CN104805137A - Method for producing gluconic acid through bioconversion of lignocellulose - Google Patents
Method for producing gluconic acid through bioconversion of lignocellulose Download PDFInfo
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- CN104805137A CN104805137A CN201410034928.1A CN201410034928A CN104805137A CN 104805137 A CN104805137 A CN 104805137A CN 201410034928 A CN201410034928 A CN 201410034928A CN 104805137 A CN104805137 A CN 104805137A
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- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
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Abstract
The present invention relates to a method for efficiently producing gluconic acid through bioconversion of lignocellulose by using Aspergillus niger. The method specifically comprises: inoculating cultured Aspergillus niger in a lignocellulose hydrolyzate to ferment so as to produce gluconic acid, wherein the gluconic acid concentration can achieve 80 g/L after fermentation for 96 h; or simultaneously adding Aspergillus niger being subjected to liquid culture, a lignocelluloses raw material, and cellulase into a reactor, and carrying out synchronous saccharification and fermentation so as to produce gluconic acid. According to the present invention, the method has characteristics of short time and high efficiency, the lignocellulose has the wide source and the low price, and the broad development space and the broad development prospects are provided.
Description
[technical field]
The present invention relates to biology manufacture and the biorefinery field of bio-based chemical, specifically, is the lignocellulosic materials such as maize straw through the method for saccharification and the acid of fermentative production high concentration glucose.
[background technology]
Gluconic acid and its esters have nontoxic, Heat stability is good and the not easily feature such as deliquescence, have been widely used in industries such as chemical industry, food, medicine, light industrys.Particularly in cement additive, Sunmorl N 60S can be widely used as water reducer and retardant, and achieves good effect; Can be used as accessory substance, solidifying agent, buffer reagent etc. in foodstuffs industry, present people have been the condiment that development of raw materials goes out high flavor with Sunmorl N 60S.In addition, water conditioning, plating, metal and the multiple industrial sector such as nonmetallic surface cleaning and manufacture of cement is also widely used in because gluconic acid has excellent chelating ability.At present, the annual requirement of gluconic acid is more than 500,000 tons.
The method of producing gluconic acid mainly contains biological fermentation process, eletrooxidation method, electrolytic oxidation and heterogeneous catalytic oxidation method four kinds of methods.The intermediate steps of eletrooxidation method is many, and by product is many, and product is difficult to be separated, and containing sodium-chlor in finished product, and its application in cement additive is restricted; Electrolytic oxidation aborning energy consumption is large and wayward; Heterogeneous catalytic oxidation method needs to use expensive catalyzer, causes the production cost of gluconic acid to remain high.And microbe fermentation method is due to advantages such as fermenting speed is fast, fermenting process is easy to control, product easily extracts, administration measure gentle, easy suitability for industrialized production, and become the topmost production method of glucose present acid.
Raw material mainly glucose, molasses and the starchy material of fermentative production gluconic acid.And current gluconic acid output can not meet the demand to gluconic acid day by day increased, the extensive simply enlarging of starch deep processing enterprise will certainly badly influence the grain security of country.Therefore, seek cheap and the gluconic acid raw materials for production of wide material sources are matters of utmost importance that gluconic acid industry healthy development faces.China is large agricultural country, and the output of the lignocellulosic materials such as agroforestry waste is huge, but major part is all slatterned by the burning of poor efficiency, not only causes the huge wasting of resources, and result in more serious environment and traffic problems.And utilize lignocellulosic material to produce gluconic acid, then effectively can solve the raw material sources problem of gluconic acid industry, effectively can alleviate current rural economy, rural development and rural demography simultaneously, the deep processing for agricultural wastes provides new outlet.
Bio-transformation lignocellulosic material production of chemicals, generally comprises the operations such as raw materials pretreatment, enzymic hydrolysis, fermentation and product separation.Lignocellulosic material destroys its dense structure through pre-treatment, promotes that cellulase is to cellulosic degraded.But preprocessing process produces the inhibitions such as organic acid, aldehydes and phenols.In order to improve the fermentability of cellulose matrix, must reduce before fermentation or removing inhibition wherein completely.But being that substrate fermentation production gluconic acid exists many technological difficulties with lignocellulose, mostly being heterogeneous catalytic oxidation method about utilizing lignocellulosic material to produce the method adopted in the report of gluconic acid at present.Adopt these methods to produce in the process of gluconic acid, need to use the chemical such as a large amount of sulfuric acid, sodium hydroxide, and catalyzer mostly is Na
2cu (OH)
4with gold/phosphorus heteropoly tungstic acid cesium salt, although this method transformation efficiency is high, used catalyst is limited in cycle index, and the catalyst consumption amount of unit product is high, in a disguised form adds the production cost of gluconic acid product.
