CN102864188A - Method for producing biodiesel from lignocellulose - Google Patents
Method for producing biodiesel from lignocellulose Download PDFInfo
- Publication number
- CN102864188A CN102864188A CN2011101890933A CN201110189093A CN102864188A CN 102864188 A CN102864188 A CN 102864188A CN 2011101890933 A CN2011101890933 A CN 2011101890933A CN 201110189093 A CN201110189093 A CN 201110189093A CN 102864188 A CN102864188 A CN 102864188A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- vitamin
- algae
- acid
- carbon source
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to the field of microbial fermentation engineering, and particularly relates to a technology of consolidated bioprocessing (consolidated bioprocessing, CBP). Cellulose is fermented and degraded by a cellulose degradation strain (such as clostridium thermal fiber) to generate saccharides and organic acids, such as glucose, lactic acid and acetic acid; and biodiesel and co-produced products with high additional values, such as timnodonic acid, docosahexenoic acid and protein feed, are produced by serial fermentation combined with large-scale heterotrophism or raising cultivation of microalgae (such as schizochytrium limacinum, chlorella and nannochloropsis oculata). The method has the characteristics that anaerobic cellulose degradation bacteria are utilized to degrade the preprocessed lignocellulose material, so that the cellulose and hemicelluloses are converted into carbon sources which can be utilized by the microalgae such as soluble saccharide and organic acids; the carbon sources are inoculated into a microalgae seed liquid to be fermented after a certain nitrogen source and other nutrient elements are added, and oil is accumulated to generate the biodiesel and other products with high additional values.
Description
Technical field
The present invention relates to the microbial fermentation engineering field, specifically utilize and integrate biological processing (Consolidated Bioprocessing, CBP) technology, foundation produces carbohydrate and organic acid such as glucose with cellulose degradation strain (such as Clostridium thermocellum) fermentative degradation Mierocrystalline cellulose, lactic acid, acetic acid etc., unite the extensive heterotrophism of little algae (as splitting kettle algae and little plan ball algae) or foster cultivation of holding concurrently, fermentation to produce biological diesel oil and have high value-added product such as a timnodonic acid (Eicosapentaenoic Acid, EPA) and the cascade fermentation technique of docosahexenoic acid (Docosahexaenoic acid, DHA).
Background technology
Aggravation along with global energy and ecocrisis; seeking substitute energy has become the study hotspot of many countries; this wherein cleans; reproducible bioenergy is the importance of substitute energy research; biofuel is the most suitable product that substitutes petrifaction diesel in the new forms of energy; bring into use as the petrifaction diesel additive in states such as America and Europes, also have certain bottleneck but want mass-producing to use, namely relate to the problem of cost in raw-material selection and the production technique.
Mierocrystalline cellulose is renewable resources the abundantest on the earth, but only has only a few to be used effectively, and the overwhelming majority is used as waste disposal in environment, causes on the contrary certain environmental pollution, such as municipal wastes.So produce novel biological fuel with cellulose substances, cause the extensive attention (Wen Zhiqiang, Jiang Wei, 2010) of countries in the world because having great strategic significance.But lignocellulose causes very difficult (the Christian Weber ﹠amp of its degraded owing to extremely complicated structure; Alexander Farwick et.al, 2010).The Clostridium thermocellum of thermophilic anaerobic (Clostridium thermocellum) is because directly with cellulose conversion being the attention (Lynd.et al, 2002) that monose has attracted numerous investigators.Studies show that (Lamedr, 1984), the cellulosome (cellulosome) that extensively exists in the anaerobic cellulose bacterium for degrading is that it can directly be cellulose conversion the key point of monose, because cellulosome has ribosome-like macromolecular structure, can coordinate, in order, degraded cellulose efficiently.
Little algae has the advantages that as a kind of novel aquatic biomass resource growth cycle is short, yield of biomass is high and fat content is high, and its biomass throughput can reach 30 times of land plant, does not have to reach the huge advantage of not striving grain with the people with grain with striving; Little algae has very attracting great potential at the renewable energy source domain, has very strong grease production ability such as little algae, and its fat content can be up to the 40-80% of dry cell weight.Therefore, little algae might provide an effective solution route for China's Biomass Energy Resources shortage, the little algae resource of develop energy, the little algae biorefinery of development energy industry, the energy strategy demand that not only meets China's Sustainable development, and meet the target that China sets up resource-conserving and friendly environment society, help lend some impetus to harmony between man and nature development and the sustainable development of socio-economy.Utilize heterotrophism mode large-scale culturing micro-algae to have following advantage: (1) micro algae growth reproduction speed is accelerated, and algae biomass concentration improves greatly: when heterotrophism was cultivated, frustule concentration can reach or near the concentration of intestinal bacteria and yeast.(2) from the industrialization angle analysis, the heterotrophism culture systems is more convenient for the control of production process to realize purebred cultivation and stable production.(3) use the heterotrophism culture systems can reduce little algae production cost.
