid is mixed by the present invention with inorganic flocculating agent, settles subsequently into settling tank, and the temperature of control settlement tank is 35-55 DEG C, and the time of staying is 0-72h.Supernatant after sedimentation flows out from settling tank top overfall, enters into product separation unit, and the concentration mixed liquor of bottom is recycled into enzymatic vessel, controls the supernatant of product separation unit and the concentration mixeding liquid volume of circulation ratio for 1:3-3:1.The concentration mixed liquor of settling tank bottom is recycled into enzymatic vessel and continues to participate in enzyme digestion reaction, and inorganic flocculating agent also loops back enzymatic vessel with concentration mixed liquor, is favorably improved enzymolysis efficiency.
Compared with prior art, the invention have the advantages that
1, in fermentation liquid, inorganic flocculating agent is added, metal ion in inorganic flocculating agent contributes to the cellulase dissolved in the liquid phase and is adsorbed onto on solid particulate matter, cellulase is made to be circulated back in enzymatic vessel again participate in enzyme digestion reaction with the solid particulate matter of sedimentation, and inorganic flocculating agent can accelerate the rate of settling of solid particulate matter, avoid running off with finely ground particle substance at continuous operation cellulase, improve the reuse amount of cellulase.
2. cellulosic enzymolysis process is an inhomogeneous reaction, and first cellulase is diffused into the cellulosic surface of substrate and is adsorbed, and is then fermentable sugars by cellulose hydrolysis.Inorganic flocculating agent is circulated back to enzymatic vessel with concentration mixed liquor, it is possible to as activator and the enzymolysis accelerator of cellulase, improves cellulase degradation efficiency, reduces fresh fiber element enzyme and makes consumption.
3, fermentation liquid is carried out cell breakage process, it is possible to reduce cellulase simultaneously and make consumption and inorganic flocculating agent make consumption.Meanwhile, yeast cells crushes the extractum of generation and can use as the nitrogenous source of alcohol fermentation along with fermentation liquid circulation, reduces fermentation costs further.
4, in fermentation liquid, add inorganic flocculating agent, substantially increase activity and the utilization rate of enzyme, improve fermentation product yield, while ensureing that enzyme is able to reuse, reduce enzymolysis and fermentation costs.The inventive method achieves continuous prodution, improves utilization rate of equipment and installations, saves the time.
Accompanying drawing explanation
Fig. 1 is the process chart of the present invention；
Wherein, 1-enzymatic vessel, 2 simultaneous saccharification and fermentation tanks, 3-cytolysis tank, 4-pipe-line mixer, 5-settling tank, 6-product separation unit.
Fig. 2 is the structural representation of settling tank of the present invention.
Detailed description of the invention
By the examples below the inventive method is described further.In the present invention, wt% is mass fraction, and v% is volume fraction.
The lignocellulosic material that the embodiment of the present invention uses is Semen Maydis dry straw, wherein cellulose 38.2wt%, and hemicellulose 22.1wt%, lignin 20.2wt%, ash 3.9wt%, being crushed to granular size with pulverizer is 1-5mm.Adopting dilute acid steam explosion to carry out pretreatment, reaction temperature is 190 DEG C, response time 5min, and solid-to-liquid ratio is 1:2, and dilute sulfuric acid concentration is 2.0wt%.Out regulating pH with NaOH afterwards from steam blasting device is 5.0, and wherein dry substance concentration is 32wt%, and in dry, content of cellulose is 40wt%, and the pretreatment dry response rate is 96wt%.
Employing heatproof saccharomyces cerevisiae (Saccharomycescerevisiae) FE-B, the preserving number of this bacterial strain is CGMCCNo.2735.Yeast starter liquid culture medium is 2wt% glucose, 2wt% peptone and 1wt% yeast extract, and 115 DEG C of sterilizing 30min are standby.The preparation of yeast starter liquid is divided into 3 grades of cultivations: the first order utilizes inoculating loop to scrape 1-2 ring bacterium mud from inclined-plane, is linked in the 25mL seed culture medium being placed in 100mL triangular flask, 37 DEG C, 100r/m, shaken cultivation 24h；The second level is cultivated and first order culture fluid is all inoculated in the 500mL seed culture medium in 1L fermentation tank, 37 DEG C, 100r/m, cultivates 24h；The third level is cultivated and second level culture fluid is all inoculated in the 25L seed culture medium in 50L fermentation tank, 37 DEG C, 100r/min, cultivates 24h.
