CN105132470B - A kind of method that enzyme recycles in lignocellulosic enzymolysis process - Google Patents
A kind of method that enzyme recycles in lignocellulosic enzymolysis process Download PDFInfo
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- CN105132470B CN105132470B CN201510636139.XA CN201510636139A CN105132470B CN 105132470 B CN105132470 B CN 105132470B CN 201510636139 A CN201510636139 A CN 201510636139A CN 105132470 B CN105132470 B CN 105132470B
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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
Abstract
The invention belongs to the technical fields of cellulose ethanol production.A kind of method that enzyme recycles in lignocellulosic enzymolysis process, specific step is as follows: starting enzymatic vessel, and according to the concentration of substrate of setting, material, moisture batch continuous uniform are added in enzymatic vessel, continuous uniform adds lignocellulosic hydrolase in batches simultaneously, is digested；After enzymatic hydrolysis, enzymolysis liquid is cooled to 30 ~ 32 DEG C, saccharomyces cerevisiae is inoculated with, ferments；Gained fermentation liquid imports in vacuum distillation tower and carries out vacuum distillation；Vinasse obtained by vacuum distillation imports and carries out enzyme reuse in enzymatic vessel, and cycle-index is 1 ~ 2 time.The present invention passes through vacuum distillation, cellulase activity remaining in fermentation liquid is remained, vinasse is added in new enzymatic hydrolysis system by reuse, 20 ~ 25% lignocellulosic hydrolysis enzyme dosage can be reduced, the utilization rate of cellulase is greatly improved, to reduce production cost.
Description
Technical field
The invention belongs to the technical fields of cellulose ethanol production, and in particular to enzyme follows in a kind of lignocellulosic enzymolysis process
The method that ring utilizes.
Background technique
Lignocellulose raw material is renewable resource most abundant, most cheap on the earth.The whole world passes through photosynthesis every year
The wood fiber biomass of generation is up to 100,000,000,000 tons.China is large agricultural country, has biomass resource abundant, only crops
Stalk one just has nearly 8 × 10 every year8Ton (being roughly equal to 400,000,000 tons of mark coals), the 10 ~ 15% of Zhan Quanguo primary energy consumption total amount, add
The forestry fiber waste and industrial fiber waste of upper enormous amount, annual available lignocellulose raw material total amount is up to 2,000,000,000 tons
More than.Cellulose accounts for about the 35 ~ 45% of dry weight in lignocellulose raw material, and hemicellulose accounts for about 20 ~ 40%, will using appropriate technology
They are hydrolyzed into fermentable sugar, and further fermentative production of ethanol makes full use of these biomass resources to can be relieved china natural resources
The pressure of shortage mitigates environmental pollution, pushes Agro-ecology benign cycle, and to construction, energy-saving low-carbon society is of great significance.
The production process of usual cellulose ethanol includes pretreatment, enzyme hydrolysis, fermentation and the distilation of raw material, wherein digesting
It is the major technique and economical one of entire cellulose ethanol production.Currently, the most important problem of cellulose hydrolysis and saccharification is enzymatic hydrolysis
Low efficiency, the enzymatic hydrolysis period is long, and enzyme dosage is big, and the not high aspect of conversion ratio, these problems seriously constrain lignocellulose raw material
Bioconversion and utilization.The costly of cellulase preparation is one of the principal element for influencing bio-ethanol price, reduces fiber
The technology of plain enzyme usage amount or expense becomes bio-ethanol and produces emphasis.
Cellulase is adsorbed on above substrate at hydrolysis of lignocellulose initial stage, when after reaction, due to substrate
The variation of concentration, enzyme are gradually detached from substrate, are become the state of resolvase, are scattered in enzymolysis liquid, and another part is then
It is adsorbed in the enzymatic hydrolysis residue of high lignin content.Resolvase in enzymolysis liquid can give full play to its valence by recycling and reusing
Value.