The invention provides with completely real lignocellulosic material for matrix, the pre-treatment adopting contriver to propose, saccharification and zymotechnique produce high concentration glucose acid.
[summary of the invention]
The object of the invention is the fermented substrate expanding crude dextrose acid-fermentation, uses cheap, resourceful lignocellulose, and providing a kind of is raw material with lignocellulose, by the method for saccharification and the acid of fermentative production high concentration glucose.
Design of the present invention: be that raw material low-cost high-efficiency produces gluconic acid with lignocellulose, specifically first carry out the pre-treatment of lignocellulosic material, the raw material obtained carries out detoxification or not detoxification, then produce gluconic acid through following two kinds of approach: one, obtain saccharified liquid with the raw material after cellulase degradation process, then access gluconic acid production bacterial strain and carry out gluconic acid fermentation; Two, raw material, cellulase and the gluconic acid after process is produced bacterial strain to add in reactor simultaneously, produce gluconic acid by the mode of synchronous saccharification and fermentation.
The object of the invention is to be achieved through the following technical solutions:
Bio-transformation lignocellulose produces a method for gluconic acid, and it is characterized in that, its concrete steps are:
(1) lignocellulose pre-treatment:
Lignocellulose pre-treatment comprises dilute acid pretreatment, steam explosion, and swollen quick-fried, the sulphite pre-treatment of the explosion of ammonia fiber, liquefied ammonia, alkaline process pre-treatment, machinery grind pre-treatment, Biological Pretreatment; Preferred dilute acid pretreatment method, dilute acid pretreatment refers to that dilution heat of sulfuric acid infiltrates material, then at 180 ~ 200 DEG C, maintains 1 ~ 10min, destroys lignocellulose structure, thus is conducive to follow-up enzymolysis process.
Described lignocellulosic material comprises one or more the mixing such as maize straw, wheat-straw, straw, Cotton Stalk, energy-source plant, wood, wood chip, waste paper, agroforestry waste.
Described lignocellulosic material have passed through pre-treatment, its dense structure is destroyed, thus improves its enzymolysis.Pretreatment process includes but not limited to that swollen quick-fried, the sulphite pre-treatment of dilute acid pretreatment, steam explosion, the explosion of ammonia fiber, liquefied ammonia, alkaline process pre-treatment, machinery grind pre-treatment, Biological Pretreatment etc., preferred dilute acid pretreatment method.
(2) liquid nutrient medium is utilized to cultivate aspergillus niger:
40 ~ 80g/L glucose, 0 ~ 1g/L magnesium sulfate, 0 ~ 2g/L SODIUM PHOSPHATE, MONOBASIC, the potassium primary phosphate of 0 ~ 2g/L, the manganous sulfate of 0 ~ 2g/L, liquid culture aspergillus niger 25 ~ 40 DEG C, 50 ~ 500rpm, cultivation is 12 ~ 48h, and cell concentration reaches 1 ~ 3g/L.The aspergillus niger cultivated is used for further cultivation or the fermentation of liquid or solid fermented substrate.
(3) solid-state biological detoxication:
After regulating lignocellulosic material pH and water content, access detoxification mould Amorphotheca resinaeZN1, deposit number CGMCC7452, solid state rheology is characterised in that culture temperature 20 ~ 35 DEG C, and material moisture is 30 ~ 70%, material pH4.5 ~ 6.5.The compound that detoxification removes comprises formic acid, acetic acid, furfural, hydroxymethylfurfural etc.
If the gluconic acid selecting inhibition tolerance strong produces bacterium, then step (3), namely can omit the detoxification treatment of lignocellulosic material.
If bacterium produced by the gluconic acid selecting inhibition tolerance poor, then pretreated lignocellulosic material need through detoxification treatment, remove the inhibition produced in preprocessing process, comprise organic acid compounds, as formic acid, acetic acid, levulinic acid etc., furan derivative, as furfural, hydroxymethylfurfural etc., phenolic compound, as p-Hydroxybenzaldehyde, vanillin food grade,1000.000000ine mesh etc.Poison-removing method includes but not limited to biological detoxication, washing, crosses alkaline purification (Overliming), charcoal absorption, ionic adsorption etc., preferred biological detoxication.