Splitting kettle algae Schizochytrium limacinum is the little algae of a kind of important marine economy, be characterized in that growth is fast, strong stress resistance, lipid content high (reach more than 50% of dry cell weight, and the grease more than 90% existing with the form of triglyceride in the cell).In addition, C14:0 in its lipid acid, C16:0, C22:5 (DPA), C22:6 (DHA) accounts for about 90% of total fatty acid content, has very high nutritive value and relatively easy the separation, therefore can further improve by the by-product high value added product economy of biofuel, split the kettle algae and be considered to one of little algae species of the energy of tool potentiality.
Little plan ball algae (Nannochloropsis sp.) is a kind of single celled marine green algae, has the ability that organic carbons such as utilizing ethanol, acetate and glucose carries out mixotrophic growth.Improve CO in the nutrient solution
2Concentration or add glucose can promote growth and grease productive rate (the Hu ﹠amp of little plan ball frustule; Gao 2003; Xu et al., 2004).The kind of little Sphaerellopsis (Nannochloropsis) has the very fat content of high-content (can reach the 50%-80% of dry cell weight) usually, EPA content can reach the 30%-40% of grease, thereby is considered to be expected to become the Biological resources (Sukenik 1999) of production biofuel and EPA product.
Integrate biological processing technology (Consolidated BioProcessing, be called for short " CBP ") for the lot of challenges that adopts unit operation and integrated optimization pattern to face in the current cellulose utilization technology, integrate by the reactions steps that the scripts such as the production of cellulase, cellulosic enzymic hydrolysis, pentose fermentation and zymohexose is discrete, therefore greatly reduce running cost, thereby improve the complex art economic target of the biological trans-utilization of Mierocrystalline cellulose.Based on these significant characteristics, CBP is considered to one of biological trans-utilization technological line of the most promising low-cost Mierocrystalline cellulose.
It is high that the production of existing biofuel exists cost, strives the problems such as grain with the people.Therefore, seek a kind of low cost, the non-food crop that can utilize again realize that as substrate the production of biofuel is the new problem that the present invention faces.
Summary of the invention
The present invention has set up with anaerobic cellulose bacterium for degrading (conciliating the fiber clostridium such as Clostridium thermocellum) fermentative degradation Mierocrystalline cellulose and has produced carbohydrate and organic acid such as glucose, lactic acid, acetic acid etc., unite the extensive heterotrophism of little algae (as splitting kettle algae and little plan ball algae) or foster cultivation of holding concurrently, fermentation to produce biological diesel oil and polyunsaturated fatty acid such as timnodonic acid (Eicosapentaenoic Acid with high added value, EPA) and the cascade fermentation technique of docosahexenoic acid (Docosahexaenoic acid, DHA).Technical problem to be solved can realize by following scheme:
1. the pre-treatment of lignocellulose (stalk).
Be the calcium hydroxide (Ca (OH) of 20%-30% with stalk and concentration
2) solution is according to 1: 7-1: 15 solid-to-liquid ratio is mixed, and is heated to 90-120 ℃ and keep 100-150min, and processing is cooled to room temperature after finishing, and adding diluted acid, to regulate suspension liquid pH be 7.5-8.0.This mixed solution namely can be used as Clostridium thermocellum (Clostridium thermocellum) or separates fiber clostridium (Clostridium cellulolyticum) degraded cellulose and produce the ferment substrate of required carbon source of follow-up CBP.
2. the required mixed carbon source of Clostridium thermocellum fermentative production downstream fermentation
Clostridium thermocellum high density fermentation culture medium prescription: KH
2PO
41.0-1.5g/L, K
2HPO
44.0-10.0g/L, nitrogenous source 3.0-7.0g/L, MgCl
26H
2O 1.0-5.0g/L, CaCl
22H
2O 50-250mg/L, FeSO
46H
2O 0.5-2.5mg/L, Cysteine hydrochloride 1.0-5.0g/L, Resazurin 1.0-10.0mg/L, carbon source 10.0-40.0g/L, Morpholinopropane sulfonic acid (MOPS) 5.0-20.0g/L, Yeast extract 5.0-20g/L, Sodium citrate2H
2O 0.2-1.5g/L prepares fermention medium, and sterilization.Wherein, carbon source is cellulosic materials biomass (including but not limited to stalks, wooden chrysanthemum), and nitrogenous source is organic or inorganic nitrogenous source (including but not limited to urea, yeast extract, peptone, corn steep liquor, SODIUMNITRATE, Sodium Glutamate); Regulating pH before the sterilization is 7.0-8.5; If need to enlarge concentration of substrate, then must keep carbon-nitrogen ratio is 2: 1-10: 1.
