CN104805137B - A kind of method of bioconversion lignocellulosic production gluconic acid - Google Patents
A kind of method of bioconversion lignocellulosic production gluconic acid Download PDFInfo
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- CN104805137B CN104805137B CN201410034928.1A CN201410034928A CN104805137B CN 104805137 B CN104805137 B CN 104805137B CN 201410034928 A CN201410034928 A CN 201410034928A CN 104805137 B CN104805137 B CN 104805137B
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- 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
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/58—Aldonic, ketoaldonic or saccharic acids
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- C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
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Abstract
The present invention relates to a kind of methods that inventor utilizes aspergillus niger bioconversion lignocellulosic efficiently to produce gluconic acid.The method of offer is specifically related to, and cultured aspergillus niger is inoculated in fermenting and producing gluconic acid in ligno-cellulose hydrolysate, gluconic acid concentration is up to 80g/L after fermentation 96 hours；Or the good aspergillus niger of Liquid Culture, lignocellulosic material and cellulase are added in reactor simultaneously, synchronize saccharification and fermenting and producing gluconic acid.The present invention has the characteristics that the time is short, high-efficient, and lignocellulosic material is from a wealth of sources and cheap, has very bright development prospect and application prospect.
Description
【Technical field】
Biology manufacture and biorefinery field the present invention relates to biological-based chemicals, specifically, being corn stover etc.
Method of the lignocellulosic material by saccharification and fermenting and producing high concentration glucose acid.
【Background technique】
Gluconic acid and its esters have the characteristics that nontoxic, thermal stability is good and is not easy to deliquesce, chemical industry, food, medicine,
The industries such as light industry have been widely used.Especially in terms of cement additive, sodium gluconate can be used as water-reducing agent and slow setting
Agent and be widely used, and achieve good effect；It can be used as nutritional supplement, curing agent, buffer etc. in food industry,
Present people have developed the condiment of high flavor using sodium gluconate as raw material.Further, since gluconic acid has
It is more that excellent chelating ability is also widely used for water treatment, plating, metal and nonmetallic surface clean and manufacture of cement etc.
Kind industrial department.Currently, the annual requirement of gluconic acid alreadys exceed 500,000 tons.
The method of production gluconic acid mainly has biological fermentation process, eletrooxidation method, electrolytic oxidation and multiphase
Four kinds of methods of catalytic oxidation.The intermediate steps of eletrooxidation method are more, and by-product is more, and product is difficult to separate, and finished product
In contain sodium chloride, make its in terms of cement additive application be restricted；Electrolytic oxidation energy consumption is high in production and
It is not easy to control；Heterogeneous catalytic oxidation method needs to cause using expensive catalyst the production cost of gluconic acid high.And
Microbe fermentation method is since fermenting speed is fast, fermentation process is easily controllable, product easily extracts, produces and stablizes mild, easy industry
The advantages that metaplasia produces, and become the most important production method of glucose present acid.
The raw material of fermenting and producing gluconic acid is mainly glucose, molasses and starchy material.And current gluconic acid
Yield is not able to satisfy the increasingly increased demand to gluconic acid, and the enlarging extensive simply of starch deep processing enterprise will certainly
Seriously affect the grain security of country.Therefore, seeking cheap and from a wealth of sources gluconic acid raw materials for production is grape
The matter of utmost importance that saccharic acid industry healthy development is faced.China is large agricultural country, the lignocellulosic materials such as agriculture and forestry waste
Yield it is huge, but it is most of all slatterned by inefficient burning, not only cause the huge wasting of resources, but also result in ratio
More serious environment and traffic problems.And lignocellulosic material is utilized to produce gluconic acid, then it can effectively solve glucose
The raw material sources problem of sour industry, while current rural economy, rural development and rural demography can be effectively relieved, it is provided for the deep processing of agricultural wastes
New outlet.
