CN104357491A - Pretreatment method for producing butanol by fermentation of bagasse - Google Patents

Abstract

The invention discloses a pretreatment method for producing butanol by fermentation of bagasse. The pretreatment method comprises the following steps of mechanical grinding, liquid-state hot-water treatment, microwave treatment, microbial decomposition, ammonia treatment and enzymolysis treatment, after the steps are performed, the decomposition products in each step can be mixed and subjected to step-by-step saccharification and fermentation and simultaneous saccharification and fermentation. By the pretreatment method, the contents of major inhibitors furancarboxaldehyde and hydroxymethyl furfural can be controlled below the concentration without affecting the growth of fermentative bacteria and other inhibitors such as glucuronic acid, p-coumaric acid, syringic acid and ferulic acid are not basically produced. Since the detoxification purification treatment of the pretreated decomposition products is not needed and the pretreated decomposition products are directly subjected to biological fermentation to produce butanol, the pretreatment method has the advantages that the production of inhibitors can be controlled, the cost is low, the yield is high and the environment friendliness is achieved.

Description

A kind of pretreatment process utilizing bagasse fermentation to produce butanols

Technical field

The present invention relates to biomass preprocessing technical field, be specifically related to a kind of pretreatment process utilizing bagasse fermentation to produce butanols.

Background technology

At present, the pretreatment process being directed to filamentary material and half filamentary material of existing multiple maturation obtains sugar with hydrolysis, and the sugar of gained may be used for fermentation to produce biological fuel.

Recently, these methods obtained some progress in industrial biotechnology field, for providing potential chance by the cost-effective agricultural industry waste material that utilizes.Such as, bagasse is a kind of comprehensive material, and be the Main By product in Sugarcane Industry, be also one of Main Agricultural waste material, all richness originates in southern china every year.Bagasse contain the Mierocrystalline cellulose of about 50%, the hemicellulose of 25% and 25% xylogen, with its produce biological butanol be a kind of Perfected process producing biofuel because butanols is compared to traditional ethanol, there is higher calorific value and lower water absorbability.

But, whether feasiblely utilize bagasse to produce butanols, depend on and comprise the many factors such as pre-treatment and detoxification purifying.And traditional pretreatment process, as acidolysis, the normal toxicant producing the growth of serious suppression clostridium.

Much research reports the multiple pretreatment process that can be used for the cereal residue of fibre-rich element and hemicellulose, comprises steam explosion or dilute acid pretreatment method.Acid hydrolyzation is a kind of commonly using and effective pretreatment process, and conventional is especially dilute sulphuric acid.The method is not only simple, and with low cost.But when acidolysis, normal generation one quasi-microorganism inhibitor, this inhibitor can suppress microbial growth when fermenting.Furtural and hydroxymethylfurfural are the major inhibitors produced when biological fermentation.Therefore, before carrying out biological butanol fermentation, first hydrolysate must carry out detoxification purifying, to remove inhibitor.

But, pretreated detoxification purifying is a complicated process, although multiple detoxification purification process obtains research, as lime treatment, method of evaporation, ion-exchange chromatography method, gac and biological treatment, but these methods otherwise detoxification purification effect bad, or cost is too high.At present, main current method utilizes chemical method to remove inhibitor, because chemical method is comparatively simple, cost is also lower.But, when utilizing chemical method, due to the otherness of the chemical structure of inhibitor, cause this method cannot remove all inhibitor.In addition, chemical method also can produce many salts, cause the salt containing high density in fermented liquid, and high salt concentration can produce serious inhibition to fermentable, thus cause the minimizing of biological butanol output.

Therefore, suitable pretreatment process is selected just to seem very important.When selecting pretreatment process, following 2 points need be considered: 1, can the most effectively destroy fibrous texture, to produce sufficient sugar, and prevent the loss to sugar, the generation of inhibitor can also be avoided or reduce simultaneously; 2, pretreatment process needs with low cost, simple.

In the prior art, the pretreatment process can avoided or reduce all major inhibitors and produce also is not had.Therefore, urgently develop one both economically feasibles, all major inhibitors content can be controlled again, produce the pretreatment process of butanols to carry out fermenting to bagasse.