The fermentation condition of Clostridium thermocellum is: temperature 50 C-65 ℃, and anaerobism, tank pressure is 0.05MPa-0.5MPa, mixing speed is 30-100rpm.Should control pH during high concentration of substrate (carbon source concentration is greater than 15g/L) fermentation constant in about 7.0-7.5, fermentation time is 12-48h.
3. the separation of Clostridium thermocellum fermented liquid and little algae utilize the heterotrophic mass culture of CBP series connection
After the Clostridium thermocellum fermentation ends, fermented liquid separates with whizzer, discards solid collection liquid, and sugared content is at 10%-20% in the fermented liquid of separation, add seawater (or artificial seawater) and dilute, the salinity of liquid is between 10-36 after the simultaneously control dilution.Add an amount of nitrogenous source in mixed solution, VITAMIN is mixed with the fermentation culture that nutrient solution carries out little algae.
Chlorella or split the prescription of kettle algae heterotrophic fermentation substratum: carbon source 7%-10%, nitrogenous source 2%-4%: VITMAIN B1,10-50mg/L; Vitamin B6,10-30mg/L; Vitamin B12,1-10mg/L; Vitamin H (vitamin H) 1-10mg/L.Carbon source is produced liquid glucose including but not limited to Clostridium thermocellum, glucose, fructose, wood sugar, lactic acid, glycerine, sodium acetate etc., nitrogenous source be the organic or inorganic nitrogenous source (including but not limited to urea, yeast extract, peptone, corn steep liquor, SODIUMNITRATE, Sodium Glutamate, inorganic ammonium salt); PH value of solution is 5-7.
Chlorella or the fermentation condition that splits the kettle algae are: 23 ℃-28 ℃ of temperature, air flow 0.5-2.0L min
-1L
-1(VVM), tank pressure is 0.04MPa-0.08MPa, and mixing speed is 100-250rpm.Control pH is 5.0-7.5, and fermentation time is 72-96h.
Little plan ball algae is held concurrently and supports the prescription of substratum: carbon source 0.1%-1%; KNO
3, 300-500mg/L; NaH
2PO
42H
2O, 40-70mg/L; FeCl
36H
2O, 6-15mg/L; CuSO
45H
2O, 0.01-0.05mg/L; ZnSO
47H
2O, 0.02-0.08mg/L; CoCl
26H
2O, 0.01-0.05mg/L; MnCl
24H
2O, 0.3-0.5mg/L; NaMoO
42H
2O, 0.01-0.05mg/L; ThiamineHCl, 100-500 μ g/L; Vitamin B12,1-10 μ g/L; Vitamin H (vitamin H) 1-10 μ g/L.Carbon source is produced liquid glucose including but not limited to Clostridium thermocellum, glucose, sodium acetate etc.; PH value of solution is 6-8.
The double hair care ferment condition of little plan ball algae is: 20 ℃-28 ℃ of temperature, air flow 0.1-0.5L min
-1L
-1(VVM), tank pressure is 0.02MPa-0.05MPa, and mixing speed is 30-100rpm, intensity of illumination 50-300 μ mol photons m
-2s
-1Control pH is 5.0-7.5, and incubation time is 8-10 days.
4. according to above step, be that those skilled in the art can realize by lignocellulose production biofuel.The present invention is based on CBP technique thinking, the direct conversion cellulosic materials biomass of exploitation are the technique of biofuel, broken through traditional single spawn culture, adopt lignocellulose degrading bacteria strain (including but not limited to Clostridium thermocellum Clostridium thermocellum JYT01 and Clostridium cellulolyticum H10), with high oil-producing microalgae (including but not limited to splitting kettle Trentepohlia Schizochytrium, little Sphaerellopsis Nannochloropsis, Chlorella Chlorella, chrysophyceae Isochrysis etc.) cascade fermentation, realize conversion accumulation and the high value product of simultaneously attached product of cellulosic degraded and bio-oil, further reduced the production cost of biofuel.Compared with prior art, advantage of the present invention and positively effect are:
5. utilize lignocellulose degrading bacteria strain degraded through pretreated cellulosic material, make Mierocrystalline cellulose wherein change the carbon source material that little algaes such as carbohydrate and organic acid can utilize into, after utilizing this carbon source and adding certain nitrogenous source and other nutritive element, access little algae seed liquor and carry out CBP associating heterotrophism (or hold concurrently and support) fermentation, accumulation grease production biofuel and other high value added product.
