CN101544996A

CN101544996A - New process of plant material diastatic fermented koji (mycoprotein provender) and alcoholic fermentation production

Info

Publication number: CN101544996A
Application number: CN200810188381A
Authority: CN
Inventors: 李换位
Original assignee: Individual
Current assignee: Individual
Priority date: 2008-03-25
Filing date: 2008-12-26
Publication date: 2009-09-30

Abstract

A new process of plant material diastatic fermented koji (mycoprotein provender) and alcoholic fermentation production comprises the following steps: raw plant materials are immerged and wetted by 2.3-6.5 percent of sulphuric acid (or hydrochloric acid or nitric acid) water solution and then processed with ventilating air with the wetness fraction of 85-97 percent and the temperature of 35-65 DEG C under ultraviolet irradiation or processed with drying under a natural environment to obtain plant material preprocessing raw materials which are easy to pulverize; a diastatic fermented koji culture medium or an alcoholic fermented mash with a certain solid-to-liquid ratio is prepared by adding fixed-amount organic nitrogen materials or inorganic nitrogen materials into the pulverized raw materials, the pH value of the alcoholic fermented mash is neutralized by aqueous alkali, culture medium material inoculating trichoderma viride or aspergillus niger and microzyme are fermented together to prepare preprocessing plant material diastatic fermented koji under the temperature of 28 DEG C-35 DEG C, the alcoholic fermented mash is delivered to the diastatic fermented koji of the same and then fermented to prepare a ripe fermented mash under the temperature of 28 DEG C-35 DEG C, the ripe fermented mash is rectified to obtain azeotropic alcohol, and solid materials filtered from the ripe fermented mash are the plant material diastatic fermented koji or the mycoprotein provender after being dried under the temperature of 35 DEG C-45 DEG C.

Description

Plant materials diastatic fermentation distiller's yeast (tropina diet) and alcoholic fermentation new process of production

Technical field

The biomass material pre-treatment, the technologies such as alcoholic fermentation of CELLULASE and distiller's yeast (or tropina diet) production or biomass polysaccharide.

Background technology

Lignocellulose has constituted the cell walls of plant materials; pair cell plays provide protection; its main organism comprises Mierocrystalline cellulose, hemicellulose and xylogen three parts; hemicellulose and xylogen in the cell walls are formed network structure by covalent bond; Mierocrystalline cellulose is embedded in wherein; the content of Mierocrystalline cellulose, hemicellulose and xylogen in the different biomass is different; wherein hemicellulose is the mixture that is made of different saccharans; these saccharans are to be polymerized by different monose; there is straight chain that side chain is also arranged, is connected with the ethanoyl and the methyl of different quantities above.The hydrolysate of hemicellulose comprises that two kinds of five-carbon sugars (wood sugar and pectinose) and the various sugared proportions of three kinds of hexoses (glucose, semi-lactosi and seminose) change with raw material, general wood sugar accounts for over half, the pectinose (can account for the 10%-20% of five-carbon sugar) that also has a great deal of when being hydrolysis material with agricultural crop straw and grass, the polymerization degree of hemicellulose is lower, contained sugar unit number is at 60-200, there is not crystalline structure yet, so it is than facile hydrolysis, just can hydrolysis in diluted acid about 100 ℃, also can be in complete hydrolysis under the enzyme catalysis.And be joined together to form the cellulose bundle of crystalline structure between cellulose macromolecule by a large amount of hydrogen bonds.This structure makes cellulosic character very stable, water insoluble, and no reductibility is difficult for hydrolysis takes place at normal temperatures, and hydrolysis at high temperature is also very slow.Only having in the presence of the catalyzer, cellulosic hydrolysis reaction just carries out significantly.Catalyzer commonly used has mineral acid and cellulase.Acid hydrolysis process and enzyme hydrolysis process have been formed thus respectively.Xylogen be by the phenyl-propane structural unit by the C-C three-dimensional space macromolecular compound that is formed by connecting, but its molecular formula simple table is shown (C ₆H ₁₁O ₂) n, xylogen can not be hydrolyzed to monose, and forms protective layer around Mierocrystalline cellulose, influences cellulosic hydrolysis.Mierocrystalline cellulose belongs to macromolecular polysaccharide, is by gluconate dehydratase, the straight chain polymer that is formed by connecting by β 1,4 glucoside bond, but cellulosic molecule formula simple table is shown (C ₆H ₁₀O ₅) m, the m here is the polymerization degree, the number of glucose unit in the expression Mierocrystalline cellulose, and it is worth generally 3500-10000, and Mierocrystalline cellulose can generate glucose through hydrolysis.

The pre-treatment of plant materials raw material: pretreated purpose: pre-treatment that important topic is a raw material in the enzyme hydrolysis process, owing to constitute the Mierocrystalline cellulose of biomass major ingredient, twine mutually between hemicellulose and xylogen, and there is the product body structure in Mierocrystalline cellulose itself, can stop enzyme near cellulose surface, so efficient is very low during the biomass direct hydrolysis, can remove delignification by pre-treatment, dissolve hemicellulose or destroy cellulosic crystalline structure, can be thereby increase it near the surface, improve the hydrolysis productive rate, good pretreatment technology process should meet the following conditions: (1) can promote the generation of sugar or help the enzymic hydrolysis of back; (2) can avoid the degraded of carbohydrate to lose; (3) avoid generating to hydrolysis and the deleterious byproduct of fermentation; (4) reasonable economically.