Summary of the invention

The object of the present invention is to provide a kind of pretreatment process utilizing bagasse fermentation to produce butanols, this pretreatment process can control the generation of the toxin suppressing microorganism growth.By controlling the generation of toxin, making bagasse after pretreatment without the need to follow-up purifying detoxification treatment, just can carry out fermentative production butanols, greatly reduce production cost.

This pretreatment process comprises the steps:

1) mechanical disintegration: carrying out segment to length to sugarcane is 5-10cm, dries 2 days, then pulverizes at 65 ± 2 DEG C, crosses 20 mesh sieves, obtains the bagasse powder that particle diameter is less than 830 μm;

2) step 1) gains are soaked 12h, preheat 45s afterwards with steam by liquid heat water treatment: be 250g/L by solid-liquid mass volume ratio under normal temperature in water, then liquid hot water is added, at 200 ± 3 DEG C, reaction 1h, filter after reaction, collect filtrate I and solid residue I;

3) microwave treatment: set-up procedure 2) humidity of gained solid residue I is 40%, and be placed in reactor, then pass into nitrogen and get rid of oxygen, at 4 DEG C, place 8-12h, afterwards reactor is placed in microwave-heating stove, at 120 DEG C, react 7 min, again reaction gains are cooled to normal temperature, with 90% ethanol, reaction gains are embathed, filter afterwards, collect filtrate II and solid residue II;

4) microbial decomposition: baking step 3) gained solid residue II, add nutrient solution afterwards to mix, sterilizing is carried out again at 121 DEG C, after sterilizing, adjusting its humidity is 30%, by percentage to the quality, add the decomposer bacterium liquid of 10% again, in 30 DEG C of bottom fermentation 10 d, filter afterwards, collect filtrate III and solid residue III;

The formula of described nutrient solution is: 30 mg H ₃bO ₃, 20 mg MnCl2.4H2O, 185 mg ZnSO47H ₂o, 20 mg Na ₂moO ₄2H ₂o, 280 mg FeSO ₄h ₂o, 200 mg CuSO ₄, 5 g NH ₄nO ₃, 1 g KH ₂pO ₄, 0.15 g MgSO ₄h ₂o, 0.11 g CaCl ₂with 1 L ddH ₂o, pH are 7.0;

Described decomposer is aspergillus nigeraTCC 1015, trichoderma reeseiaTCC 26921 He penicillium janthinellumone in ATCC 44750;

5) ammonia treatment: use ddH ₂o cleans step 4) gained solid residue III, is dried afterwards, with solid-liquid mass ratio for 1:2, solid residue III after drying is mixed with ammoniacal liquor, under 0.1MPa pressure, in 90 DEG C of reaction 30 min, filter afterwards, collect filtrate IV and solid residue IV;

6) enzymolysis processing: described solid residue IV is soaked 2 h in water, pH is regulated to be 7.0, dried after concentrated, then prepare enzymolysis mixture, this enzymolysis mixture of every 500 ml is made up of following substances: described solid residue IV, 10g cellulase, 100 ml OPTMASH BG, 5 mg paraxin and 2.5 mg kantlex that 50 g have been dried; The pH of adjustment enzymolysis mixture is 5.0, and be 55 DEG C in temperature, shaking speed is under 220rpm, reacts 60 h, filters afterwards, collects filtrate V and solid residue V;

7) filtrate I, filtrate II, filtrate III, filtrate IV and solid residue IV are merged, wait until SSF fermentation, or described filtrate I, filtrate II, filtrate III, filtrate IV and filtrate V are merged and concentrated by rotary evaporation, wait until SHF fermentation.

Preferably, when carrying out microbial decomposition, decomposer used is trichoderma reesei ATCC 26921.

In pre existing processing mode, when being used alone a certain pretreatment mode, Bagasse-cellulose of can not fully degrading, sugar yield is extremely low; As with the coupling of diluted acid method, although sugar yield makes moderate progress, can to reactor produce corrosion and there is pollution problem, meanwhile, also undesirable to the control effects of inhibitor concentration.

