CN102286571B - Clean and high-efficiency production process for preparing xylose and L-arabinose - Google Patents

Abstract

The invention relates to a clean and high-efficiency production process for preparing xylose and L-arabinose and belongs to the field of application of wastes of agriculture and forestry. The production process comprises the following steps of: performing pretreatment on raw materials; hydrolyzing the raw materials in a high-temperature cooking mode; performing neutralizing treatment and performing centrifugal separation on solid slag to obtain xylose or xylooligosaccharide hydrolyzate; adding coarse enzyme solution into xylooligosaccharide solution to hydrolyze to obtain xylose solution; treating the hydrolyzate by Angle yeast; purifying by using active carbon and resin; performing recompression and concentration treatment on the purified hydrolyzate by using a mechanical vapor recompression (MVR) machine; crystallizing the concentrated solution in a gradient program cooling mode to obtain xylose products and xylose mother solution; separating the xylose mother solution by a simulated moving bed or a chromatographic separation technology to obtain a xylose phase and an L-arabinose phase; returning the xylose phase to the xylose solution to concentrate and recrystallize; reusing the xylose mother solution; performing MOVR concentration treatment on the L-arabinose phase and crystallizing in a gradient program cooling mode to obtain L-arabinose products; and reusing the L-arabinose mother solution.

Description

A kind of production technique of preparing wood sugar and L-arabinose of clean and effective

Technical field

The present invention relates to a kind of technique of utilizing the raw material cleaner production wood sugars such as agriculture and forestry organic waste material and L-arabinose, especially raw material is taked after pre-treatment high temperature steaming hydrolysis process to process the agriculture and forestry organic waste materials such as cotton seed hulls and obtained xylose hydrolysis fluid through microbial method removal of impurities sugar and purification process post crystallization acquisition wood sugar crystal, xylose mother liquid obtains the method for wood sugar and L-arabinose crystal through simulation moving-bed separated post crystallization.

Background technology

The product that carries out photosynthesis every year on the earth and obtain is up to nearly 2,000 hundred million tons, is unique utilizability resource that can ultra-large regeneration on the earth.Accounting in phytomass most lignocelluloses is to enrich the most in the world, and the most cheap renewable resources, wherein by less than 12% that people utilized.China Shi Yige large agricultural country, the agriculture and forestry organic waste material raw material that is rich in lignocellulose that can be used for producing wood sugar is very abundant.It's a pity that more these raw materials are dropped or burn, not only cause the huge wasting of resources, also caused serious environmental pollution.Therefore carry out the research of the production technique of the rare sugar alcohols such as wood sugar and L-arabinose, to improving china natural resources and energy utilization rate, help farmer richness extra earning to there is important and positive meaning, this research invention not only has higher direct economic benefit, and has considerable social benefit.

Wood sugar, as a kind of base-material of functional nutrient, has almost zero energy value, the bifidus bacillus in activating human body enteron aisle short its growth, and the immunological competence of raising body, preventing dental caries, reducing blood-fat, reduces the effect of cholesterol and prevention intestinal cancer.The compatibleness of wood sugar and food is fine, is also having certain purposes aspect light industry, chemical industry.

L-arabinose is a kind of efficient functional nutrient additive, if add 3.5% content in sucrose, can suppress the absorption of 40% sucrose, makes blood glucose value raise less approximately 50%.Also can suppress fat, preventing constipation, promotes bifidobacterium growth, and has the effect etc. that changes skeletal muscle fibre composition.Meanwhile, L-arabinose is a kind of important medicine intermediate, is mainly used to medicine synthetic anticancer, antiviral and treatment cardiovascular and cerebrovascular diseases, can also be as biochemical reagents, for the preparation of bacteria culture medium, or carry out the synthetic of spices, be widely used in food, healthcare products and field of medicaments.

In the process of hydrolysis of hemicellulose, pretreatment technology is an important link, is also one of the effect of restriction hydrolysis and important factor of economy.In order to improve hemicellulose saccharification efficiency, reduce its cost, to agriculture and forestry organic waste material, raw-material pre-treatment becomes most important.In whole pretreatment technology, peeling off or degrading of xylogen is vital.The remaining rate of xylogen is very large to agriculture and forestry organic waste material pre-treatment cost impact, whole except delignification very large with its cost difference of xylogen of removing 20-65%, much research shows, as long as according to different material characteristics, the xylogen of removing 20-65% just can improve hemicellulose hydrolysis efficiency and saccharification productive rate significantly.Preconditioning technique must meet following condition simultaneously.1. improve hydrolysis productive rate and the efficiency of hemicellulose; 2. avoid producing the generation to environment or the inhibited by product of subsequent technique; 3. avoid degraded and the loss of carbohydrate; 4. economic feasibility is strong, uses as much as possible the technique means of less energy-consumption, to reduce process costs.At present, preconditioning technique mainly comprises Physical, chemical method, biological process, and some integrated conduct methods such as physical-chemical method.

Industrial traditional hydrolysis of hemicellulose prepare wood sugar process using be the method for sulfuric acid liquid hydrolysis material, whole production process produces and in a large amount of saline sewages and N-process, produces a large amount of waste residues, 1 ton of wood sugar saline sewage quantity discharged of every production is up to more than 300 tons, and pollutent is difficult to biochemical treatment, environmental pollution is serious.The present invention adopt simple and practical pretreatment technology in conjunction with the high temperature steaming of clean and effective or boiling in conjunction with the method for enzymolysis, the hemicellulose in agriculture and forestry organic waste material raw material is hydrolyzed and produces the technique of xylose hydrolysis fluid, substitute classical acid hydrolysis process.Whole production technique produces without neutralization and without waste residue, and the quantity discharged of the sewage in hydrolytic process has obtained significantly reducing, and 1 ton of wood sugar quantity of wastewater effluent of every production is less than 50 tons.The xylose hydrolysis fluid making with traditional sulfuric acid process is compared saltiness and has been reduced by 80%.

Agriculture and forestry organic waste material is in the process of hydrolysis, the main component of hemicellulose hydrolysate is wood sugar, has in addition other assorted sugar such as glucose, L-arabinose, and the liquid glucose viscosity not only obtaining after concentrated is very large, and the not high crystallization yield of xylose that causes of the content of wood sugar is lower, purity is affected.The xylose mother liquid obtaining after crystallization is because assorted sugared content is at raise to some extent through ratio after crystallization (especially glucose), xylose crystallization mother liquor cannot continue to reply utilization, L-arabinose cannot obtain effective purifying, makes the effective constituent waste in raw material serious.Therefore how improving productive rate and the crystal purity of xylose hydrolysis fluid crystalline xylose, make full use of the effective constituent in xylose mother liquid simultaneously, reduce production costs, is a problem demanding prompt solution.There are in recent years some to utilize yeast fermentation to process the glucose in xylose mother liquid, recycle simulation moving-bed by the wood sugar report that obtain high added value monose separated with L-arabinose, but because assorted sugared concentration in mother liquor is too high, osmotic pressure is too large, impact for thalli growth and removal glucose is larger, cause also needing mother liquor Macrodilution in fermenting process, increased the weight of the load of processing.

Summary of the invention

The present invention is directed to above problem, before being chosen in xylose crystalline process, utilize yeast saccharomyces cerevisiae to process the assorted sugar such as glucose in xylose hydrolysis fluid.Because the sugared concentration in hydrolyzed solution is now lower, osmotic pressure is suitable, and the assorted sugar such as glucose that thalline can fully consume are wherein grown, thereby efficiently solves assorted sugar for the impact of xylose crystalline, has increased substantially yield and the purity of crystallization.Meanwhile, in the xylose mother liquid obtaining after crystallization, only contain other carbohydrates of wood sugar, L-arabinose and minute quantity, can directly utilize some separation means to obtain L-arabinose.This technology has improved the productive rate of xylose product effectively, has optimized the treatment process of xylose mother liquid, has reduced production cost, makes raw material obtain more adequately and reasonably utilizing.At present in sugar alcohol industrial production, relevant for xylose mother liquid crystallization, produced the technology report of L-arabinose, but mostly exist product crystallization yields low, the problem such as the not high and rate of recovery of product purity is low, the present invention adopts the technology of simulation moving-bed separation chromatography separation, to the wood sugar in xylose mother liquid, carry out separated with L-arabinose mixed system, obtain respectively the independent two-phase liquid of wood sugar and L-arabinose, whole separating technology level of automation is high, simple to operate, the utilization ratio of separating filler is high, the resolution of wood sugar and L-arabinose can reach more than 95%, L-arabinose after separation carries out condensing crystal mutually again, the yield of product and purity have had significantly lifting.

