CN105637083A

CN105637083A - System for management of yeast to facilitate the production of ethanol

Info

Publication number: CN105637083A
Application number: CN201380078548.1A
Authority: CN
Inventors: 尼拉康塔姆·V·纳雷德拉纳斯; 大卫·查尔斯·卡尔森
Original assignee: Baud Research Co
Current assignee: Baud Research Co; Poet Research Inc
Priority date: 2013-05-28
Filing date: 2013-05-28
Publication date: 2016-06-01
Also published as: BR112015030015A2; EP3004325A4; WO2014193344A1; BR112015030015B1; BR112015030015B8; EP3004325A1

Abstract

A system and method for managing an ethanologen for use in biorefinery is disclosed. The method for propagating ethanologen for use in the production of a fermentation product from biomass comprises the steps of providing a medium for propagation of ethanologen; supplying a first cell mass of ethanologen to the medium; supplying xylose to the medium as a carbon source for the ethanologen; and maintaining the medium comprising the first cell mass of ethanologen at a pH of between about 5.0 and 6.0 and at a temperature of between about 26 and About 37 degrees Celsius so that the first cell mass of ethanologen is propagated into a second cell mass of ethanologen. The second cell mass of ethanologen is larger than the first cell mass of ethanologen. The system for propagating ethanologen for use in the production of a fermentation product from biomass comprises a first stage comprising a first vessel configured to maintain a medium comprising ethanologen; a second stage comprising a second vessel configured to maintain a medium supplied from the first stage; a source of xylose to be provided to the medium as a carbon source for ethanologen in the first stage; and a source of xylose to be provided to the medium as a carbon source for the ethanologen in the second stage. The ethanologen has a first cell mass when supplied to the first stage and the ethanologen has a second cell mass when supplied from the first stage to the second stage and the ethanologen has a third cell mass when supplied from the second stage. The second cell mass is at least 200 times larger than the first cell mass The third cell mass is at least 20 times larger than the second cell mass.

Description

For managing yeast to promote the system of alcohol production

The cross reference of related application

This application claims the priority of the U.S. Provisional Application that the title submitted on March 3rd, 2009 is " PropagationofPentoseMetabolizingYeastCells " and series number is 61/157,151, and be incorporated by reference herein.

The application relates to following application, and is incorporated by reference herein: the title that (a) on March 3rd, 2010 submits to is " SystemforPre-TreatmentofBiomaesfortheProductionofEthanol " and U. S. application that application number is (Atty.DocketNo (attorney docket) .P1841250.1); (b) title that on March 3rd, 2010 submits to is " SystemforFermentationofBioraassfortheProductionofEthanol " and U. S. application that application number is (Atty.DocketNo (attorney docket) .P1841280.1).

Technical field

The application relates to produce cellulosic ethanol and reclaim the system of other biological product. The application further relates to for managing yeast to promote the system of alcohol production. The application is further to being used for breeding producing and ethanol material for the method producing tunning from biomass.

Background technology

Ethanol can produce from cereals raw material (such as Semen Maydis, Sorghum vulgare Pers./chinese sorghum, Fructus Hordei Vulgaris, Semen Tritici aestivi, Semen sojae atricolor etc.), saccharide is (such as from Caulis Sacchari sinensis, Radix Betae etc.) and biomass (such as lignocellulosic material, such as switchgrass, corncob and straw, timber or other plant material).

Biomass include going for being directly used as fuel/energy source or being used as to be processed in bio-refineries (such as ethanol plant) plant material of other biological goods (such as, bio-fuel, such as cellulosic ethanol). Biomass can include, such as, during harvesting corn seed or the corncob obtained afterwards and straw (such as, stem, leaf), fiber from corn kernel, switchgrass, farm or agricultural residue, wood flour or other wood waste fiber and other plant materials (plantation be used for being processed into biological product or for other purposes). For carrying out using or processing, from field results biomass collection, and it is transported to the place using or processing.

From the conventional ethanol factory of Maize Production ethanol, ethanol prepares from starch. Corn kernel is cleaned and grinds, to be ready for the starch-containing material processed, (also corn kernel can be carried out fractional distillation, so that starch-containing material (such as endosperm) separates with other material (such as fiber and plumule)). With water starch-containing material carried out pulp and liquefy, converting the sugars into the fermentation reaction of ethanol with the saccharification react and producing and ethanol material (ethanologen, such as yeast) promoting Starch Conversion saccharogenesis (such as glucose). The product (i.e. tunning) of fermentation reaction is medicated beer, and it includes the liquid component containing second alcohol and water and water soluble ingredient and the solid constituent containing the particle matter that do not ferment (and other materials). Tunning is delivered to Distallation systm. In Distallation systm, described tunning carries out distilling and being dehydrated into ethanol. By residue (such as, full distiller grains) include water, water soluble ingredient, oil and non-fermentation solid (i.e. beer solids component after all ethanol of basic removing, it can be dried to distiller's dried grain (DDQ) and sell as animal feed product). Other joint product can also be reclaimed, for instance syrup (with oil contained in syrup) from distiller grains. Can carry out in the factory processing to reuse by distilling the water removed from tunning,

Being used for from the bio-refineries of biomass production ethanol, ethanol produces from ligno-cellulosic materials. Lignocellulose biomass generally includes cellulose, hemicellulose and lignin. Cellulose (a kind of glucosan) is the polysaccharide comprising hexose (C6) sugar monomer (such as glucose) that straight chain is connected. Hemicellulose is branched polysaccharides, and it can include some different pentose (C5) sugar monomers (such as xylose and arabinose) and a small amount of hexose (C6) sugar monomer side chain.

Preparing biomass, thus making saccharide in described ligno-cellulosic materials (as from the glucose of hemicellulose with from cellulosic xylose) become upstate, can ferment for tunning, it is possible to therefrom reclaim ethanol. Tunning after fermentation is sent to Distallation systm, at this by distilling and dehydration recovery ethanol. In the process that biomass processing is ethanol, it is also possible to the other biological product as by-product or joint product is reclaimed, for instance lignin and organic acid. Determine how more effectively to prepare and process biomass for alcohol production, depend on the composition of the source of biomass, type or biomass. Dissimilar or separate sources biomass are likely to be of different character and composition (relative quantity of such as cellulose, hemicellulose, lignin and other composition). Such as, the composition of wood flour will differ from the composition of corncob or switchgrass.

There is provided and be advantageous for for the system producing cellulosic ethanol, provide a kind of for managing yeast to promote the system of alcohol production also advantageously. Further, it is provided that the system of a kind of feature that one or more promotion efficiency from biomass production cellulosic ethanol and output increased can be provided, it is also advantageous that.

Summary of the invention

The present invention relates to for breeding producing and ethanol material for the method producing tunning from biomass. The method comprises the following steps: provide the culture medium for breeding producing and ethanol material; The first cell mass of producing and ethanol material is supplied to described culture medium; The xylose carbon source as producing and ethanol material is supplied to described culture medium; And comprise described in keeping the pH of culture medium of described producing and ethanol material the first cell mass between about 5.0 to 6.0 and temperature between about 26 to about 37 degrees Celsius so that the second cell mass that the breeding of described producing and ethanol material the first cell mass is producing and ethanol material. Described producing and ethanol material the second cell mass is more than described producing and ethanol material the first cell mass.