Many pretreatment processs are studied, can roughly it be divided into physics method, physico-chemical processes, chemical method and biological process 4 classes.Because up to the present the pretreated speed of biological process is too slow, still in conceptual phase, so mainly be preceding 3 classes in the present industrial production.(1) physics method: the physics method mainly is a mechanical disintegration, can by cut, grind, technology such as mill diminishes the granularity of biomass material, increases and the surface in contact of enzyme, main is the cellulosic crystalline structure of destruction.Can make the granularity of raw material drop to 10-30mm by chopping, and by can reaching 0.2-2mm after milling, but it is bigger to pulverize the required energy consumption of biomass material.(2) physico-chemical processes: mainly comprise steam pop, ammonia burst, CO ₂Explosion etc.Steam explosion is with steam biomass material to be heated to 200-240 ℃, and keeps 0.5-20min.High temperature and high pressure causes the softening of xylogen, opens the valve step-down then rapidly, causes the explosion of Mierocrystalline cellulose crystalline, and xylogen is separated with Mierocrystalline cellulose, and the effect of water vapor explosion depends primarily on the granularity of the residence time, treatment temp, raw material and water ratio etc.Studies show that, at comparatively high temps with than short residence time(SRT) (270 ℃ 1min) handle down, or (190 ℃, 10min) treatment effect are all fine down at lesser temps with than long residence time.But there are some researches show recently, adopt lesser temps and more favourable than long residence time.The advantage of steam pop is that energy consumption is low, but intermittently also operate continuously.Mainly be suitable for hardwood raw material and agricultural crop straw, but to the effect of cork relatively poor (more because of content of lignin in the cork, also do not have the conduit of all length in the hardwood, so heat transfer efficiency is relatively poor, catalyzer transmits difficulty).Shortcoming is that the wood sugar loss is many, and produces the deleterious material that ferments.Pre-treatment intensity is big more, and cellulase hydrolysis is easy more, but the sugar that is obtained by hemicellulose is just few more, and the fermentation nuisance of generation is many more.

Ammonia burst principle is similar to steam pop, and it makes solid material and liquid ammonia react under high temperature and high pressure, open the valve step-down equally behind certain hour suddenly, causes the explosion of Mierocrystalline cellulose crystalline.Typical its treatment temp of technology is at 90-95 ℃, 20-30min holds time, every kilogram of solid material is with 1-2kg ammonia, for reducing the technology cost, ammonia needs to reclaim, and uses the overheated ammonia steam of temperature up to 200 ℃ with the ammonia recovery on the residual solid material, for making ammonia and water sepn for this reason, can get off most of water vapor condensation with precondenser earlier, stay purity up to 99.8% ammonia steam.Liquid in the precondenser enters rectifying tower, and cat head also can get 99.8% ammonia steam.These ammonia steams recycle after the condensation compression.The ammonia burst does not produce objectionable impurities, and the sugar loss in the hemicellulose is also few, but also undecomposed through the hemicellulose of this processing, needs use the hemicellulose enzymic hydrolysis in addition, so processing cost is bigger.CO ₂Explosion is similar substantially to the ammonia burst, just uses CO ₂Replaced ammonia, but its effect is than ammonia treatment difference.

Chemical method comprises alkaline purification, diluted acid processing and ozonize etc.Its alkaline purification method is to utilize xylogen can be dissolved in the characteristics of basic solution, handles biomass material with dilute sodium hydroxide or ammonia solution, destroys the wherein structure of xylogen, thereby is convenient to the carrying out of enzymic hydrolysis.The technology that comes into one's own is the method for handling with ammonia solution, and is volatile because of ammonia, can easily reclaim (rate of recovery is more than 99% in batch experiment) by heating, and pretreating effect is fine.As with ammonia treatment poplar raw material, 180 ℃ handle 1 hour down after, fiber wherein have 95.2% can be by enzymic hydrolysis, and 7.8% hydrolyzable only when being untreated.Can also reclaim the higher xylogen of purity as industrial chemicals by the ammonia pre-treatment.Dilute acid pretreatment is similar to acid hydrolysis, by being monose with the hydrolysis of hemicellulose in the raw material, reaches the purpose that makes raw material mix loose.The liquid glucose that hydrolysis the obtains preparing alcohol that also can ferment.The cork hydrolysis is difficulty, need under stronger condition, to carry out pre-treatment, at this moment also can adopt the method for two-stage dilute acid pretreatment, promptly the raw material of handling through first step washing earlier, under stronger condition, carry out the second time again and handle, can reduce the decomposition of sugar and the generation of nuisance like this.

Can remove delignification effectively with ozonize, reaction is carried out at normal temperatures and pressures, also can not produce objectionable impurities, but cost is too high, impracticable.

Cellulase production: most of bacterium can not be decomposed the Mierocrystalline cellulose of crystalline structure, and because of their enzyme system imperfection, but some mould can secrete the required whole enzymes of hydrocellulose, and these enzymes are very important to cellulase production.At present the maximum of usefulness are Trichodermareesei (T.reesei).Various Institute of Micro-biology excretory cellulase is incomplete same.Highly active inscribe glucolase and circumscribed glucolase can have been produced as many Li's Trichoderma strains, but the β glucuroide that they produced is active relatively poor, though and belong to the cellulolytic ability of mould of Aspergillus system, but the ability of decomposing cellobiose is very strong, just can be the cultivation of putting together of these two kinds of bacterial strains when the production of cellulose enzyme.The production of cellulase is solid state fermentation and two kinds of methods of liquid state fermentation.(1) solid state fermentation: so-called solid state fermentation is meant that microorganism grows on the matrix of free-water not having, the similar wheat bran production of this process, its advantage is that energy consumption is low, low to ingredient requirement, enzyme concn height in the product can be directly used in hydrolysis, and shortcoming is required artificial many, be difficult for polluting control, each batch product property poor repeatability.

(2) liquid state fermentation: the scale operation cellulase is based on the liquid submerged fermentation method at present, and the advantage of liquid state fermentation is required artificial few, is suitable for scale operation, easily pollutes control.Each batch product property good reproducibility.Shortcoming is that energy consumption is big, the ingredient requirement height, and enzyme concn is low in the product.No matter be the sort of fermentation process, general all using through pretreated biomass is the matrix of production enzyme, this is except can reducing cost, the benefit that also has other two aspect: (1) microorganism can be a carbon source with the pretreated biomass of process well, the enzyme produced thus more is applicable to cellulose hydrolysis than producing enzyme with pure cellulose or sugar during by matrix, and activity is higher.(2) more convenient on flow arrangement with same raw material production enzyme and production alcohol, can reduce equipment cost.The microorganism growth that also has that also will add fermentor tank is simultaneously bred required mineral nutrition thing and tensio-active agent etc.But the production cost of cellulase is still too high at present.