Description of drawings
Nothing
Embodiment
The present invention is further detailed explanation below in conjunction with embodiment.
Be described in detail for following problem below in conjunction with concrete experiment:
Determining of the pretreatment technology of I, ligno-cellulosic materials;
Growing state when II, Clostridium thermocellum lignocellulose degradation;
III, CBP unite the growing state that splits the kettle algae when splitting kettle algae heterotrophic fermentation;
IV, CBP unite the growing state of little plan ball algae little plan ball algae when holding concurrently the hair care ferment.
Embodiment 1: the stalk Pretreatment Test
1. stalk is rolled into segment, and be ground into 40 purpose powder with pulverizer.
2. take by weighing the 1.5g straw powder to tube sealing, (mass ratio) adds NaOH and Ca (OH) in proportion
2, and add entry according to solid-to-liquid ratio.
3.100 ℃ processing 2h.
4. transfer pH to neutral with sulfuric acid after the cooling, and be filtered dry a pot suction filtration with crossing.Water cleans several times behind the suction filtration, abandons filtrate.Do pot and process after 80 ℃ of straw powder dry to constant weight.
5. take by weighing straw powder 100mg after the processing in the enzymolysis bottle, add 2mL 0.05mol/L pH=5.47 Citric acid buffer.Press the 1g solid and add 25U ctec (cellulase of Novi's letter), 50 ℃ of enzymolysis 24h.Blank is not add substrate among the 2mL 0.05mol/L pH=5.47 Citric acid buffer, only adds the cellulase of equivalent.
6.48h after, all substances in the enzymolysis bottle are transferred in the volumetric flask, constant volume extracts part solution, with 0.2 μ m membrane filtration.HPLC detects to get sugared rate (table 1).
The effect of table 1. different pretreatments (by must be sugared rate calculate)
Process 1:1%NaOH; Process 2:27%Ca (OH)
2Process 3:1%NaOH+10%Ca (OH)
2
Process 4:1%NaOH+27%Ca (OH)
2Process 5: contrast (filter paper); Process 6: contrast (Mierocrystalline cellulose)
According to experimental result, consider and treatment effect and Financial cost, final determine to process 2, namely utilizing mass ratio is 27% Ca (OH)
2The effect of Treating straw is best.
Embodiment 2:5L ferment tank Clostridium thermocellum
1. by prescription " KH
2PO
41.0g/L, K
2HPO
4(anhydrous) 6g/L, urea 17.8g/L, MgCl
26H
2O 2.49g/L, CaCl
22H
2O 135mg/L, FeSO
46H
2O 5mg/L, Cysteine hydrochloride 2.0g/L, Resazurin 2.0mg/L, cellobiose 30.0g/L, Yeast extract 12.0g/L, Sodium citrate2H
2O 0.752g/L " preparation fermention medium (adding after calcium, magnesium, the mother liquid of iron salt sterilization), 115 ℃, the 15min moist heat sterilization.
2. after sterilization finishes, constantly in fermentor tank, pass into nitrogen, to remove the oxygen in substratum and the fermentor tank.After substratum became the color of himself again by light blue or pink (color of resazurin), regulating medium pH constant was 7.0, and 60 ℃ of tank temperature access volume required JYT01 primary seed solution simultaneously, stopped ventilation, and 80rpm stirs fermentation.
Approximately behind the 48h carbon source all consumed, cell concentration reaches 12.7 (OD in the fermented liquid
600).
Embodiment 3:CBP unites and splits kettle algae heterotrophic fermentation production biofuel
Adopt above embodiment gained Clostridium thermocellum to produce liquid glucose (carbon source content 10%), other adds yeast extract 3%, VITMAIN B1,40mg/L; Vitamin B6,20mg/L; Vitamin B12,10mg/L; Vitamin H (vitamin H) 10mg/L.Carry out fermentation culture by following condition: 25 ℃ of temperature, air flow 1.0L min-1 L-1 (VVM), tank pressure are 0.06MPa, mixing speed is 200rpm.Control pH is 6.0, and fermentation time is 96h.