Embodiment 1
By the flow process shown in Fig. 1, the dischargeable capacity adopting vertical enzymatic vessel is 240L, and the dischargeable capacity of simultaneous saccharification and fermentation tank is 480L, and settling tank dischargeable capacity is 240L.Corn straw good for pretreatment, cellulase (are believed Bioisystech Co., Ltd purchased from Novi, model is Ctec2, filter paper enzyme activity 135IU/g) and water be added continuously in enzymatic vessel to carry out pre-enzymolysis in proportion, wherein the addition speed of corn straw is 4.6875kg/h, the addition speed of tap water is 0.0725kg/h, the addition speed of cellulase is be equivalent to 20IU/g cellulose after 0.09kg/h(adds), in enzymatic hydrolysis system, dry substance concentration is 30wt%, in enzymatic vessel, temperature is 52 DEG C, pH5.2, stir speed (S.S.) 50r/min, the time of staying is 48h.Simultaneously enzymolysis solution is pumped into simultaneous saccharification and fermentation tank with the speed of 5kg/h from enzymatic vessel, only goes into operation the disposable access FE-B seed liquor 24L of initial start stage in system in simultaneous saccharification and fermentation tank.Nitrogenous source adopts carbamide, is configured to the mother solution of 10wt%, is continuously added to according to the speed of 0.06kg/h.In simultaneous saccharification and fermentation tank, temperature is 37 DEG C, and stir speed (S.S.) is 100r/min, and the time of staying is 96h.Mash after simultaneous saccharification and fermentation in pipe-line mixer with inorganic flocculating agent FeCl3Mixing, adds the FeCl of 5g/L with 0.09kg/h speed stream3Solution (is equivalent to 0.5mg/g dry) after addition.
Mixed fermentation liquid is pumped into sedimentation in settling tank as shown in Figure 2 with the speed of 5kg/h, and the temperature keeping settling tank is 40 DEG C, and the time of staying is 24h.Supernatant enters into product separation unit with 5kg/h speed, and concentration mixed liquor returns enzymatic vessel with 5kg/h rate loop.After circulation starts, enzymolysis solution is pumped into simultaneous saccharification and fermentation tank with the speed of 10kg/h from enzymatic vessel, and the mash in fermentation tank flows out with the speed of 10kg/h and enters in settling tank through pipe-line mixer.
The total amount taking pretreatment corn straw is 2250kg, runs 20 days continuously.Adopt liquid chromatography detection glucose and concentration of alcohol.System run all right after 144h, the concentration of glucose in sampling detection enzymolysis solution is 5.3wt% (hydrolysis sugar is had diluting effect by the solid-liquid mixed phase looped back, lower same), and glucose yield is 55.7wt%.Concentration of alcohol in fermentation tank effluent is 6.7wt%, calculates with glucose to the yield of ethanol for 90wt%, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 76.6wt%.The consumption that always makes of cellulase is 43.2kg.
Embodiment 2
By flow process shown in Fig. 1, the dischargeable capacity adopting vertical enzymatic vessel is 360L, and the dischargeable capacity of simultaneous saccharification and fermentation tank is 360L, and settling tank dischargeable capacity is 240L.By corn straw good for pretreatment, cellulase (purchased from Ze Sheng bio tech ltd, Shandong, model is 1300, filter paper enzyme activity 100IU/g) and water be added continuously in enzymatic vessel to carry out pre-enzymolysis in proportion, wherein the addition speed of corn straw is 3.125kg/h, the addition speed of tap water is 1.675kg/h, cellulase (is equivalent to 20IU/g cellulose) addition speed after addition is 0.08kg/h, in enzymatic hydrolysis system, dry substance concentration is 20wt%, in enzymatic vessel, temperature is 48 DEG C, pH4.8, stir speed (S.S.) 50r/min, the time of staying is 72h.Simultaneously enzymolysis solution is pumped into simultaneous saccharification and fermentation tank with the speed of 5kg/h from enzymatic vessel, only goes into operation the disposable access FE-B seed liquor 18L of initial start stage in system in simultaneous saccharification and fermentation tank.Nitrogenous source adopts carbamide, is configured to the mother solution of 10wt%, is continuously added to according to the speed of 0.06kg/h.In simultaneous saccharification and fermentation tank, temperature is 37 DEG C, and stir speed (S.S.) is 100r/min, and the time of staying is 72h.After simultaneous saccharification and fermentation mash in pipe-line mixer with inorganic flocculating agent AlCl3Mixing, adds the AlCl of 5g/L with 0.06kg/h speed stream3Solution (is equivalent to 0.5mg/g dry) after addition.
