CN104531779A - Method for preparing fuel ethanol by fermenting solid alkali saccharification carbon-enriched micro-algae - Google Patents

Method for preparing fuel ethanol by fermenting solid alkali saccharification carbon-enriched micro-algae Download PDF

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Publication number
CN104531779A
CN104531779A CN201410809268.XA CN201410809268A CN104531779A CN 104531779 A CN104531779 A CN 104531779A CN 201410809268 A CN201410809268 A CN 201410809268A CN 104531779 A CN104531779 A CN 104531779A
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micro
algae
microalgae
solid alkali
fermentation
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袁振宏
周卫征
王忠铭
许敬亮
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Guangzhou Institute of Energy Conversion of CAS
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Guangzhou Institute of Energy Conversion of CAS
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
    • C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • 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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/584—Recycling of catalysts

Abstract

The invention discloses a method for preparing fuel ethanol by fermenting solid alkali saccharification carbon-enriched micro-algae. The method comprises the following steps: by taking micro-algae rich in carbohydrates as raw materials, saccharifying micro-alga dry powder or wet algae under the condition of taking solid alkali as a catalyst, and then performing fermentation to prepare ethanol. Compared with conventional raw materials such as wheat and corns, the micro-algae are easily available and are rapid in growth; and moreover, the content of lignin and hemicellulose in micro-alga cells is low, the content of polysaccharides is rich, and the micro-alga cells contain cellulose Ialpha, hydrogen bonds of the cellulose Ialpha are relatively weak, and thus the cellulose Ialpha is easily degraded into monosaccharide. According to the method disclosed by the invention, the solid alkali catalyst can be recycled and is low in alkalinity, the reaction conditions are easy to control, the method is simple and environmentally friendly, can be used for providing a new route for application of a micro-alga biomass in an aspect of biomass energy, and can be used for exploring a new research direction for the energy utilization research of the micro-algae.