A production technique of preparing wood sugar and L-arabinose for clean and effective, is characterized in that, comprises the steps (seeing Figure of description 1):

1. a production technique of preparing wood sugar and L-arabinose for clean and effective, is characterized in that, comprises the steps:

(1) agriculture and forestry organic waste material raw material carries out pre-treatment and dipping pretreatment; Described agriculture and forestry organic waste material raw material comprises: corn cob, maize straw, rice straw, jowar stalk, cotton seed hulls or bagasse;

Pre-treating technology comprises: raw-material pulverizing, dedusting, washing three process;

The soak solution that described dipping pretreatment starting material are used is dilute sulphuric acid, dilute hydrochloric acid or rare oxalic acid, the massfraction of dilute sulphuric acid, dilute hydrochloric acid or rare oxalic acid is 0.1-0.5%, soak solution is 4-8 with the massfraction ratio of raw material: 1, the temperature of soaking is 30-100 ℃, and the time of immersion is 6-24h;

After dipping pretreatment finishes, solid-liquid separation obtains pretreated raw material, and the mode of isolated pretreated raw material by washing regulates pH value to 4.5-6.0, and soak solution is replied and utilized;

(2) the agriculture and forestry organic waste material raw material after process (1) takes the method hydrolyzed hemicellulose of high temperature steaming to obtain high temperature steaming Xylose or high temperature steaming xylo-oligosaccharide liquid, specific as follows;

Pretreated agriculture and forestry organic waste material is added in reaction unit, adds 5-20 doubly to the water of solid material dry weight; Initial pH is 5.0-6.0; Boiling 0.5-2h under the condition of 145 ℃-155 ℃; Mixing speed is 80-150rpm; Operating pressure is 0.5-1.0MPa, obtains xylo-oligosaccharide cooking liquor;

Or the agriculture and forestry organic waste material raw materials such as pretreated corn cob or cotton seed hulls are added in reaction unit, add 5-20 doubly to the water of solid material dry weight; At initial pH, be 4.5-6.0; High temperature steaming 1-3h under the condition of 160-180 ℃; Mixing speed is 80-150rpm; Operating pressure is 0.6-1.2MPa, obtains wood sugar cooking liquor;

(3) utilize milk of lime to above-mentioned high temperature steaming xylo-oligosaccharide liquid carried out neutralizing treatment to pH at 8.0-9.0, again with phosphoric acid or hydrochloric acid readjustment pH to 6.0, Plate Filtration obtains supernatant liquor afterwards;

Xylo-oligosaccharide cooking liquor after the crude enzyme liquid that employing aspergillus niger produces is processed (3) carries out enzymolysis and obtains wood sugar enzymolysis solution, specific as follows:

Cha Shi substratum, 115 ℃ of sterilizing 30min, access aspergillus niger is cultivated the aspergillus niger seed liquor that obtains having spore shape form for 3-4 days under the condition of 28-30 ℃; Product enzymic fermentation is cultivated: sucrose 2.4g, MgSO ₄0.12g, K ₂hPO ₄0.12g, KH ₂pO ₄0.24g, FeSO ₄0.0012g, NaNO ₃0.012g, uses 100ml water dissolution, mixes with the mixture of wheat bran with corn cob, and the mixture of corn cob and wheat bran amounts to 100g, and corn cob and wheat bran mass ratio are 3: 7-7: 3, and under the condition of 115 ℃, sterilizing 30min obtains fermention medium;

Under the condition of aseptic technique, in 100g fermention medium, add the aspergillus niger seed liquor 10-15ml with spore shape form, add sterilized water 30-90ml, take the mode of solid state fermentation, under the condition of 30 ℃, cultivate 3-4 days; Product enzymic fermentation under the condition of 28-30 ℃, adds 250ml water mechanical shaking extraction solid culture 1-2h after cultivating and finishing, and centrifugal collection supernatant liquor is crude enzyme liquid;

Crude enzyme liquid is joined in xylo-oligosaccharide cooking liquor, and enzymatic hydrolysis condition is temperature: 40-50 ℃, pH=5.0-5.5; Enzyme concentration 5-10ml in 100ml cooking liquor, the xylo-oligosaccharide concentration in cooking liquor is 1.5%, after enzymolysis 1-2 days, obtains Xylose;

Or the above-mentioned high temperature steaming Xylose that calcium carbonate obtains (2) carries out neutralizing treatment and adjusts pH to 6.0-7.0, and Plate Filtration obtains supernatant liquor;

(4) add microorganism to remove the assorted sugar that comprises glucose and semi-lactosi in Xylose;

Selected microorganism is at least one of intestinal bacteria, yeast saccharomyces cerevisiae; The required nitrogenous source that ferments is at least one in yeast extract paste, yeast powder, corn steep liquor, Dried Corn Steep Liquor Powder, soybean-cake flour, ammonium sulfate, urea;

Above-mentioned Xylose, through after being concentrated into total sugar concentration and being 100-200g/L, adds above-mentioned nitrogenous source fully to dissolve, and the nitrogen concentration obtaining is 10-20g/L, is mixed with fermention medium;

At glucose concn, be 20-30/gL, yeast extract paste concentration is in the activation medium of 5-10g/L, to add at least one of intestinal bacteria, yeast saccharomyces cerevisiae and Angel Yeast, making at least one starting point concentration of intestinal bacteria, yeast saccharomyces cerevisiae and Angel Yeast is 1-2g/L, in the shaking table of 30 ℃, rotating speed activation 2-4h with 200-300rpm, centrifugal 15min abandons supernatant liquor, recentrifuge 10min after distilled water wash for the cell of bottom, collecting cell mud;

In fermention medium, add cell mud 10-20g/L, leavening temperature 25-35 ℃, fermentation rotating speed 200rpm, air flow is 1-1.5vv ^-1min ^-1, fermenting, fermentation time is 6-12h;

(5) centrifugal after fermentation, then gac or polymeric adsorbent, or both are in conjunction with Xylose is carried out to purification process;

(6) after wood sugar purifying, carry out concentration; Xylose Content in the Xylose obtaining is 700-800g/L;

(7) take the mode of gradient programmed cooling crystallization to obtain xylose product and xylose crystallization mother liquor;

(8) xylose crystallization mother liquor obtaining is separated through chromatographic system, obtain wood sugar phase and L-arabinose phase;

(9) wood sugar step (8) being obtained is back to mutually step (6) and merges concentrated with wood sugar refined solution, take the mode of gradient programmed cooling to carry out crystallization, obtain wood sugar crystal and xylose mother liquid, xylose mother liquid in xylose mother liquid and step (7) merges, enter to chromatographic fractionation system separation, repeating step (8); The L-arabinose that (8) are obtained obtains L-arabinose product and L-arabinose mother liquor through concentrated, crystallization mutually, and L-arabinose mother liquor is replied and utilized.

Wherein said agriculture and forestry organic waste material raw material comprises: corn cob, maize straw, rice straw, jowar stalk, cotton seed hulls, bagasse etc., the preferred cotton seed hulls of the present invention is raw material.

The pre-treating technology of the agriculture and forestry organic waste material raw materials such as cotton seed hulls that step (1) is described comprises: raw-material pulverizing, dedusting, washing three process.

The pretreatment technology preferred operations that the described agriculture and forestry organic waste material raw materials such as cotton seed hulls after pre-treatment are taked is easy, the simple infusion method of technique is processed, the soak solution that wherein said dipping pretreatment starting material are used is dilute sulphuric acid, dilute hydrochloric acid or rare oxalic acid, preferred rare oxalic acid soak solution wherein, reason is that in raw material, remaining oxalic acid can the mode with carbon source be utilized by Institute of Micro-biology in the fermenting process in later stage, adopt rare oxalic acid to carry out pretreated mode to raw material soaking and not only obtained good pre-treatment result, improved the wood sugar yield of high temperature steaming, also reduced the cost that the later stage adds the consumption of xylose metabolism and carbon source, and the difficulty of later stage desalting treatment.In rare oxalic acid soak solution used, the massfraction of oxalic acid is 0.1-0.5%, and soak solution is 4-8 with the massfraction ratio of raw material: 1, and the temperature of immersion is 30-100 ℃, the time of immersion is 6-24h.After immersion finishes, through the mode of Plate Filtration, immersion system is carried out to solid-liquid separation and obtain pretreated raw material and soak solution, soak solution is replied and is utilized, and the mode of isolated pretreated raw material by washing regulates raw material self pH value neutral to approaching.