Mixed fermentation liquid is pumped in settling tank with the speed of 5kg/h and settles, and the temperature keeping settling tank is 40 DEG C, and the time of staying is 24h.Supernatant enters into separative element with 5kg/h speed, and concentration mixed liquor returns enzymatic vessel with 5kg/h rate loop.After circulation starts, enzymolysis solution is pumped into simultaneous saccharification and fermentation tank with the speed of 10kg/h from enzymatic vessel, and the mash in fermentation tank flows out with the speed of 10kg/h and enters in settling tank through pipe-line mixer.
The total amount taking pretreatment corn straw is 1500kg, runs 20 days continuously.System run all right after 144h, in sampling detection enzymolysis solution, concentration of glucose is 3.6wt%, and glucose yield is 64.8wt%.In fermentation tank effluent, concentration of alcohol is 4.0wt%, calculates with glucose to the yield of ethanol for 90wt%, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 78.4wt%.Cellulase makes consumption be 38.4kg.
Embodiment 3
Handling process is identical with embodiment 1 with process conditions, is different in that: the addition speed of enzymatic vessel cellulase is be equivalent to 15IU/g cellulose after 0.0675kg/h(adds), it is 0.125kg/h that tap water adds speed.Yeast is carried out cell breakage initially with Snailase by fermentation liquid, the cytolysis tank time of staying is 8h, and Snailase is configured to the aqueous solution of 1g/L, and addition speed is 0.03kg/h (being equivalent to 2mg/g dry after addition), temperature is 37 DEG C, and pH is 6.0.Subsequently in pipe-line mixer with inorganic flocculating agent FeCl3Mixing, adds the FeCl of 5g/L with 0.06kg/h speed stream3Solution, makes FeCl3Mix homogeneously with the fermentation liquid after cell breakage (after addition, being equivalent to 0.2mg/g dry).Mixed mash is pumped into sedimentation in settling tank as shown in Figure 2 with the speed of 5kg/h, and the temperature keeping settling tank is 40 DEG C, and the time of staying is 24h.Enter in enzymatic vessel with 5kg/h rate loop through the solidliquid mixture of concentration bottom settling tank.After circulation starts, enzymolysis solution is pumped into simultaneous saccharification and fermentation tank with the speed of 10kg/h from enzymatic vessel, fermentation liquid is pumped into settling tank with the speed of 10kg/h from simultaneous saccharification and fermentation tank, the mash that the dry concentration on settling tank top reduces exits into product separation unit with the speed of 5kg/h, stopping stream in fermentation tank simultaneously and add urea liquid, in enzymatic vessel, tap water rate adaptation is 0.185kg/h.
The total amount taking pretreatment corn straw is 2250kg, runs 20 days continuously.System run all right after 144h, the concentration of glucose in sampling detection enzymolysis solution is 6.3wt%, and glucose yield is 66.2wt%.Concentration of alcohol in fermentation tank effluent is 7.4wt%, calculates with the yield of glucose to ethanol 90%, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 84.6wt%.The consumption that makes of cellulase is 32.4kg.
Embodiment 4
Handling process is identical with embodiment 2 with process conditions, is different in that: the addition speed of enzymatic vessel cellulase is be equivalent to 15IU/g cellulose after 0.06kg/h(adds), it is 1.715kg/h that tap water adds speed.Yeast is crushed by simultaneous saccharification and fermentation mash initially with Snailase, in cytolysis tank, the time of staying is 8h, and Snailase is configured to the aqueous solution of 1g/L, and addition speed is 0.02kg/h (being equivalent to 2mg/g dry after addition), temperature is 37 DEG C, and pH is 6.0.Subsequently in pipe-line mixer with inorganic flocculating agent AlCl3Mixing, adds the AlCl of 5g/L with 0.04kg/h speed stream3Solution, makes AlCl3Mix homogeneously with fermentation liquid (after addition, being equivalent to 0.2mg/g dry).Mixed mash is pumped into sedimentation in settling tank as shown in Figure 2 with the speed of 5kg/h, and the temperature keeping settling tank is 40 DEG C, and the time of staying is 24h.Enter in enzymatic vessel with 5kg/h rate loop through the solidliquid mixture of concentration bottom settling tank.After circulation starts, enzymolysis solution is pumped into simultaneous saccharification and fermentation tank with the speed of 10kg/h from enzymatic vessel, fermentation liquid is pumped into settling tank with the speed of 10kg/h from simultaneous saccharification and fermentation tank, the mash that the dry concentration on settling tank top reduces exits into product separation unit with the speed of 5kg/h, stopping stream in fermentation tank simultaneously and add urea liquid, in enzymatic vessel, tap water rate adaptation is 1.775kg/h.