After the fermentation ends, the output that the kettle algae is split in the acquisition of 5L fermentor tank is 44.15g/L, obtains bio-oil 23.8g/L, and DHA content is 9g/L (table 2,3).
Table 2 CBP combined ferment obtains to split the composition analysis result of kettle algae
Table 3 CBP fermentation obtains to split the fatty acid analysis result of kettle algae
Embodiment 4:CBP unites little plan ball algae hair care ferment production biofuel of holding concurrently
Little plan ball algae is held concurrently and supports the prescription of substratum: Clostridium thermocellum produces liquid glucose (carbon source content) 0.6%, KNO
3, 400mg/L; NaH
2PO
42H
2O, 60mg/L; FeCl
36H
2O, 10mg/L; CuSO
45H
2O, 0.02mg/L; ZnSO
47H
2O, 0.05mg/L; CoCl
26H
2O, 0.02mg/L; MnCl
24H
2O, 0.4mg/L; NaMoO
42H
2O, 0.02mg/L; ThiamineHCl, 200 μ g/L; Vitamin B12,5 μ g/L; Vitamin H (vitamin H) 5 μ g/L.
The double hair care ferment condition of little plan ball algae is: 22 ℃ of temperature, and air flow 0.2L min-1 L-1 (VVM), tank pressure are 0.02MPa, mixing speed is 60rpm, intensity of illumination 300 μ mol photons m
-2s
-1Control pH is 7.2, and incubation time is 10 days.
After the fermentation ends, the output that the 5L fermentor tank obtains little plan ball algae is 2.73g/L, obtains bio-oil 1.49g/L, and EPA content is 226mg/L (table 4).
Table 4 CBP fermentation obtains the fatty acid analysis result of little plan ball algae
More than be sub to further describing that the present invention does in conjunction with specific embodiments.The technician of the industry should understand; the present invention is not restricted to the described embodiments; that describes in above-described embodiment and the specification sheets just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.
Claims (8)
1. the method for a lignocellulose production biofuel, it is characterized in that utilizing the biological processing technology (CBP) of integrating, produce carbohydrate and organic acid such as glucose with lignocellulose degrading bacteria strain degraded cellulose, lactic acid, acetic acid etc., unite the extensive heterotrophism of little algae or hold concurrently to support cultivate, fermentation to produce biological diesel oil and have the cascade fermentation technique of the polyunsaturated fatty acid of high added value may further comprise the steps:
1) pre-treatment of lignocellulose (stalk)
With stalk and calcium hydroxide (Ca (OH)
2) solution is according to 1: 7-1: 15 solid-to-liquid ratio is mixed, and is heated to 90-120 ℃ and keep 60-150min, and processing is cooled to room temperature after finishing, and adding diluted acid, to regulate suspension liquid pH be 7.5-8.0;
2) the required mixed carbon source of degraded cellulose fermentative production downstream fermentation
Clostridium thermocellum high density fermentation culture medium prescription: KH
2PO
41.0-1.5g/L, K
2HPO
44.0-10.0g/L, nitrogenous source 3.0-7.0g/L, MgCl
26H
2O 1.0-5.0g/L, CaCl
22H
2O 50-250mg/L, FeSO
46H
2O 0.5-2.5mg/L, Cysteine hydrochloride 1.0-5.0g/L, Resazurin 1.0-10.0mg/L, carbon source 10.0-40.0g/L, Morpholinopropane sulfonic acid (MOPS) 5.0-20.0g/L, Yeast extract 5.0-20g/L, Sodium citrate2H
2O 0.2-1.5g/L prepares fermention medium, and sterilization;
Fermentation condition is: temperature 50 C-65 ℃, and anaerobism, tank pressure is 0.1MPa-0.5MPa, and mixing speed is 30-100rpm, and pH is 7.0-7.5, and fermentation time is 12-48h;
3) large-scale culturing micro-algae that utilizes the CBP technology to connect
After the Clostridium thermocellum fermentation ends, fermented liquid separates with whizzer, discards solid collection liquid, and sugared content is at 10%-20% in the fermented liquid of separation, add seawater (or artificial seawater) and dilute, the salinity of liquid is between 10-36 after the simultaneously control dilution.Add an amount of nitrogenous source in mixed solution, VITAMIN is mixed with the fermentation culture that nutrient solution carries out little algae; Chlorella or split the prescription of kettle algae heterotrophic fermentation substratum: carbon source 7%-10%, nitrogenous source 2%-4%, VITMAIN B1,10-50mg/L; Vitamin B6,10-30mg/L; Vitamin B12,1-10mg/L; Vitamin H (vitamin H) 1-10mg/L;