Step (2) utilizes the method for the agriculture and forestry organic waste material raw material production xylo-oligosaccharide cooking liquors such as high-temperature cooking process hydrolysis of corncob, cotton seed hulls to be: the agriculture and forestry organic waste materials such as pretreated corn cob or cotton seed hulls are added in reaction unit, add 5-20 doubly to the water of solid material dry weight; Initial pH is 5.0-6.0; High temperature steaming 0.5-2h under the condition of 145 ℃-155 ℃; Mixing speed is 80-150rpm; Operating pressure is 0.5-1.0MPa.

Utilize the method for high-temperature cooking process hydrolysis agriculture and forestry organic waste material raw material production wood sugar cooking liquor to be: the agriculture and forestry organic waste material raw materials such as pretreated corn cob or cotton seed hulls to be added in reaction unit, to add 5-20 doubly to the water of solid material dry weight; At initial pH, be 4.5-6.0; High temperature steaming 1-3h under the condition of 160-180 ℃; Mixing speed is 80-150rpm; Operating pressure is 0.6-1.2MPa.In this operation, hemicellulose is by pyrohydrolysis, to be wood sugar under the environment of micro-sour partial neutral.

In the present invention, utilizing xylo-oligosaccharide yield in the agriculture and forestry organic waste material raw material production xylo-oligosaccharide cooking liquors such as high-temperature cooking process hydrolysis cotton seed hulls is 0.15-0.17g/g, and producing wood sugar yield in wood sugar cooking liquor is 0.20-0.22g/g.After two kinds of high temperature steaming hydrolytic processes, the steam of collect discharging is while hot to steam storage tank, can need thermal process that heat energy is provided for immersion treatment technique or other.This operation can be removed the impurity of a part of easy volatile in cooking liquor simultaneously, as furfural, phenol, acetic acid, also makes cooking liquor carry out concentrating to a certain degree.Cooling rear collection cooking liquor (containing hydrolysis residue) out carries out next step neutralizing treatment.

Step (3) utilize milk of lime to above-mentioned high temperature steaming xylo-oligosaccharide liquid carried out neutralizing treatment to pH at 8.0-9.0, again with phosphoric acid or hydrochloric acid readjustment pH to 6.0, Plate Filtration obtains supernatant liquor afterwards.Or the above-mentioned high temperature steaming Xylose that calcium carbonate obtains (2) carries out neutralizing treatment and adjusts pH to 6.0-7.0, and Plate Filtration obtains supernatant liquor;

The present invention utilizes milk of lime high temperature steaming xylo-oligosaccharide liquid to carry out neutralizing treatment, regulates boiling mixed solution to pH at 8.0-9.0, afterwards again with phosphoric acid or hydrochloric acid readjustment pH to 6.0 left and right.The inhibition of the some effects enzyme liquid enzymolysis efficiency of taking neutralizing treatment cooking mixture also can remove to exist in high temperature steaming xylo-oligosaccharide liquid.Owing to having organic toxin and the metal ions such as a small amount of furfural, phenol, acetic acid, pigment in xylo-oligosaccharide cooking liquor, crude enzyme liquid that next step aspergillus niger produces is carried out to enzymolysis acquisition Xylose and have larger impact.Therefore the present invention first adopt milk of lime cross in and xylo-oligosaccharide high temperature steaming liquid to pH at 8.0-9.0, acid in now can not only be completely and in cooking liquor, and the impurity in the calcium sulfate flocks energy effective elimination xylo-oligosaccharide liquid producing, to realize the raising of enzymolysis efficiency.Afterwards again with phosphoric acid or hydrochloric acid readjustment pH to 6.0 left and right, for producing enzyme liquid, aspergillus niger provides suitable pH environment when the enzymolysis.After this neutralizing treatment, the wood sugar yield of xylo-oligosaccharide cooking liquor in enzymolysis process can be increased to more than 80%, reaches the requirement of producing Xylose.

The present invention utilizes calcium carbonate to carry out neutralizing treatment to high temperature steaming Xylose, regulate boiling mixed solution to pH at 6.0-7.0.The method is not only conducive to the pH environment that later stage purifying concentration is suitable, and the calcium sulfate flocks of generation also can be removed the impurity existing in cooking liquor.

The throw out elimination in the lump that adopts plate filter that the cotton seed hulls waste residue after high temperature steaming and neutralization are produced, obtains supernatant liquor and is the cooking liquor after neutralizing treatment.Concrete grammar is: by charging pump, the cooking mixture after neutralization is pressed into filter chamber, has filtered rear concentrated discharge, then add 30-50 ℃ of water 2L, solid is washed to repeated washing 2 times.

Xylo-oligosaccharide cooking liquor after neutralizing treatment, adds the crude enzyme liquid that the preferred aspergillus niger 2349 of aspergillus niger produces to carry out enzymolysis production wood sugar to the xylo-oligosaccharide in cooking liquor, to obtain wood sugar enzymolysis solution.Relatively still less, and the further enzymolysis of xylo-oligosaccharide completely that makes not degrade in high temperature steaming liquid obtains wood sugar to the power consumption of this technique, improved raw material production wood sugar yield.The crude enzyme liquid that this process is used, for self-control, has been saved Financial cost and Environmental costs.

Xylo-oligosaccharide cooking liquor after the crude enzyme liquid that the aspergillus niger 2349 of employing high yield zytase and xylosidase produces is processed (3) carries out enzymolysis and obtains wood sugar enzymolysis solution.Shake-flask seed substratum: Cha Shi substratum (SODIUMNITRATE 3g, dipotassium hydrogen phosphate 1g, magnesium sulfate 0.5g, Repone K 0.5g, ferrous sulfate 0.01g, sucrose 30g distilled water 1000ml), 115 ℃ of sterilizing 30min, access aspergillus niger 2349 is cultivated and is had spore shape form to be advisable to the aspergillus niger length in seed culture medium in 3-4 days under the condition of 28-30 ℃.Producing enzymic fermentation substratum is: sucrose 2.4g, MgSO ₄0.12g, K ₂hPO ₄0.12g, KH ₂pO ₄0.24g, FeSO ₄0.0012g, NaNO ₃0.012g, uses 100ml water dissolution, mixes with the mixture of wheat bran with corn cob, and the mixture of corn cob and wheat bran amounts to 100g, and corn cob and wheat bran mass ratio are 3: 7-7: 3, and sterilizing 30min under the condition of 115 ℃.Under the condition of aseptic technique, in 100g fermention medium, add the aspergillus niger seed liquor 10-15ml with spore shape form, add sterilized water 30-90ml, take the mode of solid state fermentation, under the condition of 30 ℃, cultivate 3-4 days.Product enzymic fermentation under the condition of 28-30 ℃, adds 250ml water mechanical shaking extraction solid culture 1-2h after cultivating and finishing, and centrifugal collection supernatant liquor is crude enzyme liquid.Crude enzyme liquid is joined in xylo-oligosaccharide cooking liquor, and enzymatic hydrolysis condition is temperature: 40-50 ℃, pH=5.0-5.5.Enzyme concentration 5-10ml in 100ml cooking liquor, the xylo-oligosaccharide concentration in cooking liquor is 1.5%, after enzymolysis 1-2 days, obtains wood sugar enzymolysis solution.

Step (4) adds microorganism to remove the assorted sugar that comprises glucose and semi-lactosi in Xylose, and wood sugar and the L-arabinose effective constituent in fermenting xylose liquid not, to remove the interference of the assorted sugar that affects wood sugar and the two fractional crystallization of L-arabinose.The present invention when utilizing microbe fermentation method to process, bacterial classification used is intestinal bacteria, yeast saccharomyces cerevisiae at least one, preferred Angel Yeast; The required nitrogenous source that ferments is at least one in yeast extract paste, yeast powder, corn steep liquor, Dried Corn Steep Liquor Powder, soybean-cake flour, ammonium sulfate, urea, preferred yeast powder or Dried Corn Steep Liquor Powder.