The invention still further relates to for breeding producing and ethanol material for the method producing tunning from biomass. The method comprises the following steps: provide the culture medium for the breeding of producing and ethanol material; Producing and ethanol material the first cell mass is supplied to described culture medium; Additive is provided to described culture medium; The composition taking from described biomass is provided, as the carbon source of described producing and ethanol material to described culture medium; And comprise described in keeping the pH of culture medium of described producing and ethanol material the first cell mass between about 5.0 to 6.0 and temperature between about 26 to about 37 degrees Celsius so that the second cell mass that the breeding of described producing and ethanol material the first cell mass is described producing and ethanol material. Described producing and ethanol material the second cell mass is more than described producing and ethanol material the first cell mass. Described biomass include cellulosic material. Described cellulosic material includes at least one in corncob, corn plant skin, corn plant leaf and corn plant bar. Described composition obtains from described cellulosic material; Described composition includes pentose. Described pentose includes the carbon source for described producing and ethanol material. Described producing and ethanol material includes to be the yeast cells of ethanol by pentose fermentation.

The invention further relates to breeding producing and ethanol material for the system producing tunning in fermentation system from biomass. This system includes: the first stage, including for keeping the first container comprising the culture medium of described producing and ethanol material; Second stage, including for keeping the second container of culture medium provided by the described first stage; Supply the described culture medium xylose source as producing and ethanol material carbon source in the described first stage; With supply the described culture medium xylose source as producing and ethanol material carbon source in described second stage. When being supplied to the described first stage, described producing and ethanol material has the first cell mass; When being supplied to described second stage from the described first stage, described producing and ethanol material has the second cell mass; And when providing from described second stage, described producing and ethanol material has the 3rd cell mass. Described producing and ethanol material keeps the pH of described culture medium in described first container between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that can be bred as described second cell mass; Described producing and ethanol material keeps the pH of described culture medium in described second container between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that can be bred as described 3rd cell mass. Described second cell mass is than big at least 200 times of described first cell mass. Described 3rd cell mass is than big at least 20 times of described second cell mass.

Accompanying drawing explanation

Figure 1A is the perspective view of the bio-refineries including cellulosic ethanol production equipment.

Figure 1B is the perspective view of bio-refineries including cellulosic ethanol production equipment and the alcohol production equipment based on Semen Maydis.

Fig. 2 receives and prepares the biomass schematic diagram for the system of cellulosic ethanol production equipment.

Fig. 3 is the theory diagram of the system for producing ethanol from biomass.

Fig. 4 A, 4B and 4C are the theory diagrams of the processing system of the composition for producing ethanol gained from biomass.

Fig. 5 A and 5B is the schematic flow sheet of the system for producing ethanol from biomass.

Fig. 6 A be for preparing, the theory diagram of the device of pretreatment and separating biomass.

Fig. 6 B is the perspective view for pretreatment and the device of separating biomass.

Fig. 7 A is the flow process theory diagram of the system for producing tunning from liquid component (C5).

Fig. 7 B is the flow process theory diagram of the system in an example embodiment.

Fig. 7 C is the flow process theory diagram of the system in an example embodiment.

Fig. 8 A is the flow process theory diagram of the system in an example embodiment.

Fig. 8 B is the flow process theory diagram of the system in an example embodiment.

Fig. 9 is the theory diagram of the system in an example embodiment.

Figure 10 A show the line graph of producing and ethanol substance migration xylose growth.

Figure 10 B show the line graph of producing and ethanol substance migration xylose growth.

Figure 11 A show the line graph of producing and ethanol agent growth.

Figure 11 B show the line graph of producing and ethanol agent growth.

Figure 12 A show the line graph of producing and ethanol substance migration xylose growth.

Figure 12 B show producing and ethanol substance migration xylose and the line graph of bio liquid component growth.

Figure 12 C show the line graph of the bio liquid component growth after producing and ethanol substance migration xylose and process.

Figure 13 A show in the second stage of two benches breeding, the line graph of producing and ethanol substance migration xylose growth.

Figure 13 B show in the second stage of two benches breeding, the line graph of producing and ethanol substance migration xylose growth.

Figure 14 A show in the second stage of two benches breeding, the line graph of the initial xylose supply growth of producing and ethanol substance migration.

Figure 14 B show in the second stage of two benches breeding, the line graph of the continuous xylose supply growth of producing and ethanol substance migration.

Figure 15 A show the line graph of the xylose consumption when ventilation and stuffiness of the producing and ethanol material and alcohol production.

Figure 15 B show the line graph of the producing and ethanol material growth when ventilation and stuffiness.

Figure 16 A is the chart of producing and ethanol substance migration xylose growth.

Figure 16 B is the chart of producing and ethanol substance migration xylose growth.

Figure 16 C is the chart of producing and ethanol substance migration xylose growth.

Table 1A and 1B lists in example embodiment and exemplary embodiments, comprise the composition of the biomass of the cellulosic plant material from corn plant.

Table 1A and 1B list in example embodiment and exemplary embodiments, the liquid component composition of preprocessing biomass.

Table 1A and 1B list in example embodiment and exemplary embodiments, the solid constituent composition of preprocessing biomass.

Table 4 lists the growth medium composition in embodiment 1-4.

Table 5A and 5B lists the use result of system in embodiment 1A.

Table 6A and 6B lists the use result of system in embodiment 1B.

Table 7A, 7B and 7C list the use result of system in embodiment 2.

Table 8A and 8B lists the use result of system in embodiment 3.

Table 9A and 9B lists the use result of system in embodiment 4.

Table 10A and 10B lists the use result of system in embodiment 5.

Table 11A to 11C lists the use result of system in embodiment 6.

Detailed description of the invention

Referring to Figure 1A, it as shown is a kind of bio-refineries for producing ethanol from biomass.

In an exemplary embodiment, bio-refineries is for producing ethanol from the biomass of lignocellulosic material form (vegetable material (such as corncob and corn straw straw) as from corn plant). Lignocellulosic material, as included cellulose (can therefrom obtain C6 sugar such as glucose) and/or hemicellulose (can therefrom obtain C5 sugar such as xylose and arabinose) from the ligno-cellulosic materials of corn plant.

As shown in Figure 1A, bio-refineries includes for carrying biomass and preparing biomass for the region being supplied to cellulosic ethanol production facility. Described cellulosic ethanol production facility includes for biomass being prepared, pretreatment and process are the device suitable in the processed biomass being fermented into tunning fermentation system. This facility includes the Distallation systm carrying out distilling and being dehydrated into ethanol for described tunning. As shown in Figure 1A, bio-refineries may also include waste treatment system (for including anaeroic digestor and electromotor shown in figure). In other washability embodiment, described waste treatment system can include for carrying out other equipment processing, process and reclaiming in cellulosic ethanol production process, such as solid/refuse fuel boiler, anaeroic digestor, aerobic digestive appartus or other biochemistry or chemical reactor.