(4) production of gluconic acid:
Add lignocellulosic material and the cellulase of detoxification or non-detoxification, water use regulation solids content, at 50 DEG C in fermentor tank simultaneously, saccharification 48 ~ 96h under the condition of pH4.8, solid-liquid separation is carried out to the slurries after saccharification, obtains supernatant liquor, i.e. lignocellulose enzymolysis liquid; The aspergillus niger strain that liquid culture is good, be inoculated in hydrolyzed solution and ferment.
Or the aspergillus niger strain, lignocellulosic material (detoxification or non-detoxification) and the cellulase that liquid culture are obtained join in reactor simultaneously, at 30 ~ 50 DEG C, synchronous saccharification and fermenting lignocellulose raw material production gluconic acid under the condition of pH4 ~ 6, fermentation time is 72 ~ 120h.
The massfraction of described lignocellulose in fermentor tank is 10 ~ 40%, and cellulase is 5 ~ 30FPU/g DM, leavening temperature 30 ~ 50 DEG C.Described cellulose raw material is selected from solid straws raw material or solid straws handled thing; Wherein: described cellulose raw material comprises in maize straw, wheat straw, straw, wood chip, energy-source plant or forestry waste.
The bacterial strain producing grape acid mainly contains aspergillus niger, mould, glucose oxidation and bacillus etc., and wherein aspergillus niger is more suitable for utilizing lignocellulosic material to produce gluconic acid.
The gluconic acid that fermentation generates, comprises the gluconic acid of free state, and the gluconate regulating fermentation pH to generate, as Sunmorl N 60S, and calglucon, ammonium gluconate etc.
Compared with prior art, positively effect of the present invention is:
(1) the present invention is fermenting raw materials gluconic acid with lignocellulose, and matrix is with low cost, wide material sources, takes full advantage of renewable resources, does not strive grain with the mankind, can meet the market requirement constantly expanded;
(2) the present invention uses unique fermented bacterium and culture process, and only need to add a small amount of inorganic ion at fermenting process, cost is low, gluconic acid concentration and productive rate high, be convenient to later separation and purifying.
(3) the present invention is that raw material high-efficiency fermenting produces gluconic acid with lignocellulose, existing cellulose ethanol production correlation technique can be used for reference, realize producing gluconic acid and salt compounds thereof from lignocellulose, realize the large fine chemicals biorefinery based on lignocellulose.
[accompanying drawing explanation]
Fig. 1 embodiment 1 fermenting process curve;
Fig. 2 embodiment 2 fermenting process curve
[embodiment]
A kind of bio-transformation lignocellulose of the present invention is below provided to produce the embodiment of the method for gluconic acid.
Embodiment 1
By the dilution heat of sulfuric acid of maize straw and 5% (w/w) concentration of drying with 2: 1 mass ratio mix, in 190 DEG C, pre-treatment 3min under the condition of 1.2MPa; After pre-treatment, material pH value is adjusted to 5.5, accesses cultured biological detoxication bacterial classification Amorphotheca resinae ZN1 (CGMCC7452), quiescent culture 7d under the condition of 25 DEG C; Then the material after detoxification is added in reactor, regulate solids content to 25% (w/w), add cellulase (consumption is 15FPU/g DM), in 50 DEG C, saccharification 48h under the condition of 150rpm, in saccharifying with 5M sodium hydroxide control pH 4.8.In the corn stalk hydrolysis obtained, the concentration of glucose is 71.42g/L.Corn stalk hydrolysis after the centrifugal 10min of 10000rpm as substratum, with 10% inoculum size access aspergillus niger (Aspergillus niger M276, purchased from Shanghai City Industry Wei Biological Research Institute DSMZ) seed culture fluid, 30 DEG C of liquid state fermentation 96h, rotating speed 200rpm, in fermented liquid, gluconic acid concentration is 70.6g/L, and gluconic acid yield reaches 98%.
Embodiment 2
By the dilution heat of sulfuric acid of maize straw and 5% (w/w) concentration of drying with 2: 1 mass ratio mix, in 190 DEG C, pre-treatment 3min under the condition of 1.2MPa; Pretreated material is directly joined in reactor, regulate solids content to 25% (w/w), add cellulase (consumption is 15FPU/gDM), in 50 DEG C, saccharification 48h under the condition of 150rpm, in saccharifying with 5M sodium hydroxide control pH 4.8.In the corn stalk hydrolysis obtained, the concentration of glucose is 75.36g/L.Corn stalk hydrolysis after the centrifugal 10min of 10000rpm as substratum, with 10% inoculum size access aspergillus niger (Aspergillus niger M276, purchased from Shanghai City Industry Wei Biological Research Institute DSMZ) seed culture fluid, 30 DEG C of liquid state fermentation 168h, rotating speed 200rpm, in fermented liquid, gluconic acid concentration is 79.04g/L, is 95% to the transformation efficiency of glucose.