It realizes efficiently enzymatic hydrolysis, reduces enzymatic hydrolysis cost, can set about from following three: 1) pretreatment expected；2) it drops as far as possible
The industrial production cost of low in hydrolysis enzyme, enzyme cost are reduced to a certain range, and the entire cost of investment of cellulose ethanol can just have with petroleum
Competitive superiority；3) effective fiber element enzyme recycles and reuses, even if the enzyme during enzyme hydrolysis adequately participates in reacting, increases
The enzyme touch opportunity with cellulose, improves enzymolysis efficiency, realizes that enzyme recycles and reuses.
Patent CN101974570A discloses a kind of side of recycling cellulose complete components in fuel ethanol production
Method refers to and utilizes coupling fresh substrate heavily adsorb and immobilized cellobiase work after lignocellulosic simultaneous saccharification and fermentation workshop section
The method of skill recycling and reusing cellulase full constituent, which is related to immobilization process, due to immobilised enzymes and cellulose
There is very big mass transfer obstacle between solid particle, therefore is currently limited to cellobiase in lignocellulosic enzymatic hydrolysis system
Immobilization recycling.
Summary of the invention
The technical problem to be solved by the present invention is to underuse for traditional enzymolysis process free cellulose enzyme, mention
For the method that enzyme in a kind of lignocellulosic enzymolysis process recycles, the utilization rate of free cellulose enzyme is improved, reduces production
Cost.
In order to solve the above technical problems, the technical scheme adopted by the invention is that: in a kind of lignocellulosic enzymolysis process
The method that enzyme recycles is connected using enzymatic vessel, fermentor and destilling tower and is carried out, first digested by the feed ratio of setting,
After enzymatic hydrolysis, cooling inoculation is fermented, then fermentation liquid is imported vacuum distillation tower and carries out vacuum distillation, removes ethyl alcohol, living
Property yeast inactivation, leave free cellulase, finally will vinasse import enzymatic hydrolysis system in carry out reuse, specific steps are such as
Under:
Step 1): material, moisture batch continuous uniform are added to enzymatic hydrolysis according to the concentration of substrate of setting by starting enzymatic vessel
In tank, while continuous uniform adds lignocellulosic hydrolase in batches, is digested；
Step 2: after enzymatic hydrolysis, enzymolysis liquid is cooled to 30 ~ 32 DEG C, saccharomyces cerevisiae is inoculated with, ferments；
Step 3): step 2 fermentation gained fermentation liquid is imported in vacuum distillation tower and carries out vacuum distillation；
Step 4): vinasse obtained by step 3) vacuum distillation is imported and carries out enzyme reuse, cycle-index 1 in enzymatic vessel
~ 2 times.
Further, material described in step 1) is to pass through pretreated lignocellulosic material, the lignocellulosic
Material can be one of corn stover, wheat stalk, rice straw or wooden fibre known to those skilled in the art
Tie up plain material；It is described pretreatment include the following steps: first by lignocellulosic material using mechanical equipment carry out pulverization process,
So that partial size is reached 0.14 ~ 32 millimeter, is then transmitted by belt conveyor and handled into steam blasting tank, then through plate and frame filter press
Filters pressing makes the water content 50 ~ 65% of material.
Further, the concentration of substrate set described in step 1) is described continuous by material and moisture batch as 20 ~ 25%
It is even to be added in enzymatic vessel, while continuous uniform adds the specific method of lignocellulosic hydrolase and is in batches: object is added for the first time
The weight Zhan of material always adds the 1/8 ~ 1/6 of inventory, and the amount for adding lignocellulosic hydrolase for the first time is always to add the 5/9 of enzyme amount
~ 7/9, when enzymatic vessel completion always adds the 1/2 ~ 3/4 of inventory, remaining 2/9 ~ 4/9 enzyme amount is once added, feed intake end；
Every gram of over dry material adds 30 ~ 60FPU of lignocellulosic hydrolase.