As shown in Figure 1B, in an exemplary embodiment, bio-refineries can include cooperating together cellulosic ethanol production facility (it produces ethanol from ligno-cellulosic materials and corn plant composition) and the ethanol production facilities (its starch contained therein from the endosperm composition of corn kernel produces ethanol) based on Semen Maydis. As shown in Figure 1B, cooperated together by two ethanol production facilities, it is possible to share some factory system, such as, for the system of dehydration, storage, degeneration and ethanol transport, by the Fa electricity Xi Tong factory management of energy/fuel conversion energy and control system, and other system. Corn fiber (constituent of corn kernel) can prepare corn kernel (such as grinding needs in based on the ethanol production facilities of Semen Maydis, by sorting) time obtains, and it can be supplied to cellulosic ethanol production facility as raw material. The fuel obtained from cellulosic ethanol production facility or the energy, such as methane or lignin, can be used for cooperating together to two facility or one of them the offer energy. In other alternative embodiment, bio-refineries (such as cellulosic ethanol production facility) can cooperate together with other type of factory and facility, for instance power plant, waste disposal facilities, timber mill, the facility of paper mill or process agricultural product.

Referring to Fig. 2, it as shown is the system of biomass for preparing to be transported to bio-refineries. Biomass preparation system can include receiving for biomass/unloading, clean (namely removing impurity), grinding (namely milling, reduce or densification) and transport and be transported to the device that factory carries out processing. In an exemplary embodiment, the biomass of corncob and straw form can be transported to bio-refineries and store (such as with forms such as bag, heap or casees), and managed for this facility. In a preferred embodiment, biomass can include corncob and corn straw and other material of at least 20% to 30% (by weight). In other one exemplary embodiment, the preparation system of bio-refineries may be used for preparing all various biomass (i.e. vegetable material), is processed to and is processed into ethanol and other biological product for factory.

Referring to Fig. 3, it as shown is the schematic diagram of cellulosic ethanol production facility. In a preferred embodiment, to including being prepared from the biomass of the vegetable material of described corn plant and cleaning in preparation system. After preparation, in pretreatment system, biomass and water are mixed into slurry, and carry out pretreatment. In pretreatment system, biomass are decomposed (such as by being hydrolyzed), are beneficial to be separated into liquid component (such as containing the stream of C5 sugar) and solid constituent (such as comprising the cellulosic stream that can therefrom obtain C6 sugar). Liquid component (C5 stream) containing C5 sugar and the solid constituent (C6 stream) containing C6 sugar can process (if suitable) and fermentation in fermentation system. Tunning from fermentation system is supplied to the Distallation systm reclaiming ethanol.

As shown in Fig. 3 and Fig. 4 A, the composition removed when processing C5 stream can carry out processing or process to reclaim by-product, for instance organic acid and furfural. As shown in Fig. 3 and Fig. 4 B, the composition removed when processing C6 stream, such as lignin or other composition, can process or be processed into biological product or fuel (if lignin is for solid fuel fired boiler, or by processing methane that residue/removing substances (such as acid and lignin) produces for anaeroic digestor). Such as Fig. 4 A, shown in 4B and 4C, the composition that during with C5 stream and C6 stream or the biomass processes in one of them and production ethanol, (or during distillation) removes, it is possible to be processed to biological product (such as, by-product or joint product) or recycle or recycling. As shown in Figure 4 C, remove composition (such as distiller grains or remove solid) from what Distallation systm obtained or remove composition (as the solid removed and particulate matter during from distillation pre-treatment tunning, it can include residual lignin etc.), can process or be processed into biological product or fuel (methane such as, produced in anaeroic digestor).

In a preferred embodiment, biomass comprise the vegetable material from corn plant, for instance corncob, crust and leaf and stem; The constituent of vegetable material (i.e. cellulose, hemicellulose and lignin) is substantially as shown in table 1A and IB. In a preferred embodiment, vegetable material includes corncob, skin/leaf and stem (after namely cleaning/remove impurity), such as, vegetable material can include (by weight) and be up to 100% cob, skin/the leaf of maximum 100%, the cob of about 50% and the skin/leaf of about 50%, the cob of about 30% and the skin/leaf of about 50% and about 20% stem, or other any combination from the cob of corn plant, skin/leaf and stem. In Table 1A. In an exemplary embodiment, corn stalk includes the first half or 3/4ths parts of bar. In one alternate embodiment, described lignocellulosic plants material can include the fiber (form being such as combined) from corn kernel with other vegetable material. Table 1B provides typical case and the desired extent of the representative composition of the biomass being considered to include the ligno-cellulosic materials from described corn plant. In an exemplary embodiment, lignocellulosic plants material (from corn plant) in biomass comprises the cellulose of (by weight) about 30% to 55%, hemicellulose is about 20% to 50%, and lignin is about 10% to 25%. In a particularly preferred embodiment, lignocellulosic plants material (the cob of corn plant in biomass, skin/leaf and stem portion) comprise the cellulose of (by weight) about 35% to 45%, hemicellulose is about 24 to 42%, and lignin is about 12% to 20%. In a particularly preferred embodiment, preprocessing biomass will produce the liquid component comprising the xylose being not less than 1.0% (by weight) and the solid constituent comprising the cellulose (therefrom can obtain glucose) being not less than 45% (by weight).

Referring to Fig. 5 A and 5B, it as shown is the one exemplary embodiment of system for producing wood-based composites. As shown in Figure 5 A and 5B, biomass carry out pretreatment in pretreatment system, are then peeled off into liquid component and solid constituent.

In a preferred embodiment, in pretreatment system, add acid to ready biomass, to promote biomass decomposition, to be separated into liquid component (C5 stream therefrom recyclable fermentable C5 sugar) and solid constituent (C6 flows, therefrom recyclable fermentable C6 sugar). In a preferred embodiment, acid can in operating condition (the i.e. acid concentration determined, pH, temperature, the time, pressure, solid concentration, flow velocity, supply process water or steam etc.) under add in the biomass in reaction vessel, and biomass can be stirred in reaction vessel/mix, to promote the decomposition of biomass. In the exemplary embodiments, it is possible to add acid to biomass, for instance sulphuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid etc. (or preparation/mixture of acid). In a particularly preferred embodiment, with process in, to biomass add sulphuric acid.

Liquid component (C5 stream) includes water, will can be used for being fermented into the sugar (such as xylose, arabinose and glucose) of the dissolving of ethanol, acid and be recovered from other soluble component of hemicellulose. (table 2B provides typical case and the desired extent of thinking that the representativeness of the biomass including the ligno-cellulosic materials from described corn plant forms. ) in an exemplary embodiment, liquid component can comprise about the solid (i.e. suspension/residual solid, for instance the hemicellulose of partial hydrolysis, cellulose and lignin) of 5% to 7%. In a particularly preferred embodiment, liquid component comprises the xylose (by weight) of at least 2% to 4%. In other one exemplary embodiment, liquid component is not less than the xylose (by weight) of 1% to 2% by comprising. Table 2A and 2B lists in exemplary and representational embodiment, the composition of the liquid component of pretreated biomass (as from the ready biomass shown in table 1A and 1B).