First the present invention selects high hot-water cure, first by soaking 12h at normal temperatures in water, preheating 45s afterwards, then add liquid hot water with steam, and at 200 ± 3 DEG C, reaction 1h, just can realize destroying between Mierocrystalline cellulose and xylogen combining firmly molecular structure.This is because first must destroy the structure of Bagasse-cellulose, bagasse is made to be in " loose " state, to be for further processing.At comparatively high pressure, and under the not too high pretreatment condition of temperature, steam-energy fully contacts with the surface of bagasse, thus destroys between Mierocrystalline cellulose and xylogen and combine firmly molecular structure.

Although through the first step pre-treatment, the structure of bagasse presents " loose " state to a certain degree, and bagasse is not still degraded fully, and therefore second step adopts Microwave Pretreatment.At 120 DEG C, to process under microwave through the pretreated bagasse of the first step, the bagasse being in " loose " state can be made further to decompose, but after Microwave Pretreatment, some materials such as bio oil, diesel oil may be produced, by the washing with alcohol of 90%, can wash-out produce oily substance, be convenient to next step microorganism decomposition growth.

After first two steps pre-treatment, bagasse can carry out the 3rd step and decompose, i.e. microbial decomposition.Microorganism mainly utilizes the enzyme system decomposition of cellulose self produced, and when enzyme cuts, Mierocrystalline cellulose is further decomposed.Available decomposer is aspergillus nigeraTCC 1015, trichoderma reeseiaTCC 26921 He penicillium janthinellumone in ATCC 44750, wherein trichoderma reeseithe discomposing effect of ATCC 26921 is best, and this may be because this bacterium is more partial to and utilizes the decomposition of bagasse.

But above a few step is mainly decomposability Mierocrystalline cellulose and hemicellulose, but some xylogen may also not decomposed fully, therefore, 4th step adopts ammoniacal liquor to soak, the object that ammoniacal liquor soaks is that the Mierocrystalline cellulose making to be destroyed is separated with xylogen, only have and be separated xylogen, the enzymolysis in next step just can make Mierocrystalline cellulose obtain further decomposing again.In ammoniacal liquor soaks, soak time, pressure and temperature outbalance, by suitable setting, can improve the decomposition efficiency of bagasse, and reduce the generation of inhibitor.After above a few step pre-treatment, final step adopts enzymolysis.

The order of above-mentioned pre-treatment step is fixing.By above-mentioned steps process, not only can obtain higher sugar yield, and in hydrolyzed solution, all main inhibitor contents, lower than the concentration affecting zymophyte growth, or are low to moderate inspection and do not measure.

Size due to bagasse affects the complexity of decomposition, and generally speaking, less bagasse more easily decomposes, but considers Energy Consumption Factors, and the size of bagasse controlled below 860 μm, not only discomposing effect is good, and energy consumption is lower.

Liquid heat water law is compared to diluted acid method, and without the need to adding sour reagent, can not cause corrosion to reactor, the residue in the hydrolyzed solution of generation is also less, is a kind of comparatively gentle pretreatment process.But its sugarcane resistates rate of recovery is lower, the object of effectively decomposing bagasse when being used alone, cannot be reached.

After the process of liquid heat water law, recycling microwave processes, can the output of raising reducing sugar by a relatively large margin Reaction time shorten and reduction energy consumption.When carrying out microwave treatment, microwave can accelerate the destruction to crystalline structure, makes that microorganism is easier decomposes further to bagasse.After being decomposed by decomposer, the bagasse rate of decomposition of about 10% can be improved again.

Preferably, when utilizing decomposer to decompose bagasse, utilize trichoderma reesei ATCC 26921better decomposition efficiency can be realized.

When carrying out ammonia treatment, because ammonia is the good solvent of xylogen, xylogen can be separated with lignocellulose, thus subsequent fiber element enzyme can be improved to the decomposition efficiency of bagasse.

After above-mentioned process, recycling enzymolysis processing, the hydrolysis bagasse of maximizing.