Further, described that material and moisture batch continuous uniform are added in enzymatic vessel in step 1), while in batches
The specific method that continuous uniform adds lignocellulosic hydrolase is: the weight Zhan for adding material for the first time always adds the 1/ of inventory
6, the weight for adding lignocellulosic hydrolase for the first time is always to add the 2/3 of enzyme amount, when enzymatic vessel completion always adds inventory
When 3/4, remaining 1/3 enzyme amount is once added, feed intake end；The end that feeds intake is started to from enzymatic vessel, is completed within 16h, is digested
Step 2 is transferred to when to 48h.
Further, technological condition for fermentation described in step 2 are as follows: fermentation temperature be 30 ~ 32 DEG C, inoculum concentration be 0.5 ~
0.8 ‰, revolving speed is 60 ~ 150r/min.
Further, the process conditions of vacuum distillation described in step 3) are as follows: vapo(u)rizing temperature be 50 ~ 70 DEG C, vacuum degree be-
1.2 ~ -0.90Mpa, distillation time are 0.4 ~ 1.0h, sufficiently finish the ethanolic extraction in fermentation liquid.
Further, the vinasse volume for enzymatic vessel being imported described in step 4) is the 25% of the practical tinning volume of enzymatic vessel.
Compared with prior art, the present invention the beneficial effect is that: the active dry yeasr in fermentation liquid of the present invention is in vacuum distillation
Hot environment under inactivate, in the fermentation liquid in vacuum distillation tower contain non-deactivated cellulase.The present invention is steamed by decompression
It evaporates, remains cellulase activity remaining in fermentation liquid, vinasse is added in new enzymatic hydrolysis system by reuse, it can be with
The lignocellulosic for reducing by 20 ~ 25% hydrolyzes enzyme dosage, the utilization rate of cellulase is greatly improved, to reduce production cost.
Specific embodiment
For a better understanding of the present invention, the contents of the present invention, but this hair are further fairly set out below with reference to embodiment
Bright protection content is not limited solely to the following examples.In the following description, give a large amount of concrete details so as to
More thorough understanding of the invention is provided.It will be apparent, however, to one skilled in the art that the present invention can be with
It is carried out without one or more of these details.In other examples, in order to avoid confusion with the present invention, for
Some technical characteristics well known in the art are not described.
In following embodiments, lignocellulosic hydrolase, also referred to as cellulose complex enzyme or complex cellulase are prepared
Method are as follows: cellulase, zytase, cellobiase are mixed in proportion, 1 ‰ PEG4000 are added, is made.In the present invention,
The filter paper enzyme activity (FPU) of the lignocellulosic hydrolase is 800FPU/mL.Enzyme activity is defined as international unit (IU), i.e. 1min
Enzyme amount needed for generating 1 μm of ol glucose is 1 enzyme-activity unit.Cellulase, zytase, cellobiase, PEG4000 are normal
With reagent, commercially available acquisition can be passed through.
In following embodiments, saccharomyces cerevisiae is provided by Angel Yeast Co., Ltd, and activation method is this field
Routine techniques, method in the present invention are as follows: weigh active dry yeast, the 2%(w/v of sterilized processing is added) glucose is water-soluble
Liquid is under the conditions of 30 DEG C, 30 ~ 45min of aerlbic culture, subject to blistering, can do inoculation and use.
In following embodiments 1 ~ 3, the density of enzymolysis liquid calculates (on-site measurement numerical value) by 1.1kg/L in enzymatic vessel, water
Density is calculated by 1.0kg/L.
Below only by taking wheat stalk as an example, illustrate technical solution of the present invention so that those skilled in the art can implement.
Likewise, those skilled in the art can use other materials, including corn stover, rice straw and other well known wood fibre
One of plain material is a variety of, under the record or enlightenment of the technology of the present invention content, realizes the circulation of enzyme in its enzymolysis process
Utilizing question；It must fall within the scope and spirit of the invention.
Embodiment 1
The present invention will be further described by taking 50L enzymatic vessel as an example.