Xylose, after MVR machinery recompression high-efficiency evaporating system is concentrated into total sugar concentration and is 100-200g/L, adds above-mentioned nitrogenous source fully to dissolve, and the nitrogen concentration obtaining is 10-20g/L, is mixed with fermention medium.At glucose concn, be 20-30g/L, yeast extract paste concentration is to add Angel Yeast bacterial strain in the activation medium of 5-10g/L, making its starting point concentration is 1-2g/L, in the shaking table of 30 ℃, rotating speed activation 2-4h with 200-300rpm, centrifugal 15min abandons supernatant liquor, recentrifuge 10min after distilled water wash for the cell of bottom, collecting cell mud.In fermention medium, add yeast slurry 10-20g/L in fermentor tank, leavening temperature 25-35 ℃, fermentation rotating speed 200rpm, air flow 1-1.5vv ^-1m ^-1, fermentation volume 16L, ferments, and fermentation time is 6-12h.

Step in the present invention (5) is the Xylose after Angel Yeast is processed, and takes centrifugal mode to remove thalline and the insoluble substance in treatment solution (4), obtains wood sugar treatment solution.Isolated somatic cells adds after the activation medium of fresh sterile, and the activation culture process through in step (4), proceeds in next batch fermention medium and reuse, and many batches of formulas move in circles, until apoptosis.

The wood sugar treatment solution obtaining adopts the mode of gac or macroporous adsorbent resin or both combinations to carry out purification process to it.The huge active surface that Powdered Activated Carbon has, absorption impurity and pigment, then remove the impurity of absorption together with gac in the lump by filtering, and reaches the refining object of liquid glucose.The process of charcoal absorption impurity belongs to physical adsorption, and the organic ability of charcoal absorption is much larger than absorption inorganic salts, and the ability of absorption macromole organic pigment is much larger than the ability of absorption small molecules organic pigment.Ion-exchange: liquid glucose by cationic exchange and anionresin after, impurity cationic in liquid glucose and impurity negatively charged ion are all adsorbed on ion exchange resin and are removed, and these foreign ions are the integral part of sulfuric acid in liquid glucose, organic acid and the grey impurity that grades.From resin, submit the hydrogen ion and the hydroxide ion that change to liquid glucose and be combined into water.Wherein a kind of mode of activated carbon treatment is, to according to being 10-30% with wood sugar mass ratio, ratio adds gac in hydrolyzed solution, at 30-90 ℃, slowly stirs, and pigment, nitrogenous thing, organism and colloid in absorption boiling liquid, 20min is to 2h in decolouring.Xylose after activated carbon treatment is removed gac part through the mode of Plate Filtration, and the xylose hydrolysis fluid that obtains achromaticity and clarification is the Xylose after activated carbon treatment.

Xylose after activated carbon treatment adopts the processing mode of zwitterion resins exchange to remove the ion in Xylose, and the Zeo-karb trade mark of employing is 732, is strongly acidic styrene type cation exchange resin, and exchange capacity is 4.5Mmol/g; The anionite-exchange resin trade mark adopting is that D201 is strong-basicity styrene series anion exchange resin, and exchange capacity is 3.7Mmol/g.732 strongly acidic styrene type cation exchange resins and D201 strong-basicity styrene series anion exchange resin are successively connected hydrolyzed solution are carried out to desalting treatment.Resin column blade diameter length ratio is 1: 4-5, the material of resin column is glass reinforced plastic or carbon steel plastics, and inlet amount is a 1.2-1.6 per hour column volume, and working temperature is room temperature, the electric conductivity value of on-line monitoring discharging feed liquid, the specific conductivity < 50 μ s/cm of assurance discharging feed liquid.Long-term use even loses ion-exchange capacity reduction, need according to circumstances to resin, carry out activation treatment.Activation method can be determined according to pollution condition and condition, and general positive resin is subject to Fe in softening ³⁺pollute progressively dilution after acceptable salts acid soak.Negative resin is subject to Organic pollutants, and available 10%NaCl+2-5%NaOH mixing solutions soaks or drip washing.

The Xylose Content that Xylose after the purifying that step (6) obtains (5) carries out in Xylose that certain concentration obtains is 700-800g/L.The concentration technique of Xylose is divided into wherein a kind of or be combined with between two in membrane concentration and MVR machinery recompression concentration technology.Membrane concentration operation comprises ultrafiltration, nanofiltration, reverse osmosis three process.Ultrafiltration is by hollow-fibre ultrafiltration device, to remove in cooking liquor macromolecular impurity as protein, pigment etc.Nanofiltration is concentrated is to utilize suitable nanofiltration membrane cooking liquor to be carried out to selectivity is concentrated also has partially desalted effect concurrently simultaneously, to alleviate the burden of postorder ion-exchange.From the viewpoint of energy consumption, in technique, evaporation concentration energy consumption is very large, and a large amount of energy consumptions is also brought environmental pollution, and the concentrated energy consumption that can greatly reduce evaporation of nanofiltration, shortens the time of evaporating, and improves product quality.Reverse osmosis is nanofiltration to be seen through to liquid carry out desalination recovery, replys utilization, thereby save water resources to obtain pure water.Adopt MVR machinery recompression high-efficiency evaporating system concentration technology high temperature steaming liquid.Whole plant energy-saving effect is outstanding, except start up, in whole evaporative process without raw steam.First the steam of low-temperature level compresses through compressor, and temperature, pressure improve, and heat content increases, and then enters interchanger condensation, to make full use of the latent heat of steam.First technique be pumped into high temperature steaming liquid in falling-film evaporator, by Matter Transfer pump at heating tube internal recycle.Initial steam is given heat outside pipe with live steam, solution ebuillition of heated is produced to secondary vapour, and the secondary vapour of generation is sucked by turbine boosted fan, and after supercharging, secondary stripping temperature improves, and as heat source, enters heating chamber circulating and evaporating.After normal startup, turbo-driven compressor sucks secondary steam, becomes heating steam after supercharging, carries out continually circulating and evaporating like this.The moisture evaporating finally becomes water of condensation and discharges.

Wood sugar concentrated solution after step (7) is processed (6), adopts the mode of gradient programmed cooling crystallization to obtain xylose product and xylose crystallization mother liquor; To be concentrated into the Xylose that total sugar concentration is 700-800g/L, adopt the mode crystallization of gradient programmed cooling to go out wood sugar crystal, obtain xylose crystallization mother liquor simultaneously.It is 60-80 ℃ that the mode of gradient programmed cooling is taked initial temperature, final temperature is 20-30 ℃, the mode of 0.5-2 per hour ℃ is taked in cooling, in the time of 45-55 ℃, add solid content massfraction be 0.1% commodity wood sugar as crystal seed, and under final temperature growing the grain 12-24h.Owing to having passed through microbiological treatment before Xylose, the interference of assorted sugar to later stage product crystallization such as glucose, semi-lactosi have been removed, its crystallization yield is compared and has been improved nearly 10% with traditional product crystallization yield that contains the assorted sugar such as glucose, semi-lactosi, reached 70% left and right, the purity of crystal is greater than 99%.Crystallisation process finishes by centrifugation, obtains xylose mother liquid, the dry crystalline xylose product that obtains after washing with alcohol for crystal.

Wood sugar and L-arabinose in the chromatographic system separating xylose mother liquor utilizing in described step (8), obtain wood sugar phase and select the simulated moving bed chromatography of the Zeo-karb of the calcium type that is filled with, plumbous type or sodium type that level of automation is high to come separated with the method for L-arabinose phase.Because simulated moving bed chromatography isolation technique is carried out the separated of wood sugar and L-arabinose to xylose crystallization mother liquor, whole automation technolo degree is high, simple to operate, has greatly saved the consumption of resin, has reduced pollution.

The xylose crystallization mother liquor that step (7) is obtained enters the pillar that 20 2.5*50cm are housed, and the simulated moving bed chromatography system that to have filled calcium type resin cation (R.C.) be sorbent material, carries out chromatographic separation.Separation temperature 60-85 ℃, xylose mother liquid charging feeding rate 300-500mL/h, pure water elution speed 1-2L/h.After system balancing, collect wood sugar and L-arabinose two-phase liquid, the purity of two-phase is all more than 95%, and it is more than 90% replying yield; After simulated moving bed chromatography system separation, obtain wood sugar phase and L-arabinose phase two-phase liquid.