Cellulose raw material involved in the present invention is not limited to maize straw, can also be the one in wheat straw, straw, wood chip, energy-source plant or forestry waste; Lignocellulose handled thing is selected from the one adopted in swollen quick-fried, the liquefied ammonia of diluted acid, steam swollen quick-fried, the pretreated wheat straw of biological process, straw, wood chip, energy-source plant, forestry waste.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, without departing from the inventive concept of the premise; can also make some improvements and modifications, these improvements and modifications also should be considered within the scope of protection of the present invention.
Claims (8)
1. the method for a bio-transformation lignocellulose production gluconic acid, it is characterized in that, be that raw material low-cost high-efficiency produces gluconic acid with lignocellulose, specifically first carry out the pre-treatment of lignocellulosic material, the raw material obtained carries out detoxification or not detoxification, then produce gluconic acid through following two kinds of approach: one, obtain saccharified liquid with the raw material after cellulase degradation process, then access gluconic acid production bacterial strain and carry out gluconic acid fermentation; Two, raw material, cellulase and the gluconic acid after process is produced bacterial strain to add in reactor simultaneously, produce gluconic acid by the mode of synchronous saccharification and fermentation.
2. a kind of bio-transformation lignocellulose as claimed in claim 1 produces the method for gluconic acid, and it is characterized in that, its concrete steps are:
(1) lignocellulose pre-treatment:
Lignocellulose pre-treatment comprises dilute acid pretreatment, steam explosion, and swollen quick-fried, the sulphite pre-treatment of the explosion of ammonia fiber, liquefied ammonia, alkaline process pre-treatment, machinery grind pre-treatment, Biological Pretreatment; Preferred dilute acid pretreatment method, dilute acid pretreatment refers to that dilution heat of sulfuric acid infiltrates material, then at 180 ~ 200 DEG C, maintains 1 ~ 10min, destroys lignocellulose structure, thus is conducive to follow-up enzymolysis process;
(2) liquid nutrient medium is utilized to cultivate aspergillus niger:
40 ~ 80g/L glucose, 0 ~ 1g/L magnesium sulfate, 0 ~ 2g/L SODIUM PHOSPHATE, MONOBASIC, the potassium primary phosphate of 0 ~ 2g/L, the manganous sulfate of 0 ~ 2g/L, liquid culture aspergillus niger 25 ~ 40 DEG C, 50 ~ 500rpm, cultivation is 12 ~ 48h, and cell concentration reaches 1 ~ 3g/L.The aspergillus niger cultivated is used for further cultivation or the fermentation of liquid or solid fermented substrate;
(3) solid-state biological detoxication:
After regulating lignocellulosic material pH and water content, access detoxification mould Amorphotheca resinaeZN1, deposit number CGMCC7452, solid state rheology is characterised in that culture temperature 20 ~ 35 DEG C, and material moisture is 30 ~ 70%, material pH4.5 ~ 6.5.The compound that detoxification removes comprises formic acid, acetic acid, furfural, hydroxymethylfurfural etc.;
(4) production of gluconic acid:
Add lignocellulosic material and the cellulase of detoxification or non-detoxification, water use regulation solids content, at 50 DEG C in fermentor tank simultaneously, saccharification 48 ~ 96h under the condition of pH4.8, solid-liquid separation is carried out to the slurries after saccharification, obtains supernatant liquor, i.e. lignocellulose enzymolysis liquid; The aspergillus niger strain that liquid culture is good, be inoculated in hydrolyzed solution and ferment;
Or the aspergillus niger strain, lignocellulosic material (detoxification or non-detoxification) and the cellulase that liquid culture are obtained join in reactor simultaneously, at 30 ~ 50 DEG C, synchronous saccharification and fermenting lignocellulose raw material production gluconic acid under the condition of pH4 ~ 6, fermentation time is 72 ~ 120h.
3. a kind of bio-transformation lignocellulose as claimed in claim 2 produces the method for gluconic acid, it is characterized in that, in described step (1), described lignocellulosic material comprises one or more the mixing such as maize straw, wheat-straw, straw, Cotton Stalk, energy-source plant, wood, wood chip, waste paper, agroforestry waste.