Cellulase hydrolysis: in the enzymic hydrolysis process of cellulose substances, it is generally acknowledged that enzymic hydrolysis comprises three primary processes: the absorption of enzyme on solid material, enzymatic hydrolysis and enzyme desorption.Cellulosic enzymic hydrolysis is different with general enzymatic reaction, its substrate material Mierocrystalline cellulose is insoluble, enzyme just can work after only being adsorbed on its surface, so the absorption of enzyme is very big to the enzymic hydrolysis effectiveness affects, enzyme on being adsorbed on Mierocrystalline cellulose, also can be adsorbed on the xylogen, this is the important factor that influences enzymic hydrolysis efficient.It is found that hydrolysis rate was very fast when content of lignin was low in the raw material,, add the irreversible adsorption raising hydrolysis efficiency that tensio-active agent also can reduce enzyme so it is favourable to hydrolysis to remove delignification in pre-treatment.Add the better effects if of nonionic agent.In addition, hydrolysate has inhibition to the activity of cellulase, and the effect of cellulase also is subjected to the influence of other unwanted component, as strong oxidizer, reductive agent, metal ion etc., the enzyme that is combined on the Mierocrystalline cellulose is not subjected to this influence, and cellulosic speed of enzyme hydrolysis is also relevant with conditions such as cellulose concentration, enzymic activity, hydrolysis temperature and pH values.So in the ordinary course of things, make enzyme and Mierocrystalline cellulose thorough mixing, cellulose concentration is at most at 6%-10% (W/V) in the hydrolysis, and the consumption of cellulase is used filter paper enzyme activity representation unit FPU usually.The ability of its expression enzymic hydrolysis filter paper (cellulosic material that a kind of polymerization degree is lower) can be measured by experiment, and obviously enzyme dosage is many more, and hydrolysis rate is fast more.But the also corresponding increase of cost.In general experiment with when doing economic analysis, often the consumption of enzyme is decided to be 10FPU/ gram Mierocrystalline cellulose.Because of can having made Mierocrystalline cellulose, such consumption in 48-72 hours, reaches maximum transformation efficiency.The optimal temperature of enzymic hydrolysis is at 45-50 ℃, and required best pH value is about 4.8.

Alcoholic fermentation: the fermentation in the biomass waste preparing alcohol technology and be that the fermentation of raw material has a great difference with starch or sugar.This shows that mainly following two select: 1, regular meeting is contained the deleterious component of organism of fermentation in the biomass by hydrolyzation liquid glucose; 2, contain more wood sugar in the hydrolysis sugar liquid.Above characteristics have determined nuisance in this technology to remove the importance with wood-sugar fermentation, although the research wood-sugar fermentation is also arranged, general what consider all is the common fermentation of glucose and wood sugar.If only use glucose fermentation, generally all use yeast.Because of it has good alcohol conversion performance, be the most frequently used.Fermentation itself is oxygen consumption not, but the microbial growth breeding needs the certain oxygen of consumption, keeps a certain amount of oxyty in the fermented liquid so need make.(the biomass energy modernization utilizes technology/Wu to create it, Ma Longlong edits----Beijing 2003,5 biomass fuel alcohol 173-186 of Chemical Industry Press)

Whether suitable the diastatic fermentation condition of starchy material: whether diastatic fermentation is normal, will see the condition biological enzyme of creation and the requirement of microorganism.Generally at 20-70 ℃, optimum temperuture is 55-60 ℃ to saccharifying enzyme to the operative temperature scope of starch, and it is very fast to surpass 60 ℃ of enzyme activity inactivations, is lower than 20 ℃ of saccharification speed then relatively slowly, and it is longer thoroughly starch is converted into the required saccharification time of glucose.Saccharomycetic growth metabolism temperature range is generally at 15-41 ℃, optimum temperuture is 28-30 ℃, it is dead easily to surpass 41 ℃ of yeast, the breeding metabolic capacity obviously descends, being lower than 15 ℃, then to breed accretion rate slower, is unfavorable for that fermentation produces wine, and they are relatively more responsive to temperature, require strictness, and pH value can adapt to its requirement in the 4.0-6.5 scope.Therefore, the diastatic fermentation condition generally is regulated to suitable with the zymic optimum temperuture, promptly controls room temperature 25-35 ℃, and 28-35 ℃ of diastatic fermentation wine with dregs product temperature, charge amount are about 80% of fermenting container volume fraction.

The manufacturing of yeast saccharomyces cerevisiae: replace Ka Shi jar seed to make the vat distiller's yeast with the keen-witted and capable yeast of active wine

(1) active dry yeast activation: the active dry yeast that takes by weighing converted mash (the saccharification liquid of 1kg starchy material+5kg water) raw materials quality 0.1% adds 10 times of water and 10 times of converted mashs, joins in the converted mash standby at the 28-30 ℃ of following 8-10h of cultivation then about 35 ℃ behind the activation 3min.

(2) the solid wheat bran is cultivated: wheat bran is added water 75%-80% turn evenly, require no sandwich no agglomerate, under normal pressure, steamed material 1-1.5 hour then, steam about material postcooling to 30 ℃, insert the head mold song of activation culture dry yeast liquid 5%-10% and 0.1%-0.2%, fully mix thoroughly, require loose no agglomerate, material is poured in the ventilation pond left standstill cultivation then, culture temperature is controlled at 28-30 ℃, after the product temperature rose in 6-8 hour, begin to ventilate and suitably turn, turned and ventilated in 10-12 hour, the vigorous breeding of yeast cell about 15 hours, the product temperature rises very fast, will lower the temperature this moment to ventilate and turn, and yeast culture finished in 24-30 hour.