The wood sugar that described step (9) obtains (8) is back to mutually step (6) and merges concentrated with wood sugar refined solution, take the mode of gradient programmed cooling to carry out crystallization, obtain wood sugar crystal and xylose mother liquid, xylose mother liquid in xylose mother liquid and step (7) merges, enter to chromatographic fractionation system separation, repeating step (8); The L-arabinose that (8) are obtained obtains L-arabinose product and L-arabinose mother liquor through concentrated, crystallization mutually, and L-arabinose mother liquor is replied and utilized.

Advantage of the present invention

1, the inventive method be take agriculture and forestry organic waste material as raw material, through simple and practical pretreatment technology in conjunction with the high temperature steaming of advanced clean and effective or boiling the method in conjunction with enzymolysis, the technique of agriculture and forestry organic waste material raw material production xylose hydrolysis fluid is substituted to classical acid hydrolysis process, whole simple in production process operation safety, corrodibility to equipment is little, avoided a large amount of acid in traditional hydrolysis process, the consumption of alkali, and the system obtaining after processing is without neutralizing and producing without waste residue, the quantity discharged of the sewage in hydrolytic process has obtained significantly reducing, 1 ton of wood sugar quantity of wastewater effluent of every production is less than 50 tons.The xylose hydrolysis fluid making with traditional sulfuric acid process is compared saltiness and has been reduced by 80%.Two kinds of hydrolyzed solutions that the present invention obtains are after microorganism and purification process, crystallization yield and the purity of wood sugar and L-arabinose product are greatly improved, in the xylose mother liquid simultaneously obtaining, also only contain utilization that the assorted sugar of wood sugar, L-arabinose and minute quantity is more conducive to the later stage with separated, avoided the waste of raw material, raw material has been obtained more fully and the utilization of science.The present invention produces raw materials used wide material sources, the compact easy handling of production technique, and the high wood sugar of added value of product that can simultaneously obtain and two kinds of products of L-arabinose, whole production technique is clean and effective both, has again sustainability, meets the idea of biorefinery.

2, the present invention is by choosing specific microbial strains, Xylose after the purified processing of specific consumption, remove its assorted sugar that affects later stage wood sugar and L-arabinose purified crystals process as glucose, semi-lactosi etc., do not lose wood sugar and L-arabinose, the separation of being convenient to the later stage is produced simultaneously.Effectively improve and avoided in old technology due to the assorted sugared excessive concentration in xylose mother liquid, osmotic pressure is excessive, impact for assorted sugar such as thalli growth and removal glucose is larger, also needs mother liquor Macrodilution in fermenting process, has increased the weight of to process the drawback of load.Therefore the present invention takes to utilize yeast to process the assorted sugar such as glucose in xylose hydrolysis fluid before xylose crystalline.Because sugared concentration in xylose hydrolysis fluid is lower, osmotic pressure is suitable, and the assorted sugar such as glucose that yeast can fully consume are wherein grown, so the clearance of the assorted sugar such as glucose and semi-lactosi is high.Xylose has passed through microbiological treatment, in the Xylose obtaining, also only contain the assorted sugar of wood sugar, L-arabinose and minute quantity, avoided the interference of assorted sugar to later stage wood sugar and the crystallization of L-arabinose product such as glucose, semi-lactosi, the crystallization yield of later stage wood sugar and L-arabinose and purity are greatly improved.Compare and improved nearly 10 percentage point with the yield of the traditional hydrolyzed solution crystalline xylose of not removing the assorted sugar such as glucose, semi-lactosi, reached 70% left and right, the purity of wood sugar crystal is greater than 99%.

3, because the assorted sugar in early stage hydrolyzed solution has obtained effective removal, other sugar that only contain wood sugar, L-arabinose and minute quantity in xylose crystallization mother liquor, the purity of wood sugar, L-arabinose effective constituent with do not take microbial method to process to compare to have obtained obvious rising, xylose mother liquid can be more effectively for the reply utilization in later stage, raw material has been obtained more fully and the utilization of science, significantly improved the utility value of xylose mother liquid.In conjunction with simulated moving bed chromatography isolation technique, xylose crystallization mother liquor is carried out to the separated of wood sugar and L-arabinose, whole automation technolo degree is high, simple to operate, has greatly saved the consumption of resin, has reduced pollution.Because the purity of two kinds of products in mixed solution is high, product separation effect and yield have significantly been improved.The L-arabinose product with high added value extracting from xylose mother liquid, effectively reduces the production cost of L-arabinose raw material, has significantly reduced the cost of whole technique, has avoided the waste of raw material, has significantly improved economic worth.

The problems such as 4, concentration process energy consumption is high, quantity of wastewater effluent is large, the present invention has developed a set of novel concentrated rectification process-MVR machinery recompression evaporation process, realizes zero release and the high yield of waste water.MVR is the energy that re-uses the secondary steam that it self produces, thereby reduces a technology of the demand of the energy to external world.Its working process is that the steam of low-temperature level compresses through compressor, and temperature, pressure improve, and heat content increases, and then enters interchanger condensation, to make full use of the latent heat of steam.Except opening starting, in whole evaporative process without raw steam, energy-conserving and environment-protective.The MVR machinery recompression evaporation process that the present invention adopts replaces traditional multiple-effect evaporation concentration technique, can realize the recycling of water and the significantly reduction of energy consumption, and evaporating water per ton only needs power consumption 20 degree, Energy Intensity Reduction more than 80%.The application of this technology simultaneously has low working cost, operate steadily and feature that level of automation is high.

5, the present invention produces raw materials used wide material sources, and production technique is compact, easy handling, can obtain two kinds of high value added products of wood sugar and L-arabinose simultaneously, improves greatly the added value of agricultural-food.Effectively solve the high pollution existing in existing industrial production wood sugar process, high energy consumption, the not high and larger problem of production cost of the rate of recovery.Whole production technique is clean and effective both, has again sustainability, meets the idea of biorefinery, and production cost is controlled in the 60-70% of productive value simultaneously, has larger using value.

Accompanying drawing explanation

Schema of the present invention.

Embodiment

Embodiment 1

(1) pre-treatment is prepared the method for Xylose in conjunction with high temperature steaming or boiling in conjunction with enzymolysis

Selecting agriculture and forestry organic waste material cotton seed hulls is raw material.Cotton seed hulls pre-treatment comprises pulverizing, the dedusting of raw material, the three process of washing.First order number is pulverized and be sieved to cotton seed hulls purchase being obtained in pulverizer is 10 object solid particulates.Then the impurity in cotton seed hulls particle and upper layer silt dust etc. is carried out to dust removal process, to alleviate rear operation burden.Finally the agriculture and forestry organic waste material particles such as cotton seed hulls after dust removal process are washed to protein, pigment, pectin substance, the ash of removing part in its fiber, the impurity such as tannin teas.The three-procedure of pre-treatment is the quality that improves later stage product, the important measures that reduce costs.

Cotton seed hulls after pre-treatment carries out pre-treatment.Pretreatment technology selection operation is easy, the simple infusion method of technique is processed, the soak solution that pre-treatment cotton seed hulls is used for massfraction be 0.1% dilute sulphuric acid, immersion liquid used is 5: 1 with the massfraction ratio of raw material, soaking temperature is 60 ℃, soak time is 6h.After immersion finishes, through the mode of Plate Filtration, immersion system is carried out to solid-liquid separation and obtain pretreated raw material and soak solution, soak solution is replied and is utilized, and the mode of isolated pretreated raw material by washing regulates raw material self pH value neutral to approaching.

(2) high-temperature cooking process is produced and be take wood sugar as main high temperature steaming Xylose

Through pre-treatment and pretreated particle cotton seed hulls 3Kg (dry weight), add in pyroreaction still, high temperature stainless steel reactor wherein, capacity 50L, adds 10 times to the water of solid material dry weight.Anchor formula is adjustable, and rotating speed stirs, and electrically heated is cooling.Enclosed high pressure heating, Temperature Setting is 165 ℃, and 70rpm stirs, and operating pressure is 1.0MPa left and right, and maintains 2h at 165 ℃, and the time of intensification and cooling of controlling is within 0.5h.After boiling hydrolysis, high-temperature steam is expelled to storage tank from high temperature conduit; In cooking liquor, the productive rate of wood sugar is 0.20-0.22g/g, and xylo-oligosaccharide productive rate is 0.03-0.04g/g, and glucose productive rate is that 0.02-0.3g/g and L-arabinose productive rate are 0.01-0.02g/g (with cotton seed hulls dry weight basis).