Solid constituent (C6 stream) comprises water, acid and solid (as can be therefrom obtained saccharide (such as glucose) for the cellulose being fermented into ethanol) and lignin. (table 3B provides typical case and the desired extent of the representative composition of the biomass including the ligno-cellulosic materials from described corn plant. ) in an exemplary embodiment, solid constituent can include about the solid (by weight) (after separation) of 10% to 40%; In a particularly preferred embodiment, solid constituent includes the solid (by weight) of about 20% to 30%. In a preferred embodiment, the solid in solid constituent includes the cellulose being not less than 30%, and solid constituent can also include other solvable saccharide (such as glucose and xylose). Table 3A and 3B lists the composition of the solid constituent of preprocessing biomass in exemplary and representational embodiment (as from the ready biomass shown in table 1A and 1B).

In preprocessing process, the severe degree of operating condition (such as pH, temperature and time) may result in the formation of the composition suppressing fermentation. Such as, under certain conditions, the dehydration of C5 sugar (such as xylose or arabinose) can cause furfural and/or the generation of Hydroxymethylfurfural (HMF). Acetic acid is likely to formation, for instance, preprocessing process discharges during cellulose decomposition acetic acid. Sulphuric acid be introduced into ready biomass to promote pretreatment, if but be not removed or neutralize, it is also possible to suppress fermentation. In an exemplary embodiment, by regulating the condition (such as pH, temperature and time) of pretreatment, it is possible to reduce or control the formation of suppressant additive; In other one exemplary embodiment, the component of pretreated biomass can process remove suppressant additive (or other undesirable material) or reduce its concentration further.

After Fig. 5 ' A and 5B, pretreatment and separation, C5 stream and C6 stream are respectively processed; As it can be seen, C5 stream and C6 stream separately can process before co-fermentation (C5/C6 fermentation as shown in Figure 5A), or separately process, including separately fermentation (independent C5 fermentation and C6 fermentation as shown in Fig. 5 B).

The C5 of biomass can be flowed (liquid component) to process, to remove composition (the such as furfural hindering high-efficiency fermenting from C5 saccharic composition, Hydroxymethylfurfural, sulphuric acid and acetic acid) and it is likely to azymous residual lignin (or other material), thus can by saccharide (such as xylose, arabinose and other saccharides, for instance glucose) it is used for fermenting. C5 sugar in C5 stream can also carry out concentrating improving fermentation efficiency (such as, to improve the titre of ethanol for distillation).

C6 stream (solid constituent) of described biomass can carry out the C6 sugar processing to obtain can be used for fermentation. In a preferred embodiment, it is possible to be hydrolyzed (such as enzyme hydrolysis) C6 sugar to obtain in cellulose; The lignin that can also carry out processing removing in C6 stream and other non-fermented composition (or to remove the component being likely to suppress high-efficiency fermenting, such as residual acid or acid etc.).

In the one exemplary embodiment shown in Fig. 5 A, C5 stream and C6 stream after pretreatment and separation can separately process, and merge (such as slurry) after treatment to carry out co-fermentation in fermentation system with generation C5/C6 tunning from available sugar (such as xylose and glucose); C5/C6 tunning (if there being process, then after treatment) can be supplied to Distallation systm to reclaim ethanol (such as, by distilling and dehydration). In one exemplary embodiment shown in figure 5b, C5 stream and C6 stream can each carry out fermenting respectively and distillation processes (if there being process, then after treatment) to produce ethanol. In arbitrary preferred embodiment, it is possible to the selection using suitable fermenting organism (producing and ethanol material) producing and ethanol material in fermentation system can based on various considerations, for instance the main species of sugar contained in sauce. Ethanol from C5 stream and C6 stream dehydration and/or degeneration can individually or merging carry out.

Fig. 6 A and 6B illustrates in an exemplary embodiment, for preparing, the segregation apparatus of pretreatment and separation lignocellulose biomass. As it can be seen, biomass are prepared in grinder (such as pulverizer or other suitable device or flour mill). Ready biomass carry out in reaction vessel (or reaction vessel group), wherein adopt the ready biomass of predetermined concentration (or pH) and acid/water and other operating condition. As shown in Fig. 6 B, pretreated biomass can be separated into liquid component in centrifuges, and (C5 flows, mainly comprise liquid and some solids) and solid constituent (C6 stream, including liquid and solid, such as lignin and cellulose, therefrom can pass through to process further obtain glucose).

As shown in Fig. 7 A and 7B, liquid component (C5 stream) is introduced in process system. In this process system, C5 stream can pass through filtration treatment, and/or by can be used for the liquid component of the sugar of fermentation including after being condensed into process. The liquid component processed is supplied in fermentation system, to produce tunning (it comprises ethanol) by producing and ethanol material (namely sugar fermentation is produced tunning by organism such as yeast).

In a preferred embodiment, producing and ethanol material includes the yeast that atom transgenic recombinant Saccharomyces cerevisiae is derivative. In a particularly preferred embodiment, producing and ethanol material is the Wine brewing yeast strain (one is derived from the Recombinant yeast of organism described in the United States Patent (USP) 7622284 as transferred Nei Daerke company of imperial family (RoyalNedalcoB.V.)) transforming as and xylose and glucose can being converted into ethanol.

With reference to Fig. 7 B, provide a kind of propagating system for producing and ethanol material (being yeast shown in figure). In this propagating system, growth medium is provided (such as to yeast, water and carbon source, such as sugar) and additive is (such as, nutrient etc.) promote that the abundant of yeast (i.e. yeast cells group) grows, for inoculation (namely providing yeast-inoculated body) to fermentation system.

In an exemplary embodiment, the growth medium for propagating system includes, for instance, aseptic yeast extract-peptone culture medium, as the xylose of carbon source and other additives (such as nutrient). The additive being supplied to producing and ethanol material can include antibiotic, supplement or auxiliary enzymes, nutrient or the composition to organism offer nutrition or other benefit. Nutrient can comprise yeast extract, carbamide, Diammonium phosphate (DAP), magnesium sulfate, zinc sulfate or other salt etc. In an exemplary embodiment, yeast-inoculated body cultivates about 17 hours when the temperature of about 30 degrees Celsius and pH about 5.5. In an alternative embodiment, in order to cultivate (inoculation) yeast in propagating system, its temperature may remain in the scope of about 28 to 32 degrees Celsius, and its pH value may remain in the scope of about 5.2 to 5.8, or the time of at least 12 hours.

Referring to Fig. 7 C and Fig. 9, it as shown is the one exemplary embodiment of the two benches propagating system of a producing and ethanol material (for yeast shown in figure). As it is shown in figure 9, yeast culture can grow up to the initial yeast inoculum that can be used for being supplied to the propagating system first stage.