(3) oven dry: oven dry is general to divide two stages, begin to baking bent about 24 hour from advancing drying room early stage, at this moment water content is many in bent, still have part nutriment to be utilized in the matrix, bacterial classification continues the breeding growth, but excess moisture in bent, also easily cause assorted bacterium breeding, so early stage, bake out temperature should be grasped at 35-40 ℃, promptly played the cultivation effect, can reduce bent interior moisture again as early as possible, straight oven dry was the later stage baking stage from 24 hours, in this period, bent interior moisture is less, and thalline stops growing gradually, because moisture reduces, somatic cells strengthens the resistivity of heat, and bake out temperature at this moment generally is controlled at 40-45 ℃, and song is parched, to song moisture below 10%, just can go out song, also can utilize sunlight to dry, reach drying purpose.(brewing wine with raw materials technology/Huangping chief editor---Beijing: China Light Industry Press, calendar year 2001, koji for uncooked material production technique 115-168)

Cellulose solid-state fermentation: the cellulose solid-state fermentation microorganism is the fungi of Trichoderma, and the cellulose solid-state fermentation carbon source is added a small amount of wheat bran generally based on cellulosic material.With straw, weeds is carbon source, is nitrogenous source with ammonium sulfate, cultivates 96 hours for 30 ℃, and enzyme activity is all higher, adds wheat bran in the weeds, and thalli growth is rapid, and enzyme activity is the highest, and comparatively ideal ammonium sulfate consumption is for cultivating the 0.8%-1.0% of base unit weight.Cellulose solid-state fermentation technology controlling and process: in the suitability for industrialized production of reality, owing to be difficult to reach the product temperature control level that similar tray is cultivated, therefore the product temperature is controlled at 28-33 ℃ at earlier fermentation (24 hours), ferment middle (24-60 hour) is controlled at below 40 ℃, and strategy and operation that fermentation later stage (60-96 hour) is controlled at below 35 ℃ are acceptables.Its fermentation initial p H value is at 3.5-6.5, and the control of solid state fermentation product temperature mainly is by ventilating, turn over song and realize that generally ventilate for the first time in fermentation beginning in about 24 hours, each ventilation time is no more than 3min, turn over the song time ferment 40-48 hour better.Generally between 3.5-4.5, the fineness of fermentation materials generally is to get final product by the 35-45 mesh sieve to pH value in actual production process.The water content of fermentation materials is at 50%-60%.

Aspergillus niger also produces enzymes such as dextranase, cellulase, mannase, polygalacturonase and the amylase system of high enzyme vigor except producing aspartic protease and zytase in addition, produces complex enzyme for feed so Aspergillus is fit to solid fermentation very much.

Cellulase production technology has two kinds both at home and abroad at present: solid state fermentation and deep layer liquid state fermentation.Solid state fermentation production of cellulose enzyme has distinct advantages: 1, the solid state fermentation conditions environment more approaches the wooden mould habit under the state of nature, makes the enzyme system of its generation more complete, helps the natural cellulose of degrading; 2, the solid state fermentation consumes energy is few, and facility investment reduces relatively; 3, enzyme product yield height, the subsequent extracted process is handled well than liquid state fermentation.Therefore producing cellulase through solid-state fermentation more and more comes into one's own.The microorganism of occurring in nature biodegradable fiber element is a lot, comprises bacterium, fungi and actinomycetes, and cellulosic microbial destruction has two kinds of possible modes, and a kind of is to destroy the outside, then to in-house development; Another kind is to corrode from inside to outside.Mould is the microorganism of degraded cellulose, and it is when the substrate of degraded cellulose, and mycelia crosses secondary wall and enters cell, and constantly growth, and degraded cellulose makes Mierocrystalline cellulose progressively destroyed from inside to outside.A common characteristic is arranged during the microbiological degradation Mierocrystalline cellulose, the plain enzyme of promptly synthetic born of the same parents' outer fiber, but a spot of microorganism synthetic cellulose enzyme bunch degraded crystalline cellulose is arranged.(solid-state fermentation technology and application/Wu Zhenqiang chief editor-Beijing: Chemical Industry Press in February, 2006, the solid state fermentation 173-197 of zymin)

In order to reduce the production cost of alcohol, developed the zymotechnique of synchronous glycosylation (SSF) in the seventies in 20th century, promptly pretreated biomass, cellulase and organism of fermentation are added in the fermentor tank, enzymic hydrolysis and fermentation are finished in same device, and the also available several fermentor tank series connection of certain actual SSF flow process are produced.This technology has not only been simplified production equipment, and because of the cellulose hydrolysis speed in the fermentor tank far below glucose fermentation speed, make in the solution concentration of glucose and cellobiose very low, this has just eliminated them as the restraining effect of hydrolysate to enzymic hydrolysis, the corresponding consumption that reduces enzyme.Low in addition glucose concn has also reduced the chance of microbial contamination.SSF has become up-and-coming biomass alcohol technology at present.

The subject matter of SSF technology is the coupling of hydrolysis and fermentation condition.The required best pH value of cellulase hydrolysis is about 4.8, and the best pH value of fermentation is between 4-5, the two and noncontradictory, but the optimum temps of enzymic hydrolysis is at 45-50 ℃, and the optimum temps of fermentation is at 28-30 ℃, the two can not mate.Actual SSF is everlasting 35-38 ℃ and operates down, and this compromise treating processes makes the activity of enzyme and the efficient of fermentation all can not reach maximum.

In general SSF technology, the liquid that is rich in five-carbon sugar that pre-treatment produced is fermentation separately.Exploitation along with the novel microorganism of while glucose fermentation and wood sugar has developed SSCF technology again, and in this technology, liquid glucose that pre-treatment obtains and the Mierocrystalline cellulose of handling are placed in the same reactor to be handled, and this has just further simplified flow process.The raw materials pretreatment of this technology is the biomass 0.5%H after crushed ₂SO ₄Handle 10min down at 190 ℃, adopt the cocurrent flow type hydrolysis reactor, its form class is like the pulp digester in the paper industry.Wherein solid concentration is 20%, and this process can be 75% hydrolysis of hemicellulose a monose when destroying the Mierocrystalline cellulose crystalline structure, but also has 10% wood sugar and pectinose to be converted into furfural, has 15% semi-lactosi and seminose to be converted into HMF.The raw material that goes out pretreater enters flasher, and pressure drops to normal pressure from 1.2Mpa.This process also can remove a large amount of furfurals that produce in the pre-treatment, HMF and part acetate except lowering the temperature, and this will help follow-up workshop section.Enter strainer then and carry out liquid-solid separation, small portion in the gained pulpous state solid product is used to make cellulase, major part enters the SSCF reactor, the liquid spent ion exchange resin that filtration obtains is handled, remove 88% acetate and whole sulfuric acid, earlier this liquid pH value is adjusted to 2 then with sulfuric acid, with lime pH value is adjusted to 10 again, can remove the fermentation objectionable impurities in the liquid with calcium sulfate by this excessive alkali adding method, then the pH value of liquid is adjusted to 4.5, also it is joined the SSCF reactor.