A kind of method that enzyme recycles in lignocellulosic enzymolysis process, using enzymatic vessel, fermentor and destilling tower string
Connection carries out, and first feeds intake and is digested by the concentration of substrate of setting, and after enzymatic hydrolysis, cooling inoculation is fermented, then by fermentation liquid
It imports in vacuum distillation tower and carries out vacuum distillation, remove ethyl alcohol, active dry yeasr inactivation leaves free cellulase, finally will
Vinasse imports in enzymatic hydrolysis system and carries out reuse, the specific steps are as follows:
1) tap water 19.5L is added into the sky 50L enzymatic vessel M1 that disappears that treated, opens stirring, setting concentration of substrate is equal
It is to be mixed uniform for 25%(w/v), the wheat stalk 3.33kg(1/6 for being 50% (w/w) to M1 investment water content from feeding port)
Afterwards, lignocellulosic hydrolase 0.33L (2/3) is added, controls 50 DEG C of reaction temperature, initially enters enzymolysis process；Then continuous
Remaining lignocellulosic hydrolase 0.17L is added when inlet amount reaches (3/4) 15kg in uniform feeding, continues continuous uniform and throws
Material, feed supplement to total amount reaches 20kg in 16h after starting to feed intake；Every gram of over dry material adds lignocellulosic hydrolase 40FPU；
2) when enzymatic hydrolysis is to 48h, 30 DEG C of inoculation saccharomyces cerevisiaes is cooled to and are fermented, control fermentation condition are as follows: fermentation temperature
It is 30 DEG C, inoculum concentration is 0.5 ‰ (v/v), revolving speed 60r/min；
3) to after fermentation, under 55 ± 2 DEG C, the vacuum degree of -0.95Mpa, carry out vacuum distillation 0.8h to fermentation liquid,
Sufficiently ethanolic extraction therein is finished；
4) 10L concentration of substrate is imported in M1 for the vinasse of 20%(w/v), sequentially adds water 9.5L, lignocellulosic
Hydrolase 0.33L, material 3.4kg control 50 ± 2 DEG C of reaction temperature, initially enter second lot enzymolysis process, specific embodiment party
The same step 1) of method recycles 1 end.
In the present embodiment, wheat stalk is by pretreated material, pretreated method are as follows: first by Wheat Straw
Stalk carries out pulverization process using mechanical equipment, so that partial size is reached 0.14 ~ 32 millimeter, is then transmitted by belt conveyor quick-fried into steam
Broken tank is handled, then makes the water content 50% of material through plate and frame filter press filters pressing.Documented wheat stalk in the following contents
It is identical as the present embodiment, it repeats no more.
It is computed, in cycle period compared with intermittent enzymatic saccharification zymotechnique, lignocellulosic hydrolyzes enzyme dosage
Reduce 20%.
Embodiment 2
With 150m3The present invention will be further described for enzymatic vessel.
A kind of method that enzyme recycles in lignocellulosic enzymolysis process, using enzymatic vessel, fermentor and destilling tower string
Connection carries out, and first feeds intake and is digested by the concentration of substrate of setting, and after enzymatic hydrolysis, cooling inoculation is fermented, then by fermentation liquid
It imports vacuum distillation tower and carries out vacuum distillation, remove ethyl alcohol, active dry yeasr inactivation leaves free cellulase, will finally steam
It evaporates in raffinate importing enzymatic hydrolysis system and carries out reuse, the specific steps are as follows:
1) to the sky 150m that disappears that treated3Tap water 58.5m is added in enzymatic vessel M13, stirring is opened, setting concentration of substrate is equal
For 25%(w/v), from feeding port to 10 tons of wheat stalk (1/6) that M1 investment water content is 50% (w/w), it is to be mixed uniformly after,
Lignocellulosic hydrolase 1.0m is added3(2/3), 50 ± 1 DEG C of reaction temperature are controlled, initially enters enzymolysis process, then continuously
Remaining lignocellulosic hydrolase 0.5m is added when inlet amount is up to 45 tons (3/4) in uniform feeding3, continue continuous uniform and throw
Material, after starting to feed intake in 16h feed supplement to total amount up to 60 tons；Every gram of over dry material adds lignocellulosic hydrolase 40FPU；
2) when enzymatic hydrolysis is to 48h, 32 DEG C of inoculation saccharomyces cerevisiaes is cooled to and are fermented, control fermentation condition are as follows: fermentation temperature
It is 32 DEG C, inoculum concentration is 0.8 ‰ (v/v), revolving speed 150r/min；
3) to after fermentation, under 60 ± 2 DEG C, the vacuum degree of -1.1Mpa, carry out vacuum distillation 0.5h to fermentation liquid,
Sufficiently fraction ethanolic extraction therein is finished；
4) by 30m3Concentration of substrate is 20%(w/v) vinasse import M1 in, sequentially add water 28.5m3, wood fibre
Plain hydrolase 1.0m3, 50 tons of material, control 50 DEG C of reaction temperature, initially enter second lot enzymolysis process, specific implementation method
Same step 1) recycles 1 end.