Utilize calcium carbonate to carry out neutralizing treatment to high temperature steaming Xylose, adjusting boiling mixed solution is to pH 6.0, and the method is not only conducive to the pH environment that later stage purifying concentration is suitable, and the calcium sulfate flocks of generation also can be removed the impurity existing in cooking liquor.The throw out elimination in the lump that adopts plate filter that the cotton seed hulls waste residue after high temperature steaming and neutralization are produced, obtains supernatant liquor and is the Xylose after neutralizing treatment.Concrete grammar is: by charging pump, the cooking mixture after neutralization is pressed into filter chamber, has filtered rear concentrated discharge, then add 50 ℃ of water 5L, solid is washed, repeated washing 2 times, collects filtrate and obtain Xylose.

(3) purifying of Xylose and concentration

It is 120g/l that Xylose is concentrated into total sugar concentration through MVR machinery recompression high-efficiency evaporating system.Now add 12g/l Dried Corn Steep Liquor Powder to be configured to fermention medium, be driven in 20L fermentor tank stand-by.At glucose concn, be 20g/L, yeast extract paste concentration is to add Angel Yeast in the substratum of 5g/L, making its starting point concentration is 1g/L, in the shaking table of 30 ℃, rotating speed activation 2h with 200rpm, centrifugal 15min abandons supernatant liquor, and recentrifuge 10min after distilled water wash for the yeast of bottom collects yeast slurry.

In fermention medium, add yeast slurry 20g/L in fermentor tank, 30 ℃ of leavening temperatures, fermentation rotating speed 200rpm, air flow 1vv ^-1m ^-1, fermentation volume 16L, ferments.Between yeast phase, timing sampling HPLC analyzes the wherein content of each sugar component, after fermentation 6h, only contains the ethanol that wood sugar, L-arabinose and glucose metabolism are produced in fermented liquid, and the clearance of the assorted sugar such as glucose, more than 98%, now stops fermentation.Xylose after yeast saccharomyces cerevisiae ATCC9080 processes, take centrifugal mode to remove thalline and the insoluble substance in treatment solution, obtain wood sugar treatment solution, isolated somatic cells adds after the activation medium of fresh sterile, through above-mentioned activation culture process, proceed in next batch fermention medium and reuse, many batches of formulas move in circles, until apoptosis.

By 65L wood sugar treatment solution (wood sugar 20-22g/l, L-arabinose 2-3g/l) be driven in the bleacher of 100L, to according to being 15% with wood sugar mass ratio, ratio adds gac in hydrolyzed solution, at 60 ℃, slowly stir decolouring 1h, pigment, nitrogenous thing, organism and colloid in absorption boiling liquid.Xylose after activated carbon treatment is removed gac part through the mode of Plate Filtration, and the xylose hydrolysis fluid that obtains achromaticity and clarification is the Xylose after activated carbon treatment.

Xylose after activated carbon treatment adopts the processing mode of zwitterion resins exchange to remove the ion in Xylose.Xylose is passed through to 001*7 type Zeo-karb series connection 201*7 type anionite-exchange resin, at flow velocity 1L/h, under the condition of room temperature, the electric conductivity value of on-line monitoring discharging feed liquid, the specific conductivity < 50 μ s/cm of assurance discharging feed liquid.Xylose after gac and ion-exchange resin purification, takes the mode of MVR machinery recompression high-efficiency evaporating to carry out concentration, and the total sugar concentration to Xylose is 800g/L.

(4) the separated and crystallization processes of wood sugar and L-arabinose

After Xylose is purified and concentrated, its initial total sugar mass concentration is 800g/L, and the mass concentration that now contains wood sugar in hydrolyzed solution is 720g/L, and the mass concentration that contains L-arabinose is 80g/L.

What carry out that the crystallization mode of wood sugar takes is the method for gradient programmed cooling, initial temperature is 70 ℃, according to the rate of temperature fall crystallisation by cooling of 2 ℃/h, when temperature is down to 50 ℃, add 0.25% crystal seed, crystal seed is educated after brilliant 1h, then the rate of temperature fall according to 1.5 ℃/h continues crystallisation by cooling, be cooled to 20 ℃ of left and right, growing the grain 6h, it is centrifugal by supernatant liquor and the concentrated fore portion merging of Xylose after the wood sugar crystal of separating out is taken out, repeat above-mentioned condensing crystal process, the centrifugal solid-phase obtaining rinses 2 times with dehydrated alcohol, and then 45 ℃ of drying in oven obtain xylose crystalline product.Crystallization finishes rear acquisition wood sugar crystal and xylose crystallization mother liquor, and wherein the purity of xylose product is more than 99%, and crystallization yields is 67%.In xylose crystallization mother liquor, contain the wood sugar of 210g/L and the L-arabinose of 73g/L.

The xylose crystallization mother liquor of above-mentioned acquisition is entered to the pillar that 20 2.5*50cm are housed, the simulated moving bed chromatography system that to have filled calcium type resin cation (R.C.) be sorbent material, carries out chromatographic separation.65 ℃ of separation temperatures, feeding rate 200mL/h, pure water elution speed 1.2L/h.After system balancing, collect wood sugar and L-arabinose two-phase liquid, the purity of two-phase is all more than 95%, and it is more than 98% replying yield.

It is 800g/L that the L-arabinose that simulation moving-bed separation is obtained adopts MVR machinery recompression high-efficiency evaporating system to be concentrated into L-arabinose concentration at 80 ℃, then according to the rate of temperature fall crystallisation by cooling of 2.5 ℃/h, when temperature is down to 50 ℃, add 0.25% crystal seed, crystal seed is educated after brilliant 1h, then the rate of temperature fall according to 1.5 ℃/h continues crystallisation by cooling, be cooled to 20 ℃ of left and right, growing the grain 6h, by centrifugal after the L-arabinose crystal taking-up of separating out, the isolated L-arabinose of supernatant liquor and chromatographic system is merged mutually, repeat above-mentioned condensing crystal process, the centrifugal solid-phase obtaining rinses 2 times with dehydrated alcohol, and then 45 ℃ of drying in oven, L-arabinose crystallization yields is 68%, purity is more than 99%.

The wood sugar that xylose mother liquid separation is obtained returns mutually and is incorporated in the initial Xylose of the 4th step, adopting MVR machinery recompression high-efficiency evaporating system to be concentrated into xylose concentration is 800g/L, repeat xylose crystalline step, obtain wood sugar crystal and xylose crystallization mother liquor, xylose crystalline productive rate is 60%, purity is more than 99%, after the xylose mother liquid that the xylose mother liquid xylose crystalline initial with the 4th step that this step obtains obtains mixes, enter simulation moving-bed separation system and obtain wood sugar phase and L-arabinose phase, with this repeatedly.

Embodiment 2

(1) pre-treatment is prepared the method for Xylose in conjunction with high temperature steaming or boiling in conjunction with enzymolysis

Selecting agriculture and forestry organic waste material cotton seed hulls is raw material.Cotton seed hulls pre-treatment comprises pulverizing, the dedusting of raw material, the three process of washing.First order number is pulverized and be sieved to cotton seed hulls purchase being obtained in pulverizer is 20 object solid particulates.Then the impurity in cotton seed hulls particle and upper layer silt dust etc. is carried out to dust removal process, to alleviate rear operation burden.Finally the agriculture and forestry organic waste material particles such as cotton seed hulls after dust removal process are washed to protein, pigment, pectin substance, the ash of removing part in its fiber, the impurity such as tannin teas.The three-procedure of pre-treatment is the quality that improves later stage product, the important measures that reduce costs.

Cotton seed hulls after pre-treatment carries out pre-treatment.Pretreatment technology selection operation is easy, the simple infusion method of technique is processed, the soak solution that pre-treatment cotton seed hulls is used for massfraction be 0.25% dilute sulphuric acid, immersion liquid used is 6: 1 with the massfraction ratio of raw material, and soaking temperature is 30 ℃, and soak time is 24h.After immersion finishes, through the mode of Plate Filtration, immersion system is carried out to solid-liquid separation and obtain pretreated raw material and soak solution, soak solution is replied and is utilized, and the mode of isolated pretreated raw material by washing regulates raw material self pH value neutral to approaching.