The total amount of the corn straw processed is 1500kg, runs 20 days continuously.System run all right after 144h, the concentration of glucose in sampling detection enzymolysis solution is 3.7%, and pre-enzymolysis stage glucose yield is 66.6%.Concentration of alcohol in fermentation tank effluent is 4.4%, is 90% calculating with glucose to the yield of ethanol, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 86.3%.The consumption that makes of cellulase is 28.8kg.
Embodiment 5
Handling process is identical with embodiment 3 with process conditions, is different in that: adopting high-pressure homogenization disintegrating machine that fermentation liquid carries out cell breakage, adopt continuous operation mode, cracking pressure is 50MPa, and controlling outlet temperature is 20 DEG C.After system run all right, the concentration of glucose in sampling detection enzymolysis solution is 5.7wt%, and glucose yield is 59.9wt%.Concentration of alcohol in fermentation tank effluent is 7.0wt%, is 90% calculating with glucose to the yield of ethanol, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 80.1wt%.The consumption that makes of cellulase is 32.4kg.
Comparative example 1
Handling process is identical with embodiment 1 with process conditions, is different in that: the fermentation liquid after fermentation ends does not enter settling tank, directly enters product separation unit according to the proportional parts of 1:1, and part loops back in enzymatic vessel.After system run all right, the concentration of glucose in sampling detection enzymolysis solution is 4.9wt%, and glucose yield is 51.5wt%.Concentration of alcohol in fermentation tank effluent is 6.1wt%, is 90% calculating with glucose to the yield of ethanol, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 73.2wt%.The consumption that makes of cellulase is 43.2kg.
Comparative example 2
Handling process is identical with embodiment 1 with process conditions, is different in that: the fermentation liquid after fermentation ends enters solid-liquid separation tank and carries out solid-liquid separation, and isolated liquid phase enters product separation unit, and isolated solid phase loops back in enzymatic vessel.After system run all right, the concentration of glucose in sampling detection enzymolysis solution is 4.8wt%, and glucose yield is 50.4wt%.Concentration of alcohol in fermentation tank effluent is 5.9wt%, is 90% calculating with glucose to the yield of ethanol, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 72.1%.The consumption that makes of cellulase is 43.2kg.
Comparative example 3
Handling process is identical with embodiment 3 with process conditions, is different in that: be added without inorganic flocculating agent, only carries out cell breakage.System run all right after 72h, the concentration of glucose in sampling detection enzymolysis solution is 5.8wt%, and glucose yield is 60.9wt%.Concentration of alcohol in fermentation tank effluent is 6.7wt%, is 90% calculating with glucose to the yield of ethanol, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 76.6wt%.The consumption that makes of cellulase is 32.4kg.
Comparative example 4
Handling process is identical with embodiment 4 with process conditions, is different in that: be added without inorganic flocculating agent, only carries out cell breakage.System run all right after 72h, the concentration of glucose in sampling detection enzymolysis solution is 3.5%, and glucose yield is 63.0%.Concentration of alcohol in fermentation tank effluent is 4.1%, is 90% calculating with glucose to the yield of ethanol, and the enzymolysis of whole system and the glucose yield of simultaneous saccharification and fermentation are 80.4%.The consumption that makes of cellulase is 28.8kg.
Claims (11)
1. the method for a lignocellulose continuous enzymolysis and simultaneous saccharification and fermentation, it is characterised in that include following content:
(1) lignocellulosic material is carried out pretreatment, it is thus achieved that pretreated feedstock；
(2) being added continuously in enzymatic vessel by pretreated feedstock, cellulase and water and carry out pre-enzymolysis, controlled enzymatic hydrolysis system dry substance concentration is 18wt%-36wt%；
(3) after pre-enzymolysis, feed liquid continuously enters in fermentation tank, adds heatproof saccharomyces cerevisiae and carries out simultaneous saccharification and fermentation；
(4) mash after fermentation is sufficiently mixed with inorganic flocculating agent in pipe-line mixer, subsequently into settling tank；
(5) supernatant after settling tank settles enters product separation unit, and bottom concentration mixed liquor loops back and again participates in enzyme digestion reaction in enzymatic vessel.
2. in accordance with the method for claim 1, it is characterised in that: the mash after step (3) described simultaneous saccharification and fermentation carries out cell breakage process, adopts high-speed stirred pearl to grind crush method, high-pressure homogenization crush method, super wave sound crush method or the molten crush method of enzyme.
3. in accordance with the method for claim 2, it is characterised in that: cell breakage adopts the molten crush method of enzyme, uses Snailase and/or lysozyme, adds in the way of continuous stream adds.