It is computed, in cycle period compared with intermittent enzymatic saccharification zymotechnique, lignocellulosic hydrolyzes enzyme dosage
Reduce 25%.
Embodiment 3
With 150m3The present invention will be further described for enzymatic vessel.
A kind of method that enzyme recycles in lignocellulosic enzymolysis process, using enzymatic vessel, fermentor and destilling tower string
Connection carries out, and first feeds intake and is digested by the concentration of substrate of setting, and after enzymatic hydrolysis, cooling inoculation is fermented, then by fermentation liquid
It imports vacuum distillation tower and carries out vacuum distillation, remove ethyl alcohol, active dry yeasr inactivation leaves free cellulase, will finally steam
It evaporates in raffinate importing enzymatic hydrolysis system and carries out reuse, the specific steps are as follows:
1) to the sky 150m that disappears that treated3Enzymatic vessel M1 is added 70.8 tons of tap water, opens stirring, and setting concentration of substrate is equal
For 20%(w/v), from feeding port to 8 tons of wheat stalk (1/6) that M1 investment water content is 50% (w/w), it is to be mixed uniformly after, add
Enter lignocellulosic hydrolase 0.8m3(2/3), 50 ± 1 DEG C of reaction temperature are controlled, enzymolysis process is initially entered, it is then continuous equal
Remaining lignocellulosic hydrolase 0.4m is added when inlet amount is up to 36 tons (3/4) in even charging3, continue continuous uniform and throw
Material, after starting to feed intake in 16h feed supplement to total amount up to 48 tons；Every gram of over dry material adds lignocellulosic hydrolase 40FPU；
2) when enzymatic hydrolysis is to 48h, 32 DEG C of inoculation saccharomyces cerevisiaes is cooled to and are fermented, control fermentation condition are as follows: fermentation temperature
It is 32 DEG C, inoculum concentration is 0.6 ‰ (v/v), revolving speed 120r/min；
2) to after fermentation, under 65 ± 2 DEG C, the vacuum degree of -1.2Mpa, carry out vacuum distillation 0.4h to fermentation liquid,
Sufficiently fraction ethanolic extraction therein is finished；
3) by 30m3Concentration of substrate 20%(w/v) vinasse import M1 in, sequentially add water 40.8m3, lignocellulosic water
Solve enzyme 0.8m3, 8 tons of material, control 50 DEG C of reaction temperature, initially enter second lot enzymolysis process, specific implementation method is synchronous
It is rapid 1), recycle 1 time end.
It is computed, in cycle period compared with intermittent enzymatic saccharification zymotechnique, lignocellulosic hydrolyzes enzyme dosage
Reduce 20%.