Bioconversion lignocellulosic material production of chemicals, generally comprises pretreatment of raw material, enzyme hydrolysis, fermentation and product
The processes such as separation.Lignocellulosic material destroys its compact texture by pretreatment, promotes cellulase to the drop of cellulose
Solution.But preprocessing process generates the mortifiers such as organic acid, aldehydes and phenols.In order to improve fermenting for cellulose matrix
Property, mortifier therein must be reduced or removed completely before fermentation.But grape is produced by substrate fermentation of lignocellulosic
There are many technological difficulties for saccharic acid, at present the method in relation to using in the report using lignocellulosic material production gluconic acid
Mostly heterogeneous catalytic oxidation method.During producing gluconic acid using these methods, need to use a large amount of sulfuric acid, hydroxide
The chemicals such as sodium, and catalyst is mostly Na2Cu(OH)4With gold/phosphorus heteropoly tungstic acid cesium salt, although this method high conversion rate,
Used catalyst is limited in cycle-index, and the catalyst consumption of unit product is high, in a disguised form increases the life of gluconic acid product
Produce cost.
The present invention provides the pretreatment using complete true lignocellulosic material as matrix, proposed using inventor,
Saccharification and zymotechnique production high concentration glucose acid.
【Summary of the invention】
Present invention aims at the fermentation substrate for expanding crude dextrose acid fermentation, use is cheap, resourceful
Lignocellulosic, provide one kind using lignocellulosic as raw material, pass through saccharification and the side of fermenting and producing high concentration glucose acid
Method.
Design of the invention：Gluconic acid is produced by raw material low-cost high-efficiency of lignocellulosic, specifically first
The pretreatment for carrying out lignocellulosic material, obtained raw material carry out detoxification or not detoxification, then pass through following two approach
Produce gluconic acid：One, with cellulase degradation, treated that raw material obtains saccharified liquid, then accesses gluconic acid and produces bacterial strain
Carry out gluconic acid fermentation；Two, by treated, reactor is added in raw material, cellulase and gluconic acid production bacterial strain simultaneously
In, gluconic acid is produced by way of synchronous saccharification and fermentation.
The purpose of the present invention is achieved through the following technical solutions：
A kind of method of bioconversion lignocellulosic production gluconic acid, which is characterized in that the specific steps are that：
(1) lignocellulosic pre-processes：
Lignocellulosic pretreatment includes dilute acid pretreatment, steam blasting, the explosion of ammonia fiber, the swollen quick-fried, sulphite of liquefied ammonia
Pretreatment, alkaline process pretreatment, machinery grind pretreatment, Biological Pretreatment；It is preferred that dilute acid pretreatment method, dilute acid pretreatment refer to
Dilution heat of sulfuric acid infiltrates material, and 1~10min is then maintained at 180~200 DEG C, lignocellulosic structure is destroyed, thus favorably
In subsequent enzymolysis process.
The lignocellulosic material includes corn stover, wheat-straw, straw, Cotton Stalk, energy-source plant, wood, wood
The mixing of the one or more such as bits, waste paper, agriculture and forestry waste.
The lignocellulosic material have passed through pretreatment, be destroyed its compact texture, so that improving it can digest
Property.Preprocess method includes but is not limited to that dilute acid pretreatment, steam blasting, the explosion of ammonia fiber, the swollen quick-fried, sulphite of liquefied ammonia are located in advance
Reason, alkaline process pretreatment, machinery grind pretreatment, Biological Pretreatment etc., preferably dilute acid pretreatment method.
(2) fluid nutrient medium culture aspergillus niger is utilized：
40~80g/L glucose, 0~1g/L magnesium sulfate, 0~2g/L sodium dihydrogen phosphate, the potassium dihydrogen phosphate of 0~2g/L, 0
The manganese sulfate of~2g/L, is cultivated as 12~48h by 25~40 DEG C of Liquid Culture aspergillus niger, 50~500rpm, and cell concentration reaches 1
~3g/L.Further culture or fermentation of the aspergillus niger of culture for liquid or solid fermentation substrate.
(3) solid-state biological detoxication：
After adjusting lignocellulosic material pH and water content, detoxification mould Amorphotheca resinae ZN1 is accessed,
Deposit number CGMCC7452, solid state rheology are characterized in that 20~35 DEG C of cultivation temperature, and material moisture is 30~70%, material
PH4.5~6.5.The compound that detoxification is removed includes formic acid, acetic acid, furfural, hydroxymethylfurfural etc..
If the gluconic acid for selecting mortifier tolerance strong produces bacterium, step (3), i.e., to lignocellulosic material
Detoxification treatment can be omitted.