Description

The rich carbon Microalgae fermentation of a kind of solid alkali saccharification prepares the method for alcohol fuel
Technical field:
The present invention relates to technical field prepared by bioenergy, be specifically related to the method that the rich carbon Microalgae fermentation of a kind of solid alkali saccharification prepares alcohol fuel.
Background technology:
Micro-algae, as unicellular or simple many cells photosynthetic microorganism, is distributed widely in various environment, by photosynthesis by sun power, H 2cO in O, air 2be converted into carbohydrate, be stored in microalgae biomass.The growth of micro-algae is with CO in air 2for primary carbon source, often produce 1kg microalgae biomass, the CO of 1.83kg can be fixed 2.For improving the growth velocity of micro-algae, reduce the discharge of greenhouse gases, people utilize the CO in industrial gaseous waste (as stack gas, sodium bicarbonate factory tail gas etc.) usually 2as carbon source.Also have investigator to be combined with wastewater treatment by microdisk electrode, utilize in waste water the feature being rich in the element such as nitrogen, phosphorus, process waste water while production microalgae biomass.From the eighties, under " the water biological species plan " of USDOE is supported, utilizing micro-algae to produce fuel progressively becomes global study hotspot and continues into the present always.In the last few years, the focus that micro-algae diastatic fermentation ethanol becomes each research institution of the world, energy science and technology company is all studied was utilized.Regenerated biological energy seems more and more important in current energy field, and bio-ethanol occupies very important status, nowadays the liquid fuel price of fossil rises steadily, countries in the world all need bio-ethanol to carry out supplementary energy breach, and investigator is devoted to the large-scale production process developing bio-ethanol.At present two maximum U.S. of bio-ethanol producing country and Brazil is in the world all take food crop as raw material.It is raw material with food crop that bio-ethanol all over the world produces great majority, need to take a large amount of arable land, and the consumption of grain also can stimulate food price to go up, and brings many environment and economy problems.And s-generation bio-ethanol, namely cellulosic ethanol is also faced with the obstacle being much difficult to break through.The cheap bio-ethanol of exploitation using non-grain crop as raw material is field of renewable energy topic received much concern.
Micro-algae, compared with traditional raw material preparing bio-ethanol, has that resource is abundanter, the more easy advantage such as acquisition.The nutritious absorption of micro-algae is fast, photosynthetic efficiency is high, grow the features such as rapid.The photosynthetic efficiency of terrestrial plant is general all lower than 0.5%, but the photosynthetic efficiency of micro-algae reaches as high as 10%.Efficient photosynthetic efficiency makes the growth cycle of microalgae cell shorten, its biomass doubling time average out to 2-5 days, and some algae is only 6 hours, can produce a large amount of microalgae biomass at short notice.By manual control condition, both culturing microalgae can carry out the whole year, substantially increases economy, shows that micro-algae is having great advantage as tool in fuel ethyl hydrate raw material.
The raw material that micro-algae prepares ethanol is the carbohydrate in microalgae biomass, mainly comprises starch, Mierocrystalline cellulose, hemicellulose etc.If chlorella, chlamydomonas, grid algae, spirulina etc. are containing a large amount of Mierocrystalline celluloses and starch in micro-algae, there is algae starch content slightly can match in excellence or beauty with other ethanol raw material such as corn, wheat.In addition compared with other lignocellulosic plants, in microalgae cell xylogen and hemicellulose level lower, and different from the cellulose I β in plant, be cellulose I α in microalgae cell, its hydrogen bond is more weak, is more easily degraded to monose.
Summary of the invention:
The object of this invention is to provide the method that the rich carbon Microalgae fermentation of a kind of solid alkali saccharification prepares alcohol fuel.
The present invention is achieved by the following technical programs:
The rich carbon Microalgae fermentation of solid alkali saccharification prepares a method for alcohol fuel, and to be rich in the micro-algae of carbohydrate for raw material, under the condition of solid alkali as catalyzer, by micro-algae dry powder or wet algae saccharification, then fermentation is standby produces ethanol, comprises the following steps:
A, in micro-algae, add water and solid base catalyst, at 60-100 DEG C, be hydrolyzed filtered and recycled solid base catalyst after 60min-180min, by the micro-algae liquid resin absorption process after filtering, then, add 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025gMgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5; The mass ratio of described microalgae dry weight and water is 1:5-1:50; The mass ratio of solid base catalyst and microalgae dry weight is 1:20-1:100;
B, be respectively 20-40FPU/g substrate enzyme amount to the reaction system of step a gained by amylase, cellulase and add amylase, cellulase, 50 DEG C of prehydrolysis 12-24h;
C, be by volume 5% inoculum size yeast (Saccharomyces cerevisiae) is seeded in step b gained reaction system, 30 DEG C fermentation 60-120h, obtain product ethanol.
Described micro-algae is rich in carbohydrate, and carbohydrate content accounts for more than 40% of microalgae dry weight, and starch content accounts for more than 20% of microalgae dry weight.Micro-algae kind that described micro-algae can be cultivated for indoor and outdoor.
Described solid base catalyst is KOH/A1 2o 3, its preparation method is as follows: first by A1 2o 3carrier 500 DEG C of roasting 2-6 hour, adopt equi-volume impregnating by A1 2o 3carrier impregnation is in the KOH solution of 10% in mass concentration, floods 12h under room temperature, dries at terminating latter 110 DEG C, finally in retort furnace in 600 DEG C of roasting 2-6 hours and get final product.