Use 3 groups of 3600m in the SSCF workshop section ³Stirring-type stainless steel fermentation container, 6 of each groups, totally 18 fermentation containers.Enzyme dosage is a 15FPU/ gram Mierocrystalline cellulose.Fermentation is transgenosis Z.moblis with bacterial classification, adopts 2 groups of seed culture devices, and every group comprises 5 containers, enlarged culturing step by step, and every grade of inoculum size is equivalent to 10% of container volume.The SSCF operational condition is: 30 ℃ of temperature, and solid starting point concentration (comprising soluble and insoluble) 20%, the theoretical yield that residence time 7d, glucose and wood sugar are converted into alcohol is respectively 92% and 85%.

The enzyme production of SSCF is with 11 1000m ³The inflatable fermentation container, adopt periodical operation, there have 8 fermentation containers to be at any time to be in service, is bacterial classification with T.reesei (Richter scale wood enzyme).Adopt 3 groups of seed culture devices, every group comprises 3 containers, enlarged culturing step by step, and every grade of inoculum size is equivalent to 5% of container volume.The plain concentration of fermentation container initial fiber is 4%, and average every gram Mierocrystalline cellulose and hemicellulose can be produced the 200FPU cellulase, and fermentation productivity is 75FPU/Lh.

Fermented liquid makes azeotropic alcohol with traditional rectificating method, makes 99.9% alcohol with molecular sieve dehydration again, and the gasoline of adding 5% is made methylated spirits and stored.Waste liquid is through centrifugation and evaporation at the bottom of the rectifying tower still, and recyclable 80% water cycle is used.When bringing up to 45%, the solids concn in the lignin residue sends to combustion power generation.

At present still underway to the improvement of Production of Fuel Ethanol technology, a possible technology is associating biological treatment (CBP), also claim direct microbe transformation method (DMC), it is combined in cellulase production, cellulose hydrolysis, glucose fermentation and wood-sugar fermentation in the reactor and finishes, and only uses a kind of microorganism.This Process Plans hydrolysis productive rate reaches 92%, and fermentation production rate reaches 90%, and ethanol concn reaches 5%, and fermentation time is 36h, and employing is continuously fermented, and need not plant female jar.DMC has certain foundation, known have many microorganism handlebar cellulosic materials to be converted into the ability of alcohol, as mould Fusariumoxysporum, and bacterium Clostridia sp etc., but the ethanol production of existing DMC bacterial classification is all lower, and can not handle biomass material under low liquid-solid ratio.(the biomass energy modernization utilize technology/Wu create it, Ma Longlong chief editor-Beijing: in May, the 2003 biomass fuel alcohol 189-195 of biological chemistry press)

Single cell protein solid state fermentation production technique:

The maize straw solid state fermentation is produced SCP:

This technology is divided five parts: raw materials pretreatment, bacterial classification enlarged culturing, solid-state ventilating fermentation, two bacterial strain step by step ferment and the product aftertreatment altogether.1, raw materials pretreatment: maize straw is pulverized after the high compressed steam explosion treatment is equipped with auxiliary material, water-wet, mixes the back vapor sterilization thoroughly.2, enlarged culturing: A and B two bacterial strains carry out triangular flask, lunch box kind song, koji tray kind song enlarged culturing step by step by separately culture condition respectively to bacterial classification step by step.3, solid-state ventilating fermentation: adopt rotary disc starter propagation machine, inoculation cellulase high-yield 96-32 carries out the solid-state ventilating fermentation of thick-layer, to obtain to contain the prozyme song of greater activity cellulase, amylase, proteolytic enzyme etc., and the enzymolysis cellulosic material, produce materials such as monose and amino acid.4, two bacterial strains ferment altogether: with A bacterial strain solid state fermentation product is matrix, and inoculation yeast bacterium B carries out two bacterial strains and ferments altogether, enzymolysis and bacterial strain propagation is carried out simultaneously, to obtain high enzyme living single-cell protein product.5, product aftertreatment: tunning promptly gets product through cryodrying, pulverizing, batch mixes altogether.(solid-state fermentation technology and application/Wu Zhenqiang chief editor-Beijing: Chemical Industry Press in February, 2006, the solid state fermentation 254 of feedstuff protein)

From above-mentioned summary, clear and definite recognize existing the unfavorable factor that overcomes and avoid of being difficult in the above-mentioned biomass saccharification alcoholic fermentation technology, and its technological process is numerous and diverse, though as the biomass preprocessing process is that hemicellulose obtains complete hydrolysis, but the loss that causes plant materials hemicellulose polysaccharide is up to 25%, and make its material that has generated harmful fermentation, and must remove processing.This can increase facility investment and the technological process energy consumption and the process cost of its technology undoubtedly.And in its zymotechnique, the sort of microorganism that don't work carries out alcoholic fermentation, and the suitableeest leavening temperature of its organism of fermentation and the suitableeest hydrolysis temperature of cellulase saccharification all can not mate, and have to adopt the method controlled temperature of compromise.And the diastatic fermentation temperature of this compromise makes the activity of its saccharifying enzyme and fermentation efficiency all can not reach maximum, and influences the saccharification and the fermentation efficiency of its technology.Have a weakness and its DMC is same, that is exactly that the ethanol production of DMC bacterial classification is lower, and can not handle biomass material under low liquid-solid ratio.And just at present many unfavorable factors of biomass saccharification fermentation alcohol production technique restricting the popularity of biomass fuel alcohol industry development.So, in order to be converted into repeat regenerated large number of biological raw material year after year human inexhaustible, nexhaustible biomass fuel alcohol and diet raw material, its plant saccharan is converted into is come high-effect production fuel alcohol or albumen diet by the fermentable sugars that the ethyl alcoh(ol) yeast fermentation utilizes, this need develop a kind of pretreatment technology of brand-new biomass material, with its realizing this goal with common saccharomycetes to make fermentation with its brand-new diastatic fermentation distiller's yeast or tropina diet production technique, Here it is aim of the present invention and purpose are so the present invention has produced.