4. in accordance with the method for claim 3, it is characterised in that: the addition controlling Snailase is 1-2mg/g dry, and the addition of lysozyme is 0.2-1.0mg/g dry.
5. in accordance with the method for claim 1, it is characterised in that: in step (1), described pretreatment adopts dilute acid steam explosion combination pretreatment.
6. in accordance with the method for claim 1, it is characterised in that: in step (2), the time of staying of controlled enzymatic hydrolysis feed liquid is 8-96h；The addition controlling cellulase makes cellulase be 5-25IU/g cellulose with cellulosic ratio in pretreated feedstock；The pH of pre-enzymolysis is 4.5-5.5, and temperature is 45-55 DEG C.
7. in accordance with the method for claim 1, it is characterised in that: in step (3), heatproof saccharomyces cerevisiae use heatproof saccharomyces cerevisiae described in CN200910204295.3 (Saccharomycescerevisiae) FE-B, the preserving number of this bacterial strain is CGMCCNo.2735.
8. the method described in claim 1 or 7, it is characterised in that: the inoculum concentration of heatproof saccharomyces cerevisiae seed liquor is 1v%-5v%；Need not regulating pH after enzymolysis, be made directly simultaneous saccharification and fermentation, control fermentation temperature and be 35-42 DEG C, the fermentation time of staying is 12-72h.
9. in accordance with the method for claim 1, it is characterized in that: the inorganic flocculating agent described in step (4) is one or more in aluminum sulfate, aluminum chloride, iron sulfate, ferrous sulfate, iron chloride, the addition controlling inorganic flocculating agent is 0.1-0.5mg/g dry.
10. in accordance with the method for claim 1, it is characterised in that: the temperature of step (5) control settlement tank is 35-55 DEG C, and the time of staying is 0-72h.
11. the method described in claim 1 or 10, it is characterised in that: step (5) controls the concentration mixeding liquid volume of the supernatant of product separation unit and circulation ratio for 1:3-3:1.
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| CN109628542A (en) * | 2018-12-29 | 2019-04-16 | 中南林业科技大学 | A method of extracting bran polysaccharide from wheat bran |
| CN109971347A (en) * | 2019-03-06 | 2019-07-05 | 常州市三伊化工有限公司 | A kind of preparation method of high viscosity high intensity raw lacquer |
| CN111500291A (en) * | 2020-04-22 | 2020-08-07 | 张丽莉 | Enzyme catalysis conditioning agent for soil remediation and preparation method and application thereof |
| CN117625387A (en) * | 2023-12-04 | 2024-03-01 | 哈尔滨工业大学 | Three-phase conveying type straw particle enzymolysis multistage fermentation hydrogen production device and fermentation hydrogen production method thereof |
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| CN102260139A (en) * | 2010-05-24 | 2011-11-30 | 安徽丰原生物化学股份有限公司 | Separation method of alcoholic fermented liquor |
| CN103103217A (en) * | 2011-11-10 | 2013-05-15 | 中国石油化工股份有限公司 | Production method for fuel ethanol |
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| CN102260139A (en) * | 2010-05-24 | 2011-11-30 | 安徽丰原生物化学股份有限公司 | Separation method of alcoholic fermented liquor |
| CN103103217A (en) * | 2011-11-10 | 2013-05-15 | 中国石油化工股份有限公司 | Production method for fuel ethanol |
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| CN109628542A (en) * | 2018-12-29 | 2019-04-16 | 中南林业科技大学 | A method of extracting bran polysaccharide from wheat bran |
| CN109628542B (en) * | 2018-12-29 | 2020-11-24 | 中南林业科技大学 | Method for extracting bran polysaccharide from bran |
| CN109971347A (en) * | 2019-03-06 | 2019-07-05 | 常州市三伊化工有限公司 | A kind of preparation method of high viscosity high intensity raw lacquer |
| CN111500291A (en) * | 2020-04-22 | 2020-08-07 | 张丽莉 | Enzyme catalysis conditioning agent for soil remediation and preparation method and application thereof |
| CN111500291B (en) * | 2020-04-22 | 2024-01-19 | 张丽莉 | Enzyme-catalyzed conditioner for restoring soil and preparation method and application thereof |
| CN117625387A (en) * | 2023-12-04 | 2024-03-01 | 哈尔滨工业大学 | Three-phase conveying type straw particle enzymolysis multistage fermentation hydrogen production device and fermentation hydrogen production method thereof |
| CN117625387B (en) * | 2023-12-04 | 2024-10-01 | 哈尔滨工业大学 | Three-phase conveying type straw particle enzymolysis multistage fermentation hydrogen production device and fermentation hydrogen production method thereof |
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