Chlorella or the fermentation condition that splits the kettle algae are: 23 ℃-28 ℃ of temperature, air flow 0.5-2.0L min
-1L
-1, tank pressure is 0.04MPa-0.08MPa, and mixing speed is 100-250rpm, and control pH is 5.0-7.5, and fermentation time is 72-96h;
The prescription of little plan ball algae culture medium: carbon source 0.1%-1%, KNO
3300-500mg/L; NaH
2PO
42H
2O 40-70mg/L; FeCl
36H
2O 6-15mg/L; CuSO
45H
2O 0.01-0.05mg/L; ZnSO
47H
2O 0.02-0.08mg/L; CoCl
26H
2O 0.01-0.05mg/L; MnCl
24H
2O 0.3-0.5mg/L; NaMoO
42H
2O 0.01-0.05mg/L; ThiamineHCl 100-500 μ g/L; Vitamin B12,1-10 μ g/L; Vitamin H (vitamin H) 1-10 μ g/L;
The double hair care ferment condition of little plan ball algae is: 20 ℃-28 ℃ of temperature, air flow 0.1-0.5L min
-1L
-1, tank pressure is 0.02MPa-0.05MPa, mixing speed is 30-100rpm, intensity of illumination 50-300 μ mol photons m
-2s
-1, control pH is 5.0-7.5, incubation time is 8-10 days.
2. method according to claim 1, the concentration of preferably calcium hydroxide is 20%-30% during pre-treatment, preferred Heating temperature is 100 ℃-110 ℃.
3. method according to claim 1, when the Clostridium thermocellum high density fermentation is cultivated, carbon source is cellulosic materials biomass (including but not limited to stalks, wooden chrysanthemum), and nitrogenous source is that the organic or inorganic nitrogenous source is (including but not limited to urea, yeast extract, peptone, corn steep liquor, SODIUMNITRATE, Sodium Glutamate, inorganic ammonium salt); If need to enlarge concentration of substrate, then must keep carbon-nitrogen ratio is 2: 1-10: 1.
4. method according to claim 1, when the Clostridium thermocellum high density fermentation was cultivated, optimum condition was: 58 ℃-62 ℃ of temperature, anaerobism, mixing speed is 30-70rpm, fermentation time is 24-48h;
5. method according to claim 1, CBP cascade fermentation chlorella or when splitting the kettle algae, carbon source is produced liquid glucose including but not limited to Clostridium thermocellum, glucose, fructose, wood sugar, lactic acid, glycerine, sodium acetate etc., nitrogenous source be the organic or inorganic nitrogenous source including but not limited to urea, yeast extract, peptone, corn steep liquor, SODIUMNITRATE, Sodium Glutamate, inorganic ammonium salt; When CBP connected the little plan ball of facultative cultivation algae, carbon source was produced liquid glucose including but not limited to Clostridium thermocellum, glucose, sodium acetate etc.
6. method according to claim 1, CBP cascade fermentation chlorella or when splitting the kettle algae, screening formulation is: carbon source 7%-9%, nitrogenous source 2%-3%, VITMAIN B1,30-50mg/L; Vitamin B6,10-20mg/L; Vitamin B12,5-10mg/L; Vitamin H 5-10mg/L; When CBP connected the little plan ball of facultative cultivation algae, screening formulation was: carbon source 0.3%-0.6%; KNO
3400-450mg/L; NaH
2PO
42H
2O 40-60mg/L; FeCl
36H
2O 8-12mg/L; CuSO
45H
2O 0.01-0.03mg/L; ZnSO
47H
2O 0.04-0.05mg/L; CoCl
26H
2O 0.01-0.02mg/L; MnCl
24H
2O 0.3-0.4mg/L; NaMoO
42H
2O 0.01-0.03mg/L; ThiamineHCl 150-250 μ g/L; Vitamin B12,1-5 μ g/L; Vitamin H 1-5 μ g/L; Little plan ball algae preferably double hair care ferment condition is: 20 ℃-23 ℃ of temperature, air flow 0.1-0.3L min
-1L
-1, mixing speed is 30-70rpm, intensity of illumination 70-200 μ mol photons m
-2s
-1
7. method according to claim 1, it is characterized in that used lignocellulose degrading bacteria strain is including but not limited to Clostridium thermocellum Clostridium thermocellum JYT01 and Clostridium cellulolyticum H10, high oil-producing microalgae is including but not limited to splitting kettle Trentepohlia Schizochytrium, little Sphaerellopsis Nannochloropsis, Chlorella Chlorella, chrysophyceae Isochrysis, and any mutant strain that carries out on their bases etc.