Described yeast, gets the Y2034 bacterial strain of slant preservation, with transfering loop picking one ring yeast thalline, be linked into and be equipped with in the 150mL triangular flask of 50mLYEPD liquid nutrient medium, be positioned in constant-temperature table, regulate temperature at 30 DEG C, shaking culture 12-24 hour under 150rpm condition.Before being inoculated into fermention medium, getting the centrifugal 5min of seed liquor 8000rpm of respective volume, outwell supernatant, inoculation bacterium mud is in fermentation flask.Yeast starter substratum YEPD (w/v): peptone 2%, glucose 2%, yeast powder 1%.
The application compared with prior art, has following beneficial effect:
With the micro-algae of rich starch for raw material, catalyzed by solid base and yeast fermentation is adopted to prepare ethanol, compared with traditional materials of wheat, corn etc., micro-algae is easy to get and grows fast, and in microalgae cell xylogen and hemicellulose level lower, polysaccharide content enrich, and microalgae cell in be cellulose I α, its hydrogen bond is more weak, is more easily degraded to monose.Tradition alkaline purification hydrolysis sugar efficiency is high, but alkali consumption is huge, is difficult to reclaim, and the basicity that carbohydrate hydrolysis simultaneously needs is lower, selects traditional liquid base reaction system stable not, restive.The application's solid base catalyst recoverable, basicity low reaction condition easily controls, simple environmental protection, for the application of microalgae biomass in biomass energy provides new approach, the recent studies on direction for micro-algae recovery energy research and probe.
Embodiment:
Below further illustrate of the present invention, instead of limitation of the present invention.
Embodiment 1:
(1) by chlorella (carbohydrate content 30% ± 2%) algae mud 200g (dry weight 40g) nutrient solution through centrifugal concentrating as in reaction unit, adding volume of water is 400ml, adds solid base catalyst 2g; Temperature of reaction kettle is filtered and recycled solid base catalyst after 60 DEG C of hydrolysis 120min, by the micro-algae liquid resin absorption process after filtration, then, adds 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025g MgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5.
(2) be respectively 40FPU/g substrate enzyme amount to the reaction system of step (1) gained by amylase, cellulase and add amylase and cellulase, 50 DEG C of prehydrolysis 24h.
(3) yeast (YPD liquid nutrient medium 30 DEG C cultivate 24h activation) is seeded in the reaction system that step (2) obtains by the inoculum size of 5% (v/v), 30 DEG C of fermentation 72h, final ethanol 6.12g.
Embodiment 2:
(1) chlorella dry powder 30g (carbohydrate content 30% ± 2%) nutrient solution, as in reaction unit, is added water to 600ml, add solid base catalyst 0.3g.Temperature of reaction kettle is filtered and recycled solid base catalyst after 100 DEG C of hydrolysis 60min, by the micro-algae liquid resin absorption process after filtration, then, adds 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025g MgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5.
(2) be respectively 30FPU/g substrate enzyme amount to the reaction system of step (1) gained by amylase, cellulase and add amylase, cellulase, 50 DEG C of prehydrolysis 18h.
(3) in the reaction system obtained by inoculum size yeast (YPD liquid nutrient medium 30 DEG C cultivate 24h activation) to the step (2) of 5% (v), 30 DEG C of fermentation fermentation 120h, final ethanol 4.26g.
Embodiment 3
(1) will be that micro-plan ball algae dry powder 50g (carbohydrate content 25% ± 2%) nutrient solution, as in high-temperature liquid state water treatment device, adds water 500ml through centrifugal concentrating, add solid base catalyst 2.5g.Temperature of reaction kettle is filtered and recycled solid base catalyst after 80 DEG C of hydrolysis 120min, by the micro-algae liquid resin absorption process after filtration, then, adds 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025g MgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5.
(2) be respectively 20FPU/g substrate enzyme amount to the reaction system of step (1) gained by amylase, cellulase and add amylase, cellulase, 50 DEG C of prehydrolysis 24h.
(3) in the reaction system obtained by inoculum size yeast (YPD liquid nutrient medium 30 DEG C cultivate 24h activation) to the step (2) of 5% (v), 30 DEG C of fermentation fermentation 120h, final ethanol 7.06g.
Embodiment 4:
(1) by chlorella dry powder (carbohydrate content 30% ± 2%) 10g as in reaction unit, adding volume of water is 500ml, adds solid base catalyst 0.25g; Temperature of reaction is filtered and recycled solid base catalyst after 80 DEG C of hydrolysis 90min, by the micro-algae liquid resin absorption process after filtration, then, adds 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025g MgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5.
(2) be respectively 30FPU/g substrate enzyme amount to the reaction system of step (1) gained by amylase, cellulase and add amylase, cellulase, 50 DEG C of prehydrolysis 24h.
(3) yeast (YPD liquid nutrient medium 30 DEG C cultivate 24h activation) is seeded in the reaction system that step (2) obtains by the inoculum size of 5% (v/v), 30 DEG C of fermentation 60h, final ethanol 1.02g.
Embodiment 5:
(1) by chlorella powder 40g nutrient solution as in high-temperature liquid state water treatment device, adding volume of water is 200ml, adds solid base catalyst 0.6g; Temperature of reaction kettle is filtered and recycled solid base catalyst after 70 DEG C of hydrolysis 180min, by the micro-algae liquid resin absorption process after filtration, then, adds 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025g MgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5.
(2) be respectively 30FPU/g substrate enzyme amount to the reaction system of step (1) gained by amylase, cellulase and add amylase, cellulase, 50 DEG C of prehydrolysis 24h.
(3) yeast (YPD liquid nutrient medium 30 DEG C cultivate 24h activation) is seeded in the reaction system that step (2) obtains by the inoculum size of 5% (v/v), 30 DEG C of fermentation 90h, final ethanol 5.89g.