In the first reproductive stage, initial yeast inoculum is transferred in bigger container and dilutes (such as, 250 times). In this embodiment, initial yeast inoculum can provide with part C5 stream (namely including the liquid component of other saccharides of C5/) and water together with additive (such as nutrient) and fresh yeast. In a preferred embodiment, in the first reproductive stage, yeast cultivates about 24 hours when including temperature about 30 degrees Celsius and pH about 5.5. In other one exemplary embodiment, it is possible to be maintained at about in the temperature range of 26 to 37 degree and the time of at least 24 hours within the scope of the pH of about 3.5 to 6.5. In the second reproductive stage, the yeast-inoculated body from the first reproductive stage is transferred in bigger container and dilutes (such as, 10 times). In this embodiment, can provide together with additive (such as nutrient) and fresh yeast from the yeast-inoculated body of described first reproductive stage with part C5 stream (namely including the liquid component of other saccharides of C5/) and water. . In a preferred embodiment, in the second reproductive stage, yeast cultivates about 24 hours when including temperature about 30 degrees Celsius and pH about 5.5. In other one exemplary embodiment, it is possible to be maintained at about in the temperature range of 26 to 37 degree and the time of at least 24 hours within the scope of the pH of about 3.5 to 6.5.

Tunning (is alternatively referred to as medicated beer or fermentation liquid, or be called comprise medicated beer or fermentation liquid) comprise second alcohol and water and unfermentable material (such as, any unfermentable sugar) and not fermentable material (lignin such as remained and other solids). Tunning also includes once for the granular producing and ethanol material (i.e. yeast cells) producing ethanol and other component produced by fermentation system, for instance, glycerol (tunning) and acetic acid.

Such as Fig. 8 A and 8B if so, in an exemplary embodiment, it is also possible to a kind of process system for tunning is provided. This process system can include tunning is separated into liquid component (tunning namely processed, it mainly includes second alcohol and water) and solid constituent (it mainly includes solid matter, for instance producing and ethanol material/yeast cells). In a preferred embodiment, as shown in, in Fig. 8 B, described tunning being separated into liquid component and solid constituent, can carry out in centrifuges. As shown in Figure 8 B, fermentation system recycling (i.e. recycling in fermentation tank) can be supplied to together with extra or fresh yeast cell (as required) including the processed solid constituent of yeast cells. Yeast cells can process in yeast cells process system. This yeast cells processes system can include washing separated yeast cell, uses it for the recirculation in fermentation tank or tunning stream subsequently.

In arbitrary preferred embodiment, this yeast propagation system supports that yeast is with suitable growth rate for the suitable yeast cells group being provided to fermentation system. In a preferred embodiment, this system allows yeast to utilize xylose to grow as growth carbon source. In a particularly preferred embodiment, this system allows to the selective growth using xylose as the yeast (yeast can bred in the culture medium containing xylose) of carbon source, even if there is other yeast (namely as pollutant); In the xylose culture medium as sole carbon source (namely not comprising a large amount of glucose) is provided, the yeast that xylose is bred as carbon source can be utilized to breed, and cannot utilize that xylose breeds as carbon source other/pollutant yeast (such as the usual more conventional yeast of breeding in the culture medium containing glucose) will be unable to identical speed breeding (or not breeding). In a particularly preferred embodiment, xylose and glucose fermentation can be all ethanol by yeast. In a preferred embodiment, growth medium and environment that this yeast propagation system provides make yeast that sugar converts (such as xylose) yeast cells group rather than ethanol, such as, when such as ventilation (referring to Figure 15 A and 15B), or during permission yeast propagation, tolerate the sugar (such as xylose) (referring to Figure 16 A and 16B) of higher concentration. In arbitrary preferred embodiment, this system will promote that yeast cells group is effectively grown to the inoculum of the fermentation system that can be supplied to bio-refineries. In a particularly preferred embodiment, yeast cells group will increase about 200 to 500 times in the first phase, and increase about 20 to 40 times in second stage.

***

One exemplary embodiment according to described yeast propagation system, carried out a series of embodiment (as shown in the figure, for instance, Fig. 7 B, in 7C and 9), to assess the growth under various conditions of producing and ethanol material. The producing and ethanol material used in an embodiment is that a kind of saccharomyces cerevisiae yeast strain that xylose and glucose can be converted to ethanol (is derived from such as imperial family Nei Daerke company (RoyalNedalcoB.V.,) the transgenic yeast of the organism described in U.S. Patent number 7622284, for instance bacterial strain RWB218; Bacterial strain RN1001; With bacterial strain RN1014). Shown in the data of embodiment such as Figure 10 A to 16C and table 5A to 11C. Table 4 lists the embodiment 1A composition (including the additive added, such as nutrient) to the growth medium in embodiment 4.

Using sterilised yeast extract-peptone (YP) culture medium (yeast extract is every liter of culture medium 12.5 grams, and peptone is 10 grams every liter) is Initial Inoculum (yeast-inoculated body) by producing and ethanol material producing and ethanol material. Inoculum is cultivated under 30 degree Celsius for about 17 to 18 hours. Inoculum used: culture medium ratio is 1:250. Use a pump to control xylose delivery rate. The pH of this culture medium is maintained within 5.5 (by adding the potassium hydroxide solution of 45 weight %). It is periodically subject to yeast growth (cell mass), sugar, the sample analysis of organic acid and ethanol. Measure the optical density (spectrophotometer, the 600nm) index as the yeast amount (i.e. cell mass) in sample. (adopt other composition of high-efficient liquid phase chromatogram technique analysis. )

Embodiment 1A

In embodiment 1A, described propagating system is used to regulate the impact on producing and ethanol agent growth to evaluate pH value. This producing and ethanol material is yeast (bacterial strain RN1001). Sample is prepared at two independent reaction vessels. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body. Sample also includes the xylose that concentration is 30 gram per liter (culture medium). The pH regulator of the sample in each reaction vessel is to 5.5. Temperature in each reaction vessel is maintained at about 30 degree Celsius. Yeast is bred 54 hours in each reaction vessel. In one of reaction vessel, pH keeps (adjustment) to be 5.5; In other reaction vessels, pH is not for be adjusted. After 54 hours, adding xylose solution (by weight 50%) and other nutrient, total amount is about 60 g/l of xyloses. The breeding of yeast continues 66 hours again. In one of reaction vessel, pH is kept (adjustment) is 5.5; In other reaction vessels, pH is not for be adjusted. Yeast growth (dry weight) in test sample, concentration of alcohol, xylose concentration and optical density (OD₆₀₀), to assess xylose. It has been observed that pH value is maintained at the sample of 5.5 creates more yeast than in the sample (in described experiment condition) not regulating pH. Described result is shown in Figure 10 A and 10B and table 5A and 5B.

Embodiment 1B

In embodiment 1A, use described propagating system to evaluate the different xylose feed speed impact on the growth of producing and ethanol material. This producing and ethanol material is yeast (bacterial strain RN1001). Sample is prepared at two independent reaction vessels. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body. Sample also includes the xylose that concentration is 20 gram per liter (culture medium). Temperature in each reaction vessel is maintained at about 30 degree Celsius. Yeast is bred 24 hours in each reaction vessel. After 24 hours, in 46 hours, by xylose solution (by weight 50%, and other nutrients) send into one of reaction vessel with the speed of 0.14 milliliter per minute, the xylose total amount added is 90 grams every liter, and send into other reaction vessel with the speed of 0.28 milliliter per minute, the xylose total amount added is every liter of about 100 grams of xyloses. The breeding of yeast continues 50 hours again. Yeast growth (dry weight) in test sample, concentration of alcohol, xylose concentration and optical density (OD₆₀₀), to assess xylose. It has been observed that yeast growth speed is similar in sample, enter the speed (in described experiment condition) of sample independent of xylose. Described result is shown in Figure 11 A and 11B and table 6A and 6B.