If the gluconic acid for selecting mortifier tolerance poor produces bacterium, pretreated lignocellulosic material is needed
By detoxification treatment, the mortifier generated in preprocessing process, including organic acid compounds are removed, such as formic acid, acetic acid, acetyl
Propionic acid etc., furan derivative, such as furfural, hydroxymethylfurfural, phenolic compound, such as parahydroxyben-zaldehyde, vanillic aldehyde.It is de-
Malicious method includes but is not limited to biological detoxication, washing, crosses alkali process (Overliming), activated carbon adsorption, ionic adsorption etc., excellent
Select biological detoxication.
(4) production of gluconic acid：
The lignocellulosic material and cellulase of detoxification or non-detoxification is added simultaneously into fermentor, water use regulation is solid
Body content, at 50 DEG C, be saccharified 48~96h under conditions of pH4.8, is separated by solid-liquid separation to the slurries after saccharification, obtains supernatant,
I.e. lignocellulosic digests liquid；The good aspergillus niger strain of Liquid Culture, it is inoculated in hydrolyzate and ferments.
Aspergillus niger strain, lignocellulosic material (detoxification or non-detoxification) and the cellulose that either Liquid Culture is obtained
Enzyme is added in reactor simultaneously, and at 30~50 DEG C, synchronous saccharification and fermenting lignocellulose raw material are raw under conditions of pH4~6
Malaga saccharic acid, fermentation time are 72~120h.
The mass fraction of the lignocellulosic in the fermenter is 10~40%, and cellulase is 5~30FPU/g
DM, 30~50 DEG C of fermentation temperature.The cellulose raw material is selected from solid straws raw material or solid straws processed material；Wherein：
The cellulose raw material includes in corn stover, wheat straw, straw, sawdust, energy-source plant or forestry waste.
Production grape acid bacterial strain mainly have aspergillus niger, mould, glucose oxidation and bacillus etc., wherein aspergillus niger more suitable for
Gluconic acid is produced using lignocellulosic material.
Ferment the gluconic acid generated, the gluconic acid including free state, and adjusts fermentation pH glucose generated
Hydrochlorate, such as sodium gluconate, calcium gluconate, ammonium gluconate etc..
Compared with prior art, the positive effect of the present invention is：
(1) for the present invention using lignocellulosic as fermenting raw materials gluconic acid, matrix is low in cost, from a wealth of sources, sufficiently benefit
With renewable resource, grain is not striven with the mankind, can satisfy the ever-expanding market demand；
(2) present invention uses unique fermenting microbe and culture process, only needs to add a small amount of inorganic salts in fermentation process
Ion, at low cost, gluconic acid concentration and yield are high, are convenient for later separation and purifying.
(3) present invention produces gluconic acid by raw material high-efficiency fermenting of lignocellulosic, can use for reference existing cellulose ethanol
The relevant technologies are produced, realizes from lignocellulosic and produces gluconic acid and its salt compounds, are realized based on lignocellulosic
Large fine chemicals biorefinery.
【Detailed description of the invention】
1 fermentation process curve of Fig. 1 embodiment；
2 fermentation process curve of Fig. 2 embodiment
【Specific embodiment】
A kind of specific embodiment of the method for bioconversion lignocellulosic production gluconic acid of the present invention presented below.
Embodiment 1
The corn stover of drying is uniformly mixed with the dilution heat of sulfuric acid of 5% (w/w) concentration with 2: 1 mass ratio, in 190
DEG C, pre-process 3min under conditions of 1.2MPa；Material pH value is adjusted to 5.5 after pretreatment, accesses cultured biological detoxication
Strain Amorphotheca resinae ZN1 (CGMCC7452), the stationary culture 7d under conditions of 25 DEG C；It then will be after detoxification
Material be added in reactor, adjust solid content to 25% (w/w), cellulase (dosage is 15FPU/g DM) be added, in
50 DEG C, be saccharified under conditions of 150rpm 48h, with 5M sodium hydroxide control pH 4.8 in saccharifying.Obtained corn stover water
The concentration for solving glucose in liquid is 71.42g/L.Corn stalk hydrolysis is used as culture medium after 10000rpm is centrifuged 10min,
With 10% inoculum concentration access aspergillus niger, (Aspergillus niger M276 is purchased from Shanghai City Industry Wei Biological Research Institute strain
Collection) seed culture fluid, 30 DEG C of liquid state fermentation 96h, revolving speed 200rpm, gluconic acid concentration is 70.6g/L in fermentation liquid,
Gluconic acid yield reaches 98%.