Claims (4)
1. a kind of method that enzyme recycles in lignocellulosic enzymolysis process, which is characterized in that specific step is as follows:
Step 1): material, moisture batch continuous uniform are added in enzymatic vessel by starting enzymatic vessel according to the concentration of substrate of setting,
Continuous uniform adds lignocellulosic hydrolase in batches simultaneously, is digested；
The concentration of substrate set is described that material and moisture batch continuous uniform are added in enzymatic vessel as 20 ~ 25%, together
When in batches continuous uniform add the specific method of lignocellulosic hydrolase and be: the weight Zhan for adding material for the first time always adds material
The 1/8 ~ 1/6 of amount, the amount for adding lignocellulosic hydrolase for the first time are always to add the 5/9 ~ 7/9 of enzyme amount, initially enter and digest
Journey, then continuous uniform is fed, and when enzymatic vessel completion always adds the 1/2 ~ 3/4 of inventory, is once added remaining 2/9 ~ 4/9
Enzyme amount continues continuous uniform and feeds intake, the end that feeds intake is started to from enzymatic vessel, is completed within 16h, feed intake end；Every gram of over dry object
Material addition 30 ~ 60FPU of lignocellulosic hydrolase,
Step 2: after enzymatic hydrolysis, enzymolysis liquid is cooled to 30 ~ 32 DEG C, saccharomyces cerevisiae is inoculated with, ferments；
Step 3): step 2 fermentation gained fermentation liquid being imported in vacuum distillation tower and carries out vacuum distillation,
The process conditions of the vacuum distillation are as follows: vapo(u)rizing temperature is 50 ~ 70 DEG C, and vacuum degree is -1.2 ~ -0.90Mpa, distillation time
For 0.4 ~ 1.0h, sufficiently the ethanolic extraction in fermentation liquid is finished；
Step 4): vinasse obtained by step 3) vacuum distillation being imported and carries out enzyme reuse in enzymatic vessel, and cycle-index is 1 ~ 2
It is secondary；
The preparation method of the lignocellulosic hydrolase are as follows: mix cellulase, zytase, cellobiase in proportion
It closes, 1 volume, ‰ PEG4000 is added and mixes, is made；The filter paper enzyme activity FPU of the lignocellulosic hydrolase is 800FPU/mL.
2. the method that enzyme recycles in lignocellulosic enzymolysis process as described in claim 1, it is characterised in that: step 1)
The material is to pass through pretreated lignocellulosic material, and the pretreatment includes the following steps: wood fibre first
Plain material carries out pulverization process using mechanical equipment, so that partial size is reached 0.14 ~ 32 millimeter, is then transmitted by belt conveyor into steaming
Vapour cavity charge containers are handled, then make the water content 50 ~ 65% of material through plate and frame filter press filters pressing.
3. the method that enzyme recycles in lignocellulosic enzymolysis process as described in claim 1, it is characterised in that: step 2
Described in technological condition for fermentation are as follows: fermentation temperature be 30 ~ 32 DEG C, inoculum concentration be 0.5 ~ 0.8 ‰, revolving speed be 60 ~ 150r/min.
4. the method that enzyme recycles in lignocellulosic enzymolysis process as described in claim 1, it is characterised in that: step 4)
The vinasse volume for importing enzymatic vessel is the 25% of the practical tinning volume of enzymatic vessel.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974570A (en) * | 2010-09-09 | 2011-02-16 | 天津大学 | Method for recycling cellulose complete components in fuel ethanol production |
| CN104611381A (en) * | 2013-11-05 | 2015-05-13 | 中国石油化工股份有限公司 | Method for producing ethanol by continuous enzymolysis and fermentation of lignocellulose |
| CN104774876A (en) * | 2015-04-10 | 2015-07-15 | 山东龙力生物科技股份有限公司 | Method for comprehensive utilization of lignocellulosic biomasses |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974570A (en) * | 2010-09-09 | 2011-02-16 | 天津大学 | Method for recycling cellulose complete components in fuel ethanol production |
| CN104611381A (en) * | 2013-11-05 | 2015-05-13 | 中国石油化工股份有限公司 | Method for producing ethanol by continuous enzymolysis and fermentation of lignocellulose |
| CN104774876A (en) * | 2015-04-10 | 2015-07-15 | 山东龙力生物科技股份有限公司 | Method for comprehensive utilization of lignocellulosic biomasses |
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