Embodiment 2

In example 2, use described propagating system to evaluate not by the impact on producing and ethanol agent growth of the xylose source. By preparing the first bio liquid component (referring to Fig. 6 A and table 2A and 2B) with dilute acid pretreatment corncob. By processing described first liquid component with ion exchange chromatography, prepare processed bio liquid component. This producing and ethanol material is yeast (bacterial strain RN1001). Sample is prepared at three independent reaction vessels. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body. Sample also includes the xylose that concentration is 20 gram per liter (culture medium). The pH regulator of each sample to and to keep (adjustment) be 5.5. Temperature in each reaction vessel is maintained at about 30 degree Celsius. Yeast is bred 24 hours in each reaction vessel. After 24 hours, in about 22 hours, xylose solution (by weight 50%, and other nutrients) is sent in the first reaction vessel with the speed of 0.19 milliliter per minute, the xylose total amount added is about 106 grams every liter, and the breeding of yeast continues 66 hours again. After 24 hours, in about 22 hours, by xylose solution (by weight 50%, and other nutrients) send in the second reaction vessel with the speed of 0.26 milliliter per minute, hereafter biofluid composition is sent into about 2.6 hours with the speed of 2.8 milliliters per minute again, the xylose total amount added is 95 grams every liter, and the breeding of yeast continues 71 hours again. After 24 hours, in about 22 hours, by xylose solution (by weight 50%, and other nutrients) send in the 3rd reaction vessel with the speed of 0.19 milliliter per minute, hereafter biofluid composition is sent into about 2 hours with the speed of 3.0 milliliters per minute again, the xylose total amount added is 92 grams every liter, and the breeding of yeast continues 72 hours again. Yeast growth (dry weight) in test sample, concentration of alcohol, xylose concentration and optical density (600nm, OD₆₀₀), to assess xylose. It has been observed that the xylose taking from biomass (lignocellulosic plants material) can be used for breeding yeast (in described experiment condition). Described result is shown in Figure 12 A to 12C and table 7A to 7C.

Embodiment 3

In embodiment 3, use propagating system as shown in Fig. 7 C and 9 to evaluate the producing and ethanol agent growth in the two benches propagating system of different xylose concentration. This producing and ethanol material is yeast (bacterial strain RN1001). In the first phase, reaction vessel prepares sample. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body. Sample also includes the xylose that concentration is 20 gram per liter (culture medium). Yeast propagation 24 hours. After 24 hours, take the first stage sample of 300 milliliters, two samples (second stage) of each inoculation in two independent reaction vessels. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body, so that the total sample volume in each reaction vessel of second stage is 3 liters. Sample in first container of second stage also includes the xylose that concentration is 40 gram per liter (culture medium), and the sample in the second container of second stage also includes the xylose that concentration is 30 gram per liter (culture medium). Temperature in each reaction vessel is maintained at about 30 degree Celsius. Yeast is bred 12 hours in each reaction vessel. The pH regulator of each sample to and to keep (adjustment) be 5.5. After 12 hours, by xylose solution (by weight 50%, and other nutrients) with in the first reaction vessel of the speed feeding second stage of 0.26 milliliter per minute about 12 hours, the xylose total amount added was about 64 grams every liter, and the breeding of yeast continues 30 hours again. After 12 hours, by xylose solution (by weight 50%, and other nutrients) with in the second reaction vessel of the speed feeding second stage of 0.30 milliliter per minute about 12 hours, the xylose total amount added was about 64 grams every liter, and the breeding of yeast continues 30 hours again. According to observations, compared with single phase propagating system (namely using about 100 to 120 hours), two benches propagating system can reduce breeding yeast (such as, yeast cells group grows to 20 grams from about 0.04 gram in the first phase, and grows to 75 grams from about 2 grams in second stage) total time (namely using about 78 hours). See also embodiment 1A, 1B and 2. Described result is shown in Figure 13 A and 13B and table 8A and 8B.

Embodiment 4

In example 4, use the propagating system as shown in Fig. 7 C and 9 to evaluate the xylose supply impact on producing and ethanol agent growth. This producing and ethanol material is yeast (bacterial strain RN1014). In the first phase, reaction vessel prepares sample. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body. Sample also includes the xylose that concentration is 20 gram per liter (culture medium). Yeast propagation 24 hours. After 24 hours, take the first stage sample of 300 milliliters, two samples (second stage) of each inoculation in two independent reaction vessels. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body, so that the total sample volume in each reaction vessel of second stage is 3 liters. Two samples of second stage all include the xylose that concentration is 35 gram per liter (culture medium). Temperature in each reaction vessel is maintained at about 30 degree Celsius. Yeast is bred 23 hours in each reaction vessel. The pH regulator of each sample to and to keep (adjustment) be 5.5. After 23 hours, to the xylose solution (by weight 50%, and other nutrients) of the sample supply single dose in the first reaction vessel of second stage, the xylose total amount added is about 64 grams every liter, and the breeding of yeast continues 31 hours again. After 23 hours, with the speed of 0.31 milliliter per minute to the sample supply xylose solution (by weight 50% in the second reaction vessel of second stage, and other nutrients) 8.2 hours, the xylose total amount added is about 64 grams every liter, and the breeding of yeast continues 32 hours again. It has been observed that when providing extra xylose (namely after 23 hours) with the form that supplies continuously to sample, but not during single dose supply, Yeast Growth can be improved (in described experiment condition). Described result is shown in shown in Figure 14 A and 14B and table 9A and 9B.

Embodiment 5

In embodiment 5, use described propagating system to evaluate the ventilation impact on producing and ethanol agent growth. This producing and ethanol material is yeast (bacterial strain RWB218). Sample is prepared at two independent reaction vessels. Sample includes culture medium, and (such as water and additive, such as nutrient, as shown in table 4) and yeast-inoculated body. Sample also includes the xylose that concentration is 30 gram per liter (culture medium). One of reaction vessel carries out the ventilation of 5 liters per minute; Another reaction vessel stuffiness. Yeast is bred 53 hours with 32 degrees Celsius in each reaction vessel. Yeast growth (dry weight) in test sample, concentration of alcohol, xylose concentration and optical density (OD₆₀₀), to assess xylose. It has been observed that when sample is ventilated (in described experiment condition), yeast uses the xylose of supply to carry out cell mass growth rather than for producing ethanol, and saccharomycetic growth can be improved. Described result is shown in Figure 15 A and 15B and table 10A and 10B.