Summary of the invention

Plant materials raw materials pretreatment processing: through dust separating tearing (standing grain this) or cut into slices and grind (woody) plant materials raw material, concentration be in 2.3%-6.5% mineral acid (perhaps hydrochloric acid or sulfuric acid or the nitric acid) aqueous solution behind even dipping, being placed on temperature is 35 ℃--65 ℃, humidity is handled under uviolizing in the ventilating air of 85%--97%, certain hour, perhaps be placed on temperature more than 25 ℃ in the spontaneous current blowing air, through diurnal cycle airing certain hour, after all being easy to pulverize to the plant materials raw material, carrying out drying and grind.Through the degraded of its hemicellulose glycocalix of this pretreated plant materials, and its five-carbon sugar molecular structure changes, and the sugared body that this molecule isomery changes can be utilized by the metabolism of ethyl alcoh(ol) yeast fermentation.Change has also taken place in the structure of its cellulose macromolecule, the change of this molecular structure make its polysaccharide body can be under 35 ℃ be institute's hydrolysis by Trichoderma and aspergillus niger microbial compound enzyme, and its lignin structure has suffered that also disassociation destroys.

The diastatic fermentation distiller's yeast tropina diet of pre-treatment plant materials) produce: in the plant materials material through adding inorganic nitrogen material or a certain amount of organonitrogen material (should comprise microorganism culturing grow required somatomedin), neutralize its pH value to 4-5 with alkali aqueous solution after adding its massfraction 70%-80% water again, insert solid masses 0.3%-0.5% then through three grades progressively behind the kind song of enlarged culturing (Trichoderma and aspergillus niger ratio are 1: 1 composite bacteria song), insert in the ventilating fermentation pond and cultivate, in earlier stage (24h) control product temperature is at 28-30 ℃, mid-term, (24-60h) controlled 30-35 ℃ of product temperature, later stage, (60-72h) control product temperature were at 30-33 ℃, and beginning ventilation for the first time behind 24h simultaneously, each ventilation time is no more than 3min, turning over the bent time carries out after cultivating 40-42h, insert again when turning through enlarged culturing distillery yeast liquid 4%-8% step by step, entering many bacterial classifications ferments altogether, fermentation 30-32h, turning over bent number of times during this time is advisable at 3-4 times, the water content of fermentation materials is controlled at 50%-60%, dry after the fermentation ends, oven dry divides two stages, begin to baking bent 24h from advancing drying room early stage, this stage bake out temperature is controlled at 35 ℃-40 ℃, behind 24h, be the later stage up to oven dry, this phase bake out temperature is controlled at 40 ℃-45 ℃, song is parched to song moisture content just can go out song 10%-15%, also can utilize sunlight to dry, reach drying purpose, pulverize after the song oven dry, product Wen Buying is above 55 ℃ during pulverizing.This pulverizes bent material and is the diastatic fermentation distiller's yeast of pre-treatment plant materials or tropina diet.

The alcoholic fermentation technology of pre-treatment plant materials raw material: after adding a certain amount of organonitrogen material (should comprise microorganism culturing grow required somatomedin) or inorganic nitrogen material in the pre-treatment plant materials material, add 2.2-4 times of water of its quality, with alkali aqueous solution its pH value that neutralizes is 4-5, and the diastatic fermentation distiller's yeast that inserts its solid material massfraction 0.6%-1.2% then carries out diastatic fermentation.The primary fermentation phase, the product temperature control was at 28-30 ℃, this yeast phase is 24h, the temperature control of lord ferment period product is at 30-35 ℃, should stir mash 1-2 every day during this, and keep 2-3d, the secondary fermentation phase, the product temperature control was at 30-33 ℃, time need be kept 1-2d fermentation ends, fermentation liquid distills after sterile filtration can get azeotropic alcohol, make 99.9% spirituous solution again with molecular sieve dehydration, this leaches solids and promptly obtain pre-treatment plant materials diastatic fermentation distiller's yeast or tropina diet after 35 ℃ of-45 ℃ of oven dry.But this technology also semi continuous operation is handled and multistage full operate continuously processing, the technological process that its semi continuous operation is handled is: its single-stage periodical operation treating processes is finished the resulting ripe mash in back not exclusively send to distillation, use as the new diastatic fermentation bacterial classification song that adds pre-treatment plant materials material and carry out the repetition periodical operation but stay sub-fraction.So just save the material desired microorganisms that ferments again, and can save a large amount of diastatic fermentation distiller's yeast raw materials, realized reducing the purpose of technology cost.Its continuous processing manipulation is that the prior fermentation mash that is obtained in the treating processes its intermittence is arrived the next stage reactor through overflow (or conveying) on the basis of adding virgin material continuously, carry out the fermentation in mid-term again, Deng finish the fermentation in mid-term to the mash of second stage reactor after, overflow again (or conveying) is in the next stage reactor, carry out later stage fermentation, after finishing third stage fermentation, getting final product overflow (or carry) has been filtering workshop section and has carried out rectification process and processed, so just realize the continuity of this saccharification and fermentation, and operation preferably control.But need the pollution of the assorted bacterium of strict control, require manipulation under the height sterile environment.Certainly this complete continuous treating processes also can be after its each yeast phase be all finished, the adding mode of its plant materials diastatic fermentation distiller's yeast, and available the backflow with a certain amount of fermentation liquid equivalence of diastatic fermentation distiller's yeast realized.