8. method according to claim 1 is characterized in that the mode batch fermentation of described fermentation, and feed supplement-batch fermentation continuously ferments or semicontinuous fermentation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101890933A CN102864188A (en) | 2011-07-07 | 2011-07-07 | Method for producing biodiesel from lignocellulose |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101890933A CN102864188A (en) | 2011-07-07 | 2011-07-07 | Method for producing biodiesel from lignocellulose |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102864188A true CN102864188A (en) | 2013-01-09 |
Family
ID=47443315
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011101890933A Pending CN102864188A (en) | 2011-07-07 | 2011-07-07 | Method for producing biodiesel from lignocellulose |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102864188A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104388317A (en) * | 2014-12-03 | 2015-03-04 | 东莞市绿安奇生物工程有限公司 | Production method of organic chlorella enriched in trace elements |
| CN106367129A (en) * | 2016-08-30 | 2017-02-01 | 南宁华侨投资区政孙贸易有限公司 | Method for producing biodiesel from microalgae |
| CN106916856A (en) * | 2015-12-28 | 2017-07-04 | 丰益(上海)生物技术研发中心有限公司 | Improve the culture medium and method of lipid-producing microorganisms production odd-carbon fatty acid yield |
| CN108977470A (en) * | 2018-08-17 | 2018-12-11 | 中国科学院青岛生物能源与过程研究所 | The method that polyunsaturated fatty acid grease is rich in using lignocellulosic production |
| CN108977401A (en) * | 2018-08-17 | 2018-12-11 | 中国科学院青岛生物能源与过程研究所 | Using the method for lignocellulosic culture microalgae |
| CN109097416A (en) * | 2018-08-17 | 2018-12-28 | 中国科学院青岛生物能源与过程研究所 | Lignocellulosic one kettle way bioconversion method |
| CN109221620A (en) * | 2018-10-30 | 2019-01-18 | 中国科学院青岛生物能源与过程研究所 | A kind of lignocellulosic base biological feedstuff and preparation method |
| CN109380597A (en) * | 2018-10-30 | 2019-02-26 | 中国科学院青岛生物能源与过程研究所 | A kind of preparation method of nonreactive biology chicken feed |
| CN110771424A (en) * | 2019-09-25 | 2020-02-11 | 郭红伟 | Phellinus igniarius cultivation method for accumulating anti-tumor effective ingredients |
| CN113881710A (en) * | 2021-10-29 | 2022-01-04 | 安徽工程大学 | Method for co-producing biological hydrogen and microalgae grease by utilizing lignocellulose fermentation |
-
2011
- 2011-07-07 CN CN2011101890933A patent/CN102864188A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| 《生物化学与生物物理进展》 20111231 王金兰等 降解纤维素的"超分子机器"研究进展 28-35 1-8 第38卷, 第1期 * |
| MATTHEW N CAMPBELL: "Biodiesel:Algae as a Renewable Source for Liquid Fuel", 《GUELPH ENGINEERING JOURNAL》 * |
| 李臣: "微藻生物柴油研究进展", 《安徽农业科学》 * |
| 王金兰等: "降解纤维素的"超分子机器"研究进展", 《生物化学与生物物理进展》 * |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104388317A (en) * | 2014-12-03 | 2015-03-04 | 东莞市绿安奇生物工程有限公司 | Production method of organic chlorella enriched in trace elements |
| CN106916856A (en) * | 2015-12-28 | 2017-07-04 | 丰益(上海)生物技术研发中心有限公司 | Improve the culture medium and method of lipid-producing microorganisms production odd-carbon fatty acid yield |
| CN106367129A (en) * | 2016-08-30 | 2017-02-01 | 南宁华侨投资区政孙贸易有限公司 | Method for producing biodiesel from microalgae |
| CN109097416B (en) * | 2018-08-17 | 2020-10-20 | 中国科学院青岛生物能源与过程研究所 | Lignocellulose one-pot biotransformation method |
| CN108977401A (en) * | 2018-08-17 | 2018-12-11 | 中国科学院青岛生物能源与过程研究所 | Using the method for lignocellulosic culture microalgae |
| CN109097416A (en) * | 2018-08-17 | 2018-12-28 | 中国科学院青岛生物能源与过程研究所 | Lignocellulosic one kettle way bioconversion method |
| CN108977470A (en) * | 2018-08-17 | 2018-12-11 | 中国科学院青岛生物能源与过程研究所 | The method that polyunsaturated fatty acid grease is rich in using lignocellulosic production |
| CN108977401B (en) * | 2018-08-17 | 2021-09-28 | 中国科学院青岛生物能源与过程研究所 | Method for culturing microalgae by adopting lignocellulose |
| CN108977470B (en) * | 2018-08-17 | 2022-04-15 | 中国科学院青岛生物能源与过程研究所 | Method for producing polyunsaturated fatty acid-rich oil by adopting lignocellulose |