4. a kind of bio-transformation lignocellulose as claimed in claim 2 produces the method for gluconic acid, it is characterized in that, in described step (3), if bacterium produced by the gluconic acid selecting inhibition tolerance strong, then step (3), namely can omit the detoxification treatment of lignocellulosic material.
5. a kind of bio-transformation lignocellulose as claimed in claim 2 produces the method for gluconic acid, it is characterized in that, in described step (3), if bacterium produced by the gluconic acid selecting inhibition tolerance poor, then pretreated lignocellulosic material need through detoxification treatment, remove the inhibition produced in preprocessing process, comprise organic acid compounds, poison-removing method includes but not limited to biological detoxication, washing, crosses alkaline purification (Overliming), charcoal absorption, ionic adsorption, preferred biological detoxication.
6. a kind of bio-transformation lignocellulose as claimed in claim 2 produces the method for gluconic acid, it is characterized in that, in described step (4), the massfraction of described lignocellulose in fermentor tank is 10 ~ 40%, cellulase is 5 ~ 30FPU/g DM, leavening temperature 30 ~ 50 DEG C.
7. a kind of bio-transformation lignocellulose as claimed in claim 2 produces the method for gluconic acid, it is characterized in that, in described step (4), the bacterial strain producing grape acid mainly contains aspergillus niger, mould, glucose oxidation and bacillus etc., and wherein aspergillus niger is more suitable for utilizing lignocellulosic material to produce gluconic acid.
8. a kind of bio-transformation lignocellulose as claimed in claim 2 produces the method for gluconic acid, it is characterized in that, in described step (4), the gluconic acid that fermentation generates, comprise the gluconic acid of free state, and the gluconate regulating fermentation pH to generate.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106906265A (en) * | 2015-12-22 | 2017-06-30 | 天津科技大学 | A kind of straw pretreatment method produced without waste liquid |
| CN106957875A (en) * | 2016-01-11 | 2017-07-18 | 中国石油化工股份有限公司 | A kind of method of utilization lignocellulosic material fermenting and producing butanol |
| CN107557397A (en) * | 2017-08-24 | 2018-01-09 | 南京林业大学 | A kind of method of hydrogen peroxide regulation and control whole-cell catalytic glucose controlled syntheses 2- ketone group gluconic acids |
| CN109554409A (en) * | 2017-09-25 | 2019-04-02 | 华东理工大学 | A kind of solid-liquid separating method of lignocellulosic material simultaneous saccharification and fermentation wine with dregs |
| CN109971806A (en) * | 2017-12-28 | 2019-07-05 | 南京理工大学 | Mix lignocellulose pretreatment method and its zymotechnique |
| CN110564781A (en) * | 2019-10-23 | 2019-12-13 | 东北农业大学 | Pretreatment method of straw lignocellulose raw material |
| CN112941117A (en) * | 2020-12-23 | 2021-06-11 | 华东理工大学 | Method for synthesizing L-lactide from chiral L-lactic acid produced by using lignocellulose biomass as raw material |
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| CN106906265A (en) * | 2015-12-22 | 2017-06-30 | 天津科技大学 | A kind of straw pretreatment method produced without waste liquid |
| CN106957875A (en) * | 2016-01-11 | 2017-07-18 | 中国石油化工股份有限公司 | A kind of method of utilization lignocellulosic material fermenting and producing butanol |
| CN106957875B (en) * | 2016-01-11 | 2020-04-10 | 中国石油化工股份有限公司 | Method for producing butanol by fermenting lignocellulose raw material |
| CN107557397A (en) * | 2017-08-24 | 2018-01-09 | 南京林业大学 | A kind of method of hydrogen peroxide regulation and control whole-cell catalytic glucose controlled syntheses 2- ketone group gluconic acids |
| CN109554409A (en) * | 2017-09-25 | 2019-04-02 | 华东理工大学 | A kind of solid-liquid separating method of lignocellulosic material simultaneous saccharification and fermentation wine with dregs |
| CN109971806A (en) * | 2017-12-28 | 2019-07-05 | 南京理工大学 | Mix lignocellulose pretreatment method and its zymotechnique |
| CN110564781A (en) * | 2019-10-23 | 2019-12-13 | 东北农业大学 | Pretreatment method of straw lignocellulose raw material |
| CN112941117A (en) * | 2020-12-23 | 2021-06-11 | 华东理工大学 | Method for synthesizing L-lactide from chiral L-lactic acid produced by using lignocellulose biomass as raw material |
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| CN104805137B (en) | 2018-11-23 |
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