The technological effect of the alcoholic fermentation technological process of the diastatic fermentation distiller's yeast from the pretreatment technology process of above plant materials raw material of the present invention to pre-treatment plant materials raw material or the technological process of pretreating raw material plant materials tropina diet and pre-treatment plant materials, technology of the present invention is compared with present each existing technology has following superior part: plant materials pretreatment technology process of the present invention is simple, technological equipment investment is low, technological process is gentle and do not produce the material of harmful fermentation, there is not the loss of sugar, and on the basis of hemicellulose degraded, its wood sugar and pectinose isomerization and can be utilized by the metabolism of ethyl alcoh(ol) yeast fermentation, and its cellulose polysaccharide macromolecular structure also changes.This change makes it reduce the suitable hydrolysis temperature of cellulose complex enzyme as enzyme substrates, culture temperature when making cellulase production and cellulase hydrolysis temperature and diastatic fermentation temperature can be mated mutually, thereby have improved the production efficiency of its fermenting process.And making not anti-nutrition sugar body existence in the diastatic fermentation distiller's yeast of making (or tropina diet), this all is existing inaccessible effects of each technology.

And in the saccharification and fermentation process process of the present invention, its diastatic fermentation bacterial classification is to add with the form of song, thus obviously its enzymic hydrolysis efficient than directly adding high many of its hydrolysis efficiency of cellulase.This is similar to existing DMC technology, but than big many of its solid-to-liquid ratio of DMC technological process, also high many of ethanol production.Thereby its diastatic fermentation efficient is also all than each existing technology height.The solid-to-liquid ratio of existing technology is 1:5, and solid-to-liquid ratio of the present invention is about 2:5.So diastatic fermentation preparing alcohol technology of the present invention can make its rectification process cost reduce more than 50% than existing technology.Fermentation efficiency improves more than 50%, and sugared utilization ratio can directly improve 25%, and its continuous conversion zymotechnique cost is lower.

And the diastatic fermentation distiller's yeast that it is made is compared sugared utilization ratio height as the tropina fodder product with existing technology.Cost is reduced, and product do not contain anti-nutrition sugar body, as long as add competent calcium constituent and an amount of dl-methionine and vitamins B ₁₂, just can obtain the albumen fodder product with the same nutrition of fish meal, but its value is high more a lot of than fish meal.And the present invention makes pre-treatment plant materials diastatic fermentation distiller's yeast or tropina diet and alcoholic fermentation be produced in one, and this is the significant effect of the present invention.

Embodiment

Get the 1000 gram dedusting gramineae plant bodies that prescind or tear up or 1000 gram wood sawdusts (or wood shavings) respectively after soaking into 2.3%-6.5% sulfuric acid (perhaps hydrochloric acid or the nitric acid) aqueous solution, insert in the baking oven respectively, keep humidity in the baking oven 85%-97% simultaneously, make it under the UV-lamp irradiation, handle after all being easy to inside and outside the plant materials to pulverize with 35-65 ℃ ventilating air, drying grinds, or under physical environment temperature more than 25 ℃, shine system respectively and repeat airing through the ageing of spending the night, after same airing all was easy to pulverize to the plant materials inside and outside, drying ground.

Make the diastatic fermentation distiller's yeast or the tropina diet of pre-treatment plant materials material: get 500 gram pre-treatment and grind plant materials, standing grain basis and woody 8%-28% or the 35%-48% wheat bran of adding respectively, the molconcentration that adds behind the 70%-80% water with ammonium and calcium neutralizes its pH value to 4-5 than the alkali aqueous solution at 1:7, after inserting that solid masses 0.3%-0.5% wood is mould and being the kind song of 1:1, insert in the ventilating fermentation case and cultivate with the aspergillus niger ratio.In earlier stage (24h) control product temperature is at 28-30 ℃, mid-term, (24-60h) controlled 30-35 ℃ of product temperature, later stage (30-33 ℃ of 60-72h) control product temperature, and beginning ventilation for the first time behind 24h simultaneously, each ventilation time is 3min, turns over the bent time and carries out after cultivating 40-42h, inserts through enlarged culturing distillery yeast liquid 4%-8% step by step when turning again and the water content of fermentation materials is controlled at 50%-60%, turn over bent number of times 3-4, many bacterial classifications 30-32h that ferments altogether finishes.Go out bent oven dry then, drying baker is put in song after cultivating carried out drying treatment, in earlier stage (24h) interior bake out temperature is at 35-40 ℃, then bake out temperature is transferred to 40-45 ℃ and parches to song, and make the water content of song just can go out song at 10%-15%.Pulverize after going out song, the temperature of Qu Pin is 55 ℃ during pulverizing.Song after drying grinds is the diastatic fermentation distiller's yeast or the tropina diet of pre-treatment plant materials material.

Pre-treatment plant materials material alcoholic fermentation: get pre-treatment plant materials material 500 grams, add 8%-28% wheat bran (standing grain originally) or 35%-48% wheat bran (woody), then this material is joined in 30-35 ℃ the water of 2.2-4 times of its quality, neutralize its pH value to 4-5 with ammonium hydroxide and calcium hydroxide molconcentration than the alkali aqueous solution of 1:7 then, through adding its solid masses 0.6%-1.2% diastatic fermentation distiller's yeast, make diastatic fermentation distiller's yeast or tropina diet and alcoholic fermentation and produce mash, insert after mash is mixed thoroughly in the fermentation container and (replace) with 3000 milliliters there-necked flasks, behind 28 ℃ of-30 ℃ of bottom fermentation 24h, the product temperature is transferred to 30-35 ℃ of fermentation 2-3d, this yeast phase should stir 1-2 times every day, and then the product temperature is transferred to 30-33 ℃ of fermentation 1-2d secondary fermentation finishes.This fermenting-ripening mash obtains azeotropic alcohol through sterile filtration rectifying, and this alcohol can obtain 99.9% spirituous solution again after dehydration.The solid substances that filters acquisition after 35 ℃ of-45 ℃ of dryings, is promptly obtained pre-treatment plant materials diastatic fermentation distiller's yeast or tropina fodder product.