| CN109221620A (en) * | 2018-10-30 | 2019-01-18 | 中国科学院青岛生物能源与过程研究所 | A kind of lignocellulosic base biological feedstuff and preparation method |
| CN109380597A (en) * | 2018-10-30 | 2019-02-26 | 中国科学院青岛生物能源与过程研究所 | A kind of preparation method of nonreactive biology chicken feed |
| CN110771424A (en) * | 2019-09-25 | 2020-02-11 | 郭红伟 | Phellinus igniarius cultivation method for accumulating anti-tumor effective ingredients |
| CN110771424B (en) * | 2019-09-25 | 2022-01-07 | 宁波御菌生物技术有限公司 | Phellinus igniarius cultivation method for accumulating anti-tumor effective ingredients |
| CN113881710A (en) * | 2021-10-29 | 2022-01-04 | 安徽工程大学 | Method for co-producing biological hydrogen and microalgae grease by utilizing lignocellulose fermentation |
| CN113881710B (en) * | 2021-10-29 | 2023-09-22 | 安徽工程大学 | Method for co-production of biological hydrogen and microalgae grease by lignocellulose fermentation |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102864188A (en) | Method for producing biodiesel from lignocellulose | |
| CN102888348B (en) | Schizochytrium limacinum and method or fermenting and producing DHA (Docosahexaenoic Acid) grease utilizing high density of schizochytrium limacinum | |
| CN101735993B (en) | Method for efficiently producing cellulase | |
| CN102174433B (en) | Clostridium beijerinckii with high stress resistance and application thereof | |
| CN105039437A (en) | Method using molasses, alcohol, and waste undecanted wine to culture monoraphidium to produce diesel oil | |
| CN103993042A (en) | Method for combined production of bioethanol and pullulan from lignocellulose substances | |
| CN113308387B (en) | Bacterial strain for co-production of unsaturated fatty acid and carotenoid and application thereof | |
| CN104031843A (en) | Aurantiochytrium sp. and application | |
| WO2021254076A1 (en) | Oleaginous yeast and use thereof | |
| CN103937843A (en) | Method for producing DHA (Docosahexaenoic Acid) by fermenting schizochytrium limacinum via mixed carbon source | |
| CN109097417A (en) | Improve the full bacterium method for saccharifying of lignocellulosic saccharification efficiency | |
| CN102417915A (en) | Method for producing microbial grease by fermenting inulin serving as raw material | |
| CN103421850A (en) | Method used for producing bioethanol with Scenedesmusabundans | |
| CN104004794B (en) | A kind of method utilizing cellulose mixture enzyme crude enzyme liquid hydrolysis of lignocellulose fermentation to prepare n-butyl alcohol | |
| CN104212846A (en) | Method for producing DHA (docosahexenoic acid) by using cheap bean pulp as nitrogen source for fermenting schizochytrium | |
| CN105062928B (en) | A kind of zymomonas mobilis and its application of resisting high-concentration acetic acid and high concentration furtural | |
| CN103074380B (en) | Method for improving anaerobic energy production efficiency of algae raw material through bacterial degradation biological pretreatment | |
| CN111411141B (en) | Method for producing microbial grease by co-fermenting corn straws with bacterial enzymes | |
| CN101497871B (en) | Alcohol fermentation anaerobic high temperature bacterium culture medium, preparation and use thereof | |
| CN108103117A (en) | A kind of method using saccharomycetes to make fermentation corncob production biodiesel and its manufactured biodiesel | |
| CN103436567A (en) | Method for producing kojic acid by using molasses fermented by immobilized aspergillus oryzae | |
| CN113881710B (en) | Method for co-production of biological hydrogen and microalgae grease by lignocellulose fermentation | |
| CN103045491A (en) | Mixed culture method of microorganism containing oil | |
| CN103773694B (en) | A kind of method of high-efficient culture oil-containing microorganism | |
| CN103290068B (en) | Method for producing erythritol by utilizing hypertonicity-resistant Aureobasidium pullulans |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20130109 |