Embodiment 2
The corn stover of drying is uniformly mixed with the dilution heat of sulfuric acid of 5% (w/w) concentration with 2: 1 mass ratio, in 190
DEG C, pre-process 3min under conditions of 1.2MPa；Pretreated material is added directly into reactor, adjust solid content to
25% (w/w) is added cellulase (dosage 15FPU/gDM), and be saccharified 48h under conditions of 50 DEG C, 150rpm, saccharifying
It is middle to control pH 4.8 with 5M sodium hydroxide.The concentration of glucose is 75.36g/L in obtained corn stalk hydrolysis.Corn stalk
Stalk hydrolyzate is used as culture medium after 10000rpm is centrifuged 10min, accesses aspergillus niger (Aspergillus with 10% inoculum concentration
Niger M276 is purchased from Shanghai City Industry Wei Biological Research Institute Culture Collection Center) seed culture fluid, 30 DEG C of liquid state fermentations
168h, revolving speed 200rpm, gluconic acid concentration is 79.04g/L in fermentation liquid, and the conversion ratio to glucose is 95%.
Cellulose raw material according to the present invention is not limited to corn stover, can also plant for wheat straw, straw, sawdust, the energy
One of object or forestry waste；Lignocellulosic processed material be selected from using diluted acid, steam it is swollen it is quick-fried, liquefied ammonia is swollen quick-fried, biological
One of the pretreated wheat straw of method, straw, sawdust, energy-source plant, forestry waste.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, without departing from the inventive concept of the premise, can also make several improvements and modifications, these improvements and modifications also should be regarded as
In protection scope of the present invention.
Claims (2)
1. a kind of method of bioconversion lignocellulosic production gluconic acid, which is characterized in that include the following steps：
By the dilution heat of sulfuric acid of the corn stover of drying and 5%w/w concentration with 2:1 mass ratio is uniformly mixed, in 190 DEG C,
3min is pre-processed under conditions of 1.2MPa；Material pH value is adjusted to 5.5 after pretreatment, accesses cultured biological detoxication strain
Amorphotheca resinae ZN1CGMCC7452, the stationary culture 7d under conditions of 25 DEG C；Then by the material after detoxification
Be added in reactor, adjust solid content to 25%w/w, cellulase is added, dosage is 15FPU/g DM, in 50 DEG C,
Be saccharified 48h under conditions of 150rpm, with 5M sodium hydroxide control pH 4.8 in saccharifying；Obtained corn stalk hydrolysis
The concentration of middle glucose is 71.42g/L；Corn stalk hydrolysis is used as culture medium after 10000rpm is centrifuged 10min, with
10% inoculum concentration accesses aspergillus niger；Seed culture fluid, 30 DEG C of liquid state fermentation 96h, revolving speed 200rpm, gluconic acid is dense in fermentation liquid
Degree is 70.6g/L, and gluconic acid yield reaches 98%；
The aspergillus niger Aspergillus niger M276 is purchased from Shanghai City Industry Wei Biological Research Institute Culture Collection Center.
2. a kind of method of bioconversion lignocellulosic production gluconic acid, which is characterized in that include the following steps：
By the dilution heat of sulfuric acid of the corn stover of drying and 5%w/w concentration with 2:1 mass ratio is uniformly mixed, in 190 DEG C,
3min is pre-processed under conditions of 1.2MPa；Pretreated material is added directly into reactor, adjust solid content to
Cellulase is added in 25%w/w, and dosage is 15FPU/g DM, and be saccharified 48h under conditions of 50 DEG C, 150rpm, in saccharifying
With 5M sodium hydroxide control pH 4.8；The concentration of glucose is 75.36g/L in obtained corn stalk hydrolysis；Corn stover
Hydrolyzate by 10000rpm be centrifuged 10min after be used as culture medium, with 10% inoculum concentration access aspergillus niger seed culture fluid, 30 DEG C
Liquid state fermentation 168h, revolving speed 200rpm, gluconic acid concentration is 79.04g/L in fermentation liquid, and the conversion ratio to glucose is
95%；
The aspergillus niger Aspergillus niger M276 is purchased from Shanghai City Industry Wei Biological Research Institute Culture Collection Center.
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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 |
| 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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