This is the single-stage periodical operation process of this technology, certainly this technology also can be carried out processing of single-stage semi continuous operation and multistage full operate continuously processing, the technological process that its semi continuous operation is handled is: above-mentioned single-stage periodical operation treating processes is finished the resultant ripe mash part in back send to rectifying, partly adding virgin material again forms new fermentation liquid in addition, carry out single stage operation again, the same semicontinuous fermentation that carries out of control process.Its continuous processing manipulation is that the prior fermentation mash that is obtained in the treating processes its intermittence is arrived next stage reactor (there-necked flask) through overflow (or conveying) on the basis of adding virgin material continuously, carry out the fermentation in mid-term again, Deng finish the fermentation in mid-term to the mash of second stage reactor (there-necked flask) after, overflow again (or conveying) is carried out later stage fermentation in the next stage reactor, after finishing third stage fermentation, get final product overflow (or carry) and be filtering workshop section and carry out rectification process and processed, the red-tape operati temperature and time is the same.This crosses range request manipulation under the height sterile environment.Certainly this complete continuous access of its diastatic fermentation distiller's yeast for the treatment of processes continuously, be its finish-reflux by a certain amount of fermentation liquid of equivalence behind the full phase fermenting process and finish.

The use of above technological process organic nitrogen source material, other organic nitrogen source material compound of also available and the equivalence of wheat bran amount nutrition balance.

Claims

1, plant materials diastatic fermentation distiller's yeast (tropina diet) and alcoholic fermentation new process of production are tearing up or cut into slices and grind the plant materials raw material through dust separating, soaking into wetting back with 2.3%-6.5% sulfuric acid (perhaps hydrochloric acid or the nitric acid) aqueous solution is 85%-97% in humidity, temperature is to handle continuously under 35 ℃-65 ℃ ventilating air middle-ultraviolet lamp shines or solarization system naturally under temperature environment more than 25 ℃, and repeat airing through the ageing of spending the night and handle, after all being easy to inside and outside the plant materials to pulverize, drying grinds, this ground material is plant materials diastatic fermentation distiller's yeast or tropina diet production pre-treatment base-material, also produces pretreating raw material for the plant materials alcoholic fermentation.This base-material and raw material are through adding a certain amount of organonitrogen material (should comprise microorganism culturing grow required somatomedin) inorganic nitrogen material, water, transfer its pH value to 4-5, access composite bacteria song, just can carry out diastatic fermentation distiller's yeast (or tropina diet) produces or alcoholic fermentation production, it is characterized in that: will add 8%-28% wheat bran (standing grain originally) or 35%-48% wheat bran (woody) in the pre-treatment plant materials material, neutralize its pH value to 4-5 with ammonium hydroxide and the alkali aqueous solution of calcium hydroxide molconcentration after adding 70%-80% water again than 1: 7, and after stirring, then, with evenly insert in this material its solids content massfraction 0.3%-0.5% through multistage progressively enlarged culturing kind song (Trichoderma and aspergillus niger ratio are 1: 1 composite bacteria), cultivate at 28 ℃ of-35 ℃ of bottom fermentations, inoculate behind fermentation culture 40-42h and carry out many bacterial strains through enlarged culturing yeast saccharomyces cerevisiae liquid 4%-8% step by step and ferment altogether, the 30h-32h fermentation culture of fermenting altogether finishes, go out bent oven dry then, bake out temperature is controlled at 35 ℃-45 ℃, in earlier stage (24h) inner control is at 35-40 ℃, then bake out temperature is transferred to 40-45 ℃ and just can goes out song to song moisture 10%-15%, pulverize after going out song, the product temperature is no more than 55 ℃ during pulverizing, after the song oven dry is pulverized, promptly obtain the diastatic fermentation distiller's yeast (or tropina diet) of pre-treatment plant materials material.And then the plant materials of above-mentioned adding wheat bran is added 2.2-4 times of water of its quality is 4-5 with above-mentioned alkali aqueous solution its pH value that neutralizes, insert behind the diastatic fermentation distiller's yeast of its solids content 0.6%-1.2% behind 28 ℃ of-35 ℃ of bottom fermentation 4d-6d, promptly make the maturing fermentation mash, mash obtains azeotropic alcohol through sterile filtration rectifying, and this alcohol can obtain 99.9% spirituous solution through dehydration.The solid substances that leaches obtains pre-treatment plant materials diastatic fermentation distiller's yeast or tropina diet after 35-45 ℃ of drying.Above-mentioned technological process also can be carried out semi continuous operation or operate continuously.The use of its technological process organic nitrogen source material, other organic nitrogen source material admixtion of also available and the equivalence of wheat bran amount nutrition balance.

2, cultivate at 28 ℃ of-35 ℃ of bottom fermentations according to claim 1 is said, it is characterized in that: (in the 24h) cultivates the product temperature at 28 ℃-30 ℃ in earlier stage, mid-term, (24-60h) product temperature were at 30 ℃-35 ℃, later stage, (60-72h) control product temperature were at 30 ℃-33 ℃, and beginning ventilation for the first time after 24h hour simultaneously, each ventilation time is no more than 3min.Turn over the bent time and carry out after cultivating 40-42h, turn over bent number of times at 3-4 times, the water content of fermentation materials is controlled at 50%-60%.

3, said behind 28 ℃ of-35 ℃ of bottom fermentation 4d-6d according to claim 1, promptly make ripe mash, it is characterized in that: carry out diastatic fermentation product temperature control in earlier stage at 28 ℃-30 ℃, this yeast phase is 24h, the temperature control of lord ferment period product is at 30-35 ℃, and should stir mash 1-2 time every day during this, and keep 2-3d, the product temperature control of secondary fermentation phase is at 30 ℃-33 ℃, and the time need be kept 1-2d.

4, it is characterized in that according to said semi continuous operation of claim 1 or operate continuously: its single-stage periodical operation treating processes is finished the resulting ripe mash in back not exclusively send to distillation, use as the new bacterial classification song that adds the diastatic fermentation of pre-treatment plant materials material and carry out the repetition periodical operation but stay sub-fraction.When its operate continuously is handled, be that the prior fermentation mash that is obtained in the treating processes its intermittence is arrived the next stage reactor through overflow (or conveying) on the basis of adding virgin material continuously, carry out the fermentation in mid-term again, Deng finish the fermentation in mid-term to the mash of second stage reactor after, overflow again (or conveying) is carried out later stage fermentation in the next stage reactor, after finishing third stage fermentation, get final product overflow (or conveying) and filter rectification process and processed to distillation workshop section.The continuous proportioning adding mode of the diastatic fermentation distiller's yeast of this continuous conversion fermenting process is that its fermentation liquid refluxes.