(2) high-temperature cooking process production high temperature steaming xylo-oligosaccharide liquid is prepared Xylose in conjunction with enzymolysis

Through pre-treatment and pretreated particle cotton seed hulls 4.5Kg (dry weight), add in pyroreaction still, high temperature stainless steel reactor wherein, capacity 50L, adds 8 times to the water of solid material dry weight.Anchor formula is adjustable, and rotating speed stirs, and electrically heated is cooling.Enclosed high pressure heating, Temperature Setting is 155 ℃, and 70rpm stirs, and operating pressure is 0.8MPa left and right, and maintains 1h at 155 ℃, and the time of intensification and cooling of controlling is within 0.5h.After boiling hydrolysis, high-temperature steam is expelled to storage tank from high temperature conduit; In cooking liquor, the productive rate of xylo-oligosaccharide is 0.14g/g, and xylose yield is 0.06g/gl, and glucose productive rate is that 0.021g/g and L-arabinose productive rate are 0.013g/g (with cotton seed hulls dry weight basis).

The present invention utilizes milk of lime high temperature steaming xylo-oligosaccharide liquid to carry out neutralizing treatment, regulate boiling mixed solution to pH 8.5, acid in now can not only be completely and in cooking liquor, and the impurity in the calcium sulfate flocks energy effective elimination xylo-oligosaccharide liquid producing, to realize the raising of enzymolysis efficiency.Afterwards again with phosphoric acid or hydrochloric acid readjustment pH to 6.0 left and right, for producing enzyme liquid, aspergillus niger provides suitable pH environment when the enzymolysis.After this neutralizing treatment, the wood sugar yield of xylo-oligosaccharide cooking liquor in enzymolysis process can be increased to more than 80%, reaches the requirement of producing Xylose.The throw out elimination in the lump that adopts plate filter that the cotton seed hulls waste residue after high temperature steaming and neutralization are produced, obtains supernatant liquor and is the xylo-oligosaccharide cooking liquor after neutralizing treatment.Concrete grammar is: by charging pump, the cooking mixture after neutralization is pressed into filter chamber, has filtered rear concentrated discharge, then add 50 ℃ of water 5L, solid is washed, repeated washing 2 times, collects filtrate and obtain xylo-oligosaccharide cooking liquor.

Xylo-oligosaccharide cooking liquor after the crude enzyme liquid that the aspergillus niger 2349 (being purchased from Chinese industrial microbial strains preservation administrative center) of employing high yield zytase and xylosidase produces is processed (3) carries out enzymolysis and obtains wood sugar enzymolysis solution.Shake-flask seed substratum: Cha Shi substratum, 115 ℃ of sterilizing 30min, access aspergillus niger 2349 is cultivated and is had spore shape form to be advisable to the aspergillus niger length in seed culture medium in 4 days under the condition of 28 ℃.Producing enzymic fermentation substratum is: sucrose 2.4g, MgSO ₄0.12g, K ₂hPO ₄0.12g, KH ₂pO ₄0.24g, FeSO ₄0.0012g, NaNO ₃0.012g, with mixing with the mixture of wheat bran with corn cob after 100ml water dissolution, the mixture of corn cob and wheat bran amounts to 100g, and corn cob and wheat bran mass ratio are 6: 4, sterilizing 30min under the condition of 115 ℃.Under the condition of aseptic technique, in 100g fermention medium, add the aspergillus niger seed liquor 15ml with spore shape form, add sterilized water 60ml, take the mode of solid state fermentation, under the condition of 30 ℃, cultivate 4 days.Product enzymic fermentation under the condition of 28 ℃, adds 250ml water mechanical shaking extraction solid culture 2h after cultivating and finishing, and centrifugal collection supernatant liquor is crude enzyme liquid.Crude enzyme liquid is joined in xylo-oligosaccharide cooking liquor, and enzymatic hydrolysis condition is temperature: 50 ℃, and pH=5.5.Enzyme concentration 10ml in 100ml cooking liquor, the xylo-oligosaccharide concentration in cooking liquor is 1.5%, through enzymolysis, after 2 days, more than 80% xylo-oligosaccharide is converted into wood sugar, obtains Xylose.

(3) purifying of Xylose and concentration

It is 150g/l that Xylose is concentrated into total sugar concentration through MVR machinery recompression high-efficiency evaporating system.Now add 20g/l Dried Corn Steep Liquor Powder to be configured to fermention medium, be driven in 20L fermentor tank stand-by.At glucose concn, be 30g/L, yeast extract paste concentration is to add Angel high activity dried yeast in the substratum of 10g/L, be purchased from supermarket, Angel company produces, and making its starting point concentration is 2g/L, in the shaking table of 30 ℃, rotating speed activation 4h with 300rpm, centrifugal 15min abandons supernatant liquor, and recentrifuge 10min after distilled water wash for the yeast of bottom collects yeast slurry.

In fermention medium, add yeast slurry 10g/L in fermentor tank, 32 ℃ of leavening temperatures, fermentation rotating speed 200rpm, air flow 1.4vv ^-1m ^-1, fermentation volume 16L, ferments.Between yeast phase, timing sampling HPLC analyzes the wherein content of each sugar component, after fermentation 12h, only contains the ethanol that wood sugar, L-arabinose and glucose metabolism are produced in fermented liquid, and the clearance of the assorted sugar such as glucose, more than 98%, now stops fermentation.Xylose after Angel high activity dried yeast is processed, take centrifugal mode to remove thalline and the insoluble substance in treatment solution, obtain wood sugar treatment solution, isolated somatic cells adds after the activation medium of fresh sterile, through above-mentioned activation culture process, proceed in next batch fermention medium and reuse, many batches of formulas move in circles, until apoptosis.

By 65L wood sugar treatment solution (wood sugar 20-22g/l, L-arabinose 2-3g/l) be driven in the bleacher of 100L, to according to being 25% with wood sugar mass ratio, ratio adds gac in hydrolyzed solution, at 30 ℃, slowly stir decolouring 40min, pigment, nitrogenous thing, organism and colloid in absorption boiling liquid.Xylose after activated carbon treatment is removed gac part through the mode of Plate Filtration, and the xylose hydrolysis fluid that obtains achromaticity and clarification is the Xylose after activated carbon treatment.

Xylose after activated carbon treatment adopts the processing mode of zwitterion resins exchange to remove the ion in Xylose.Xylose is passed through to 001*7 type Zeo-karb series connection 201*7 type anionite-exchange resin, at flow velocity 1L/h, under the condition of room temperature, the electric conductivity value of on-line monitoring discharging feed liquid, the specific conductivity < 50 μ s/cm of assurance discharging feed liquid.Xylose after gac and ion-exchange resin purification, takes the mode of MVR machinery recompression high-efficiency evaporating to carry out concentration, and the total sugar concentration to Xylose is 700g/L.

(4) the separated and crystallization processes of wood sugar and L-arabinose

After Xylose is purified and concentrated, its initial total sugar mass concentration is 700g/L, and the mass concentration that now contains wood sugar in hydrolyzed solution is 630g/L, and the mass concentration that contains L-arabinose is 70g/L.

What carry out that the crystallization mode of wood sugar takes is the method for gradient programmed cooling, initial temperature is 80 ℃, according to the rate of temperature fall crystallisation by cooling of 3 ℃/h, when temperature is down to 50 ℃, add 0.1% crystal seed, crystal seed is educated after brilliant 2h, then the rate of temperature fall according to 2 ℃/h continues crystallisation by cooling, be cooled to 30 ℃ of left and right, it is centrifugal by supernatant liquor and the concentrated fore portion merging of Xylose after the wood sugar crystal of separating out is taken out, repeat above-mentioned condensing crystal process, the centrifugal solid-phase obtaining rinses 2 times with dehydrated alcohol, and then 45 ℃ of drying in oven obtain xylose crystalline product.Crystallization finishes rear acquisition wood sugar crystal and xylose crystallization mother liquor, and wherein the purity of xylose product is more than 99%, and crystallization yields is 65%.In xylose crystallization mother liquor, contain the wood sugar of 200g/L and the L-arabinose of 70g/L.