Claims (3)

1. the rich carbon Microalgae fermentation of solid alkali saccharification prepares a method for alcohol fuel, it is characterized in that, to be rich in the micro-algae of carbohydrate for raw material, under the condition of solid alkali as catalyzer, by micro-algae dry powder or wet algae saccharification, then fermentation is standby produces ethanol, comprises the following steps:
A, in micro-algae, add water and solid base catalyst, at 60-100 DEG C, be hydrolyzed filtered and recycled solid base catalyst after 60min-180min, by the micro-algae liquid resin absorption process after filtering, then, add 0.5g (NH by often liter of micro-algae liquid 4) 2hPO 4, 0.025gMgSO 47H 2the amount of O, 1.0g yeast extract paste, adds (NH in the micro-algae liquid after adsorption treatment 4) 2hPO 4, MgSO 47H 2o and yeast extract paste, then add Na 2hPO 4-citrate buffer solution, the pH regulating reaction system is 4.5-5.5; The mass ratio of described microalgae dry weight and water is 1:5-1:50; The mass ratio of solid base catalyst and microalgae dry weight is 1:20-1:100;
B, be respectively 20-40FPU/g substrate enzyme amount to the reaction system of step a gained by amylase, cellulase and add amylase, cellulase, 50 DEG C of prehydrolysis 12-24h;
C, be by volume 5% inoculum size by yeast-inoculated in step b gained reaction system, 30 DEG C fermentation 60-120h, obtain product ethanol.
2. the rich carbon Microalgae fermentation of solid alkali saccharification according to claim 1 prepares the method for alcohol fuel, and it is characterized in that, described solid base catalyst is KOH/A1 2o 3, its preparation method is as follows: first by A1 2o 3carrier 500 DEG C of roasting 2-6 hour, adopt equi-volume impregnating by A1 2o 3carrier impregnation is in the KOH solution of 10% in mass concentration, floods 12h under room temperature, at terminating latter 110 DEG C dry, finally in retort furnace in 600 DEG C of roasting 2-6 hour.
3. the rich carbon Microalgae fermentation of solid alkali saccharification according to claim 1 prepares the method for alcohol fuel, it is characterized in that, described micro-algae is rich in carbohydrate, and carbohydrate content accounts for more than 40% of microalgae dry weight, and starch content accounts for more than 20% of microalgae dry weight.
CN201410809268.XA 2014-12-19 2014-12-19 Method for preparing fuel ethanol by fermenting solid alkali saccharification carbon-enriched micro-algae Pending CN104531779A (en)

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