Embodiment 6

In embodiment 6, use described propagating system to evaluate the impact on producing and ethanol agent growth of the xylose and glucose concentration. This producing and ethanol material is yeast (bacterial strain RWB218). Sample is prepared at three independent reaction vessels. Sample includes culture medium (such as water and additive, such as nutrient, as shown in table 11A) and yeast-inoculated body. Temperature in each reaction vessel is maintained at 32 degrees Celsius. With 4.5 liters of ventilations per minute in each reaction vessel. First and second reaction vessels supply xylose with the concentration of 60 grams every liter; 3rd reactor supplies glucose with the concentration of 60 grams every liter. The pH regulator of the second and the 3rd sample in reaction vessel to and to keep (adjustment) be 5.5. Yeast in first and second reaction vessels is bred 120 hours under 32 degrees Celsius. Yeast in 3rd reaction vessel is bred 52 hours under 32 degrees Celsius. It has been observed that when sample is ventilated, yeast uses the xylose of supply to carry out cell mass growth rather than for producing ethanol, and saccharomycetic growth can be improved, regardless of xylose concentration (in described experiment condition). Shown in described result such as Figure 16 A to 16C and table 11A to 11C.

***

Embodiment (including figure and embodiment) that is disclosed herein and that describe is intended to the present invention is illustrated. The disclosed embodiments, for instance the apparatus and method of used (maybe using) and the compositions of used (maybe will use) and process, it is possible to improve and deform; All such improvement and deformation all should be regarded as and forgive within the scope of the present invention.

Table 1A

Biomass form

Table 1B

Biomass typical case and expectation composition

Table 2A

Preprocessing biomass liquid component forms

Table 2B

Preprocessing biomass liquid component composition typical range and expected range

Table 3A

Preprocessing biomass solid constituent forms

Table 3B

Preprocessing biomass solid constituent composition typical range and expected range

Table 4

Growth medium forms

^*In water

Table 5A

OD_600-600nmOptical density

NT-does not test

Table 5B

OD_600-600nmOptical density

NT-does not test

Table 6A

OD_600-600nmOptical density

NT-does not test

Table 6B

OD_600-600nmOptical density

NT-does not test

Table 7A

OD_600-600nmOptical density

NT-does not test

Table 7B

OD_600-600nmOptical density

NT-does not test

Table 7C

OD_600-600nmOptical density

NT-does not test

Table 8A

OD_600-600nmOptical density

NT-does not test

Table 8B

OD_600-600nmOptical density

NT-does not test

Table 9A

OD_600-600nmOptical density

NT-does not test

Table 9B

OD_600-600nmOptical density

NT-does not test

Table 10A

Table 10B

Table 11A

Growth medium forms

^*In water

Table 11B

OD_600-600nmOptical density

NT-does not test

Table 11C

OD_600-600nmOptical density

NT-does not test

Claims

1. the method for breeding producing and ethanol material, described producing and ethanol material, for producing tunning from biomass, said method comprising the steps of:

Culture medium for breeding producing and ethanol material is provided;

The first cell mass of producing and ethanol material is supplied to described culture medium;

Supplying xylose as carbon source to described culture medium, the cell mass for described producing and ethanol material grows;

Keep the pH of culture medium of the first cell mass comprising described producing and ethanol material between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that the second cell mass that the breeding of described producing and ethanol material the first cell mass is producing and ethanol material;

And wherein said producing and ethanol material the second cell mass is more than described producing and ethanol material the first cell mass.

2. method according to claim 1, it is characterised in that xylose is the sole carbon source provided to described culture medium.

3. method according to claim 1, it is characterised in that the step of the described culture medium of described maintenance carries out under aerobic conditions.

4. method according to claim 2, it is characterised in that the step of the described culture medium of described maintenance under anaerobic carries out.

5. method according to claim 1, it is characterised in that described biomass include ligno-cellulosic materials.

6. method according to claim 5, it is characterised in that described ligno-cellulosic materials includes at least one in corncob, corn plant skin, corn plant leaf and corn plant bar.

7. method according to claim 5, it is characterised in that the described xylose supplied to described culture medium obtains from described ligno-cellulosic materials.

8. method according to claim 1, it is characterised in that described producing and ethanol material includes to be the yeast cells of ethanol by pentose fermentation.

9. method according to claim 1, it is characterised in that described producing and ethanol material includes to be the yeast cells of ethanol by wood-sugar fermentation.

10. method according to claim 2, it is characterised in that described producing and ethanol substance source home-brewed wine yeast.

11. method according to claim 1, it is characterised in that use xylose as the carbon source for the growth of described producing and ethanol material the second cell mass.

12. method according to claim 1, it is characterised in that described producing and ethanol material the second cell mass includes the inoculum that can be supplied to fermentation system for the described producing and ethanol material of pentose fermentation.

13. method according to claim 1, it is characterised in that farther include to provide the step of additive to described culture medium; Wherein said additive includes at least one nutrient.

14. method according to claim 1, it is characterised in that described producing and ethanol material includes can by the bacterial strain of saccharomyces cerevisiae that xylose is ethanol.

15. method according to claim 1, it is characterised in that the step of described maintenance culture medium, apply described composition including to described culture medium, and described culture medium is maintained at about under 30 degrees Celsius about 120 hours.

16. method according to claim 2, it is characterised in that described producing and ethanol material includes yeast, and described producing and ethanol material the second cell mass includes about 0.35 to 0.55 gram of dry yeast of every gram of sugar.

17. method according to claim 11, it is characterised in that use xylose to supply the efficiency that the first cell mass of described producing and ethanol material is grown to described second cell mass to be at least 80%.

18. method according to claim 13, it is characterised in that described additive includes at least one in yeast extract, carbamide, Diammonium phosphate (DAP), magnesium sulfate or zinc sulfate.

19. method according to claim 7, it is characterised in that xylose is the composition as described biomass and obtains, and supplies to described culture medium with the speed of 0.12 gram of xylose at least per minute.

20. method according to claim 2, it is characterised in that to concentration is culture medium weight at least the 1% of the xylose that described culture medium is supplied.

21. method according to claim 2, it is characterised in that to concentration is culture medium weight at least the 3% of the xylose that described culture medium is supplied.

22. method according to claim 2, it is characterised in that to concentration is culture medium weight about the 6% of the xylose that described culture medium is supplied.

23. method according to claim 1, it is characterised in that the step of described maintenance culture medium, ventilate including to yeast-inoculated body.

24. method according to claim 3, it is characterised in that farther include to supply the step of every volume medium at least 1.0 volume gas streams per minute to described culture medium.

25. method according to claim 19, it is characterised in that described composition includes the xylose of about 2 to 5 weight %.

26. method according to claim 1, it is characterised in that described producing and ethanol material is organism.

27. method according to claim 1, it is characterised in that described biomass include hemicellulose, and produce described tunning by xylose-fermenting.

28. method according to claim 1, it is characterised in that described biomass include cellulose, and produce described tunning by glucose fermentation.

29. method according to claim 19, it is characterised in that described composition includes xylose and glucose, and described producing and ethanol material includes being ethanol by wood-sugar fermentation and being the organism of ethanol by glucose fermentation.

30. method according to claim 29, it is characterised in that described producing and ethanol material includes saccharomyces cerevisiae.