The xylose crystallization mother liquor of above-mentioned acquisition is entered to the pillar that 20 2.5*50cm are housed, the simulated moving bed chromatography system that the ion exchange resin of having filled plumbous type sun is sorbent material, carries out chromatographic separation.65 ℃ of separation temperatures, feeding rate 150mL/h, pure water elution speed 1.0L/h.After system balancing, collect wood sugar and L-arabinose two-phase liquid, the purity of two-phase is all more than 95%, and it is more than 98% replying yield.

The L-arabinose of gained being adopted at 80 ℃ MVR machinery recompression high-efficiency evaporating system be concentrated into L-arabinose concentration is 800g/L, then according to the rate of temperature fall crystallisation by cooling of 3 ℃/h, when temperature is down to 50 ℃, add 0.1% crystal seed, crystal seed is educated after brilliant 2h, then the rate of temperature fall according to 2 ℃/h continues crystallisation by cooling, be cooled to 30 ℃ of left and right, by centrifugal after the L-arabinose crystal taking-up of separating out, the isolated L-arabinose of supernatant liquor and chromatographic system is merged mutually, repeat above-mentioned condensing crystal process, the centrifugal solid-phase obtaining rinses 2 times with dehydrated alcohol, and then 45 ℃ of drying in oven, L-arabinose crystallization yields is 65%, purity is more than 99%.

The wood sugar that xylose mother liquid separation is obtained returns mutually and is incorporated in the initial Xylose of the 4th step, adopting MVR machinery recompression high-efficiency evaporating system to be concentrated into xylose concentration is 800g/L, repeat xylose crystalline step, obtain wood sugar crystal and xylose crystallization mother liquor, xylose crystalline productive rate is 60%, purity is more than 99%, after the xylose mother liquid that the xylose mother liquid xylose crystalline initial with the 4th step that this step obtains obtains mixes, enter simulation moving-bed separation system and obtain wood sugar phase and L-arabinose phase, with this repeatedly.

Claims (1)

1. a production technique of preparing wood sugar and L-arabinose for clean and effective, is characterized in that, comprises the steps:

(1) agriculture and forestry organic waste material raw material carries out pre-treatment and dipping pretreatment; Described agriculture and forestry organic waste material raw material comprises: corn cob, maize straw, rice straw, jowar stalk, cotton seed hulls or bagasse;

Pre-treating technology comprises: the pulverizing of raw material, dedusting, washing three process;

The soak solution that described dipping pretreatment raw material is used is dilute sulphuric acid, dilute hydrochloric acid or rare oxalic acid, the massfraction of dilute sulphuric acid, dilute hydrochloric acid or rare oxalic acid is 0.1-0.5%, soak solution is with the massfraction of raw material than being 4-8:1, and the temperature of immersion is 30-100 ℃, and the time of immersion is 6-24h;

After dipping pretreatment finishes, solid-liquid separation obtains pretreated raw material, and the mode of isolated pretreated raw material by washing regulates pH value to 4.5-6.0, and soak solution is replied and utilized;

(2) the agriculture and forestry organic waste material raw material after process (1) takes the method hydrolyzed hemicellulose of high temperature steaming to obtain high temperature steaming Xylose or high temperature steaming xylo-oligosaccharide liquid, specific as follows;

Pretreated agriculture and forestry organic waste material is added in reaction unit, adds 5-20 doubly to the water of solid material dry weight; Initial pH is 5.0-6.0; Boiling 1h under the condition of 155 ℃; Mixing speed is 80-150rpm; Operating pressure is 0.8MPa, obtains xylo-oligosaccharide cooking liquor;

Or the agriculture and forestry organic waste material raw materials such as pretreated corn cob or cotton seed hulls are added in reaction unit, add 5-20 doubly to the water of solid material dry weight; At initial pH, be 4.5-6.0; High temperature steaming 2h under the condition of 165 ℃; Mixing speed is 80-150rpm; Operating pressure is 1.0MPa, obtains wood sugar cooking liquor;

(3) utilize milk of lime to above-mentioned high temperature steaming xylo-oligosaccharide liquid carried out neutralizing treatment to pH at 8.0-9.0, again with phosphoric acid or hydrochloric acid readjustment pH to 6.0, Plate Filtration obtains supernatant liquor afterwards;

Xylo-oligosaccharide cooking liquor after the crude enzyme liquid that employing aspergillus niger produces is processed (3) carries out enzymolysis and obtains wood sugar enzymolysis solution, specific as follows:

Cha Shi substratum, 115 ℃ of sterilizing 30min, access aspergillus niger is cultivated the aspergillus niger seed liquor that obtains having spore shape form for 3-4 days under the condition of 28-30 ℃; Product enzymic fermentation is cultivated: sucrose 2.4g, MgSO ₄0.12g, K ₂hPO ₄0.12g, KH ₂pO ₄0.24g, FeSO ₄0.0012g, NaNO ₃0.012g, uses 100ml water dissolution, mixes with the mixture of wheat bran with corn cob, and the mixture of corn cob and wheat bran amounts to 100g, and corn cob and wheat bran mass ratio are 3:7-7:3, and under the condition of 115 ℃, sterilizing 30min obtains the first fermention medium;

Under the condition of aseptic technique, in 100g the first fermention medium, add the aspergillus niger seed liquor 10-15ml with spore shape form, add sterilized water 30-90ml, take the mode of solid state fermentation, under the condition of 30 ℃, cultivate 3-4 days; Product enzymic fermentation under the condition of 28-30 ℃, adds 250ml water mechanical shaking extraction solid culture 1-2h after cultivating and finishing, and centrifugal collection supernatant liquor is crude enzyme liquid;

Crude enzyme liquid is joined in xylo-oligosaccharide cooking liquor, and enzymatic hydrolysis condition is temperature: 40-50 ℃, pH=5.0-5.5; Enzyme concentration 5-10ml in 100ml cooking liquor, the xylo-oligosaccharide concentration in cooking liquor is 1.5%, after enzymolysis 1-2 days, obtains Xylose;

Or utilize the above-mentioned high temperature steaming Xylose that calcium carbonate obtains (2) to carry out neutralizing treatment tune pH to 6.0-7.0, and Plate Filtration obtain supernatant liquor;

(4) add microorganism to remove the assorted sugar that comprises glucose and semi-lactosi in Xylose;

Selected microorganism is Angel high activity dried yeast; The required nitrogenous source that ferments is at least one in yeast extract paste, yeast powder, corn steep liquor, Dried Corn Steep Liquor Powder, soybean-cake flour, ammonium sulfate, urea;

Above-mentioned Xylose, through after being concentrated into total sugar concentration and being 100-200g/L, adds above-mentioned nitrogenous source fully to dissolve, and the nitrogen concentration obtaining is 10-20g/L, is mixed with the second fermention medium;

At glucose concn, be 20-30g/L, yeast extract paste concentration is to add Angel high activity dried yeast in the activation medium of 5-10g/L, the starting point concentration that makes Angel high activity dried yeast is 1-2g/L, in the shaking table of 30 ℃, rotating speed activation 2-4h with 200-300rpm, centrifugal 15min abandons supernatant liquor, recentrifuge 10min after distilled water wash for the cell of bottom, collecting cell mud;

In the second fermention medium, add cell mud 10-20g/L, leavening temperature 25-35 ℃, fermentation rotating speed 200rpm, air flow is 1-1.5vv ^-1min ^-1, fermenting, fermentation time is 6-12h;

(5) centrifugal after fermentation, then gac or polymeric adsorbent, or both are in conjunction with Xylose is carried out to purification process;

(6) after wood sugar purifying, carry out concentration; Xylose Content in the Xylose obtaining is 700-800g/L;

(7) take the mode of gradient programmed cooling crystallization to obtain xylose product and xylose crystallization mother liquor;

(8) xylose crystallization mother liquor obtaining is separated through chromatographic system, obtain wood sugar phase and L-arabinose phase;

(9) wood sugar step (8) being obtained is back to mutually step (6) and merges concentrated with wood sugar refined solution, take the mode of gradient programmed cooling to carry out crystallization, obtain wood sugar crystal and xylose mother liquid, xylose mother liquid in xylose mother liquid and step (7) merges, enter to chromatographic fractionation system separation, repeating step (8); The L-arabinose that (8) are obtained obtains L-arabinose product and L-arabinose mother liquor through concentrated, crystallization mutually, and L-arabinose mother liquor is replied and utilized.

Images