31. for the method breeding producing and ethanol material, described producing and ethanol material, for producing tunning from biomass, said method comprising the steps of:

Culture medium for breeding producing and ethanol material is provided;

Additive is provided to described culture medium;

The composition taking from described biomass is provided, as the carbon source of described producing and ethanol material to described culture medium;

Keep the pH of culture medium of the first cell mass comprising described producing and ethanol material between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that the second cell mass that the first cell mass breeding of described producing and ethanol material is described producing and ethanol material;

Wherein, the second cell mass of described producing and ethanol material is more than the first cell mass of described producing and ethanol material;

Wherein, described biomass include cellulosic material;

Wherein, described cellulosic material includes at least one in corncob, corn plant skin, corn plant leaf and corn plant bar;

Wherein, described composition obtains from described cellulosic material;

Wherein, described composition includes pentose;

Wherein, described pentose includes the carbon source for described producing and ethanol material; With

Wherein said producing and ethanol material includes to be the yeast cells of ethanol by pentose fermentation.

32. method according to claim 31, it is characterised in that the second cell mass of described producing and ethanol material includes the inoculum that can be supplied to fermentation system for the described producing and ethanol material of pentose fermentation.

33. method according to claim 31, it is characterised in that described pentose includes xylose.

34. method according to claim 31, it is characterised in that described producing and ethanol material includes can by the bacterial strain of saccharomyces cerevisiae that xylose is ethanol.

35. method according to claim 31, it is characterised in that the step of described maintenance culture medium, provide described composition including to described culture medium, and described culture medium is maintained at about under 30 degrees Celsius about 120 hours.

36. method according to claim 35, it is characterised in that described culture medium includes sugar, described producing and ethanol material includes yeast, and the second cell mass of described producing and ethanol material includes about 0.35 to 0.55 gram of dry yeast of every gram of sugar.

37. method according to claim 31, it is characterised in that described additive includes at least one in yeast extract, carbamide, Diammonium phosphate (DAP), magnesium sulfate or zinc sulfate.

38. method according to claim 31, it is characterised in that the described step supplying described composition to culture medium, supply xylose including the speed with 0.12 gram of xylose at least per minute to described culture medium.

39. method according to claim 31, it is characterised in that include the described culture medium ventilation comprising described producing and ethanol material.

40. the method according to claim 39, it is characterised in that the described culture medium comprising described producing and ethanol material is ventilated, including the step supplying every volume medium at least 1.0 volume gas streams per minute to described culture medium.

41. method according to claim 33, it is characterised in that described composition includes the xylose of about 2 to 5 weight %.

42. method according to claim 33, it is characterised in that described biomass include hemicellulose, and produce described tunning by xylose-fermenting.

43. method according to claim 33, it is characterised in that described biomass include cellulose, and produce described tunning by glucose fermentation.

44. method according to claim 31, it is characterised in that described composition includes xylose and glucose, and described producing and ethanol material includes being ethanol by wood-sugar fermentation and being the organism of ethanol by glucose fermentation.

45. the method described in claim 44, it is characterised in that described producing and ethanol material includes saccharomyces cerevisiae.

46. method according to claim 31, it is characterised in that further include steps of

The second culture medium for breeding producing and ethanol material is provided;

The second cell mass of described producing and ethanol material is supplied to described second culture medium;

Keep the pH of culture medium of the first cell mass comprising described producing and ethanol material between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that the 3rd cell mass that the second cell mass breeding of described producing and ethanol material is described producing and ethanol material;

Wherein, the cell mass of the 3rd cell mass of described producing and ethanol material is more than the cell mass of the second cell mass of described producing and ethanol material.

47. the method described in claim 46, it is characterized in that, second cell mass that first cell mass breeding is described producing and ethanol material of described producing and ethanol material and the 3rd cell mass that the second cell mass breeding is described producing and ethanol material of described producing and ethanol material, including the total time less than about 80 hours.

48. method according to claim 2, it is characterised in that described offer, for breeding the step of the first culture medium of described producing and ethanol material, including the first stage, and further includes steps of

Put forward the second culture medium breeding producing and ethanol material for use in second stage;

49. method according to claim 48, it is characterised in that provide xylose to described second culture medium, as the carbon source supplying described producing and ethanol substance migration in described second stage.

50. method according to claim 48, it is characterised in that at least big than described first cell mass 200 times of described second cell mass.

51. method according to claim 48, it is characterised in that at least big than described second cell mass 20 times of described 3rd cell mass.

52. method according to claim 50, it is characterized in that, second cell mass that first cell mass breeding is described producing and ethanol material of described producing and ethanol material and the 3rd cell mass that the second cell mass breeding is described producing and ethanol material of described producing and ethanol material, including the total time less than about 80 hours.

53. for the system breeding producing and ethanol material, described producing and ethanol material for producing tunning from biomass in fermentation system, and the described system for breeding producing and ethanol material includes:

First stage, including for keeping the first container comprising the culture medium of described producing and ethanol material;

Second stage, including for keeping the second container of culture medium provided by the described first stage;

Xylose source, this xylose source is provided to described culture medium as producing and ethanol material carbon source in the described first stage;

Xylose source, this xylose source is provided to described culture medium as producing and ethanol material carbon source in described second stage;

Wherein, when being provided to the described first stage, described producing and ethanol material has the first cell mass; When being supplied to described second stage from the described first stage, described producing and ethanol material has the second cell mass; And when providing from described second stage, described producing and ethanol material has the 3rd cell mass;

Wherein, keep the pH of described culture medium in described first container between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that described producing and ethanol material can be bred as described second cell mass;

Wherein, keep the pH of described culture medium in described second container between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that described producing and ethanol material can be bred as described 3rd cell mass;

Wherein, described second cell mass is than big at least 200 times of described first cell mass;

Wherein, described 3rd cell mass is than big at least 20 times of described second cell mass.

54. system according to claim 53, it is characterised in that farther include: for being the equipment of liquid component by described biomass pretreatment, described liquid component comprises the described xylose source for described producing and ethanol substance migration.

55. for the method breeding producing and ethanol material, described producing and ethanol material is for producing tunning from biomass, and described method includes:

Culture medium for breeding producing and ethanol material is provided;

Producing and ethanol material first cell mass with the first cell mass is provided to described culture medium;

The xylose as producing and ethanol material carbon source is supplied to described culture medium, under aerobic conditions, keep the pH of the first cell mass of described producing and ethanol material between about 5.0 to 6.0 and temperature is between about 26 to about 37 degrees Celsius, so that the second cell mass that the first cell mass breeding of described producing and ethanol material is described producing and ethanol material;

And wherein, the second cell mass of described producing and ethanol material is more than the first cell mass of described producing and ethanol material.

56. method according to claim 55, it is characterised in that xylose is available to the sole carbon source of described culture medium.

57. method according to claim 55, it is characterized in that, the described step keeping described culture medium under aerobic conditions, supplies every volume medium at least 1.0 volume gas streams per minute including to described culture medium, and thus described xylose is for Growth of Cells and nonproductive ethanol.

58. method according to claim 55, it is characterised in that to concentration is culture medium weight at least the 3% of the xylose that described culture medium is supplied.

59. method according to claim 55, it is characterised in that to concentration is culture medium weight about the 6% of the xylose that described culture medium is supplied.

60. method according to claim 55, it is characterised in that described biomass include cellulosic material.