Beneficial effect of the present invention:

1, can effectively hydrolyzing bagasse, percent hydrolysis can reach 74.41%;

2, can obtain the glucose of high density, glucose concn can reach 15.21 ± 1.65g/L;

3, pretreatment condition is gentle, and energy consumption is low, not etching apparatus;

4, without the need to following purification steps, fermentable production cost is low;

5, major inhibitors furtural and hydroxymethyl furfural content low, do not affect the growth of organism of fermentation, simultaneously substantially do not produce other inhibitor such as glucuronic acid, P-coumaric acid, syringic acid and forulic acid.

Accompanying drawing explanation

1, Figure 1 shows that the schematic diagram of pretreatment process step of the present invention;

2, shown in Fig. 2, during for utilizing bagasse hydrolysate (filtrate) as fermentation substrate, the graph of a relation of tunning and fermentation time; Wherein HAN is expressed as: adopt in step 4) aspergillus nigerdescribed filtrate I, filtrate II, filtrate III, filtrate IV and filtrate V as zymophyte, and merge by ATCC 1015, ferment as SHF; HTR is expressed as: adopting in step 4) trichoderma reeseidescribed filtrate I, filtrate II, filtrate III, filtrate IV and filtrate V as zymophyte, and merge by ATCC 26921, ferment as SHF; HPJ is expressed as: adopting in step 4) penicillium janthinellumdescribed filtrate I, filtrate II, filtrate III, filtrate IV and filtrate V as zymophyte, and merge by ATCC 44750, ferment as SHF; HANNY is expressed as: adopt in step 4) aspergillus nigerfiltrate I, filtrate II, filtrate III, filtrate IV and solid residue IV as zymophyte, and merge by ATCC 1015, ferment as SSF; HTRNY is expressed as: adopt in step 4) trichoderma reeseifiltrate I, filtrate II, filtrate III, filtrate IV and solid residue IV as zymophyte, and merge by ATCC 26921, ferment as SSF; HPYNY is expressed as: adopt in step 4) penicillium janthinellumfiltrate I, filtrate II, filtrate III, filtrate IV and solid residue IV as zymophyte, and merge and concentrated by rotary evaporation by ATCC 44750, ferment as SSF;

3, shown in Fig. 3, during for utilizing bagasse hydrolysate (filtrate) as fermentation substrate, the variation diagram of pH in SSF and SHF fermenting process; Wherein, the implication of HAN, HTR, HPJ, HANNY, HTRNY, HPJNY and consistent in Fig. 2; A is SSF fermentation; B is SHF fermentation;

4, shown in Fig. 4, in SSF and SHF fermenting process, to the conversion service condition of different sugar composition; Wherein a, b, c are SHF fermentation, and b, d, f are SSF fermentation, the implication of HAN, HTR, HPJ, HANNY, HTRNY, HPJNY and consistent in Fig. 2;

5, shown in Fig. 5, at fermentation termination, the total resistates carbohydrate produced by different fermentations substrate; Wherein, the implication of HAN, HTR, HPJ, HANNY, HTRNY, HPJNY and consistent in Fig. 2;

6, shown in Fig. 6, during for utilizing pure glucose as fermentation substrate, during the fermentation, the variation diagram of different parameters;

7, shown in Fig. 7, during for utilizing sugar composition as fermentation substrate, the variation diagram of different product, sugar and pH.

Embodiment

Below by embodiment, the present invention is specifically described; what be necessary to herein means out is that following examples are just for being further detailed the present invention; limiting the scope of the invention can not be interpreted as; some nonessential improvement and adjustment that the person skilled in the art in this field makes according to foregoing invention content, still belong to protection scope of the present invention.

Embodiment 1:

1) 500g sugarcane being carried out segment to length is 5-10cm, dries 2 days, then pulverize at 65 ± 2 DEG C, crosses 20 mesh sieves, obtains the bagasse powder that particle diameter is less than 830 μm;

2) step 1) gains are soaked 12h under normal temperature in the water of 2L, preheat 45s afterwards with steam, then add liquid hot water, at 200 ± 3 DEG C, reaction 1h, filters after reaction, collects filtrate I and solid residue I;

3) set-up procedure 2) humidity of gained solid residue is 40%, and be placed in three neck round-bottomed flasks, then pass into nitrogen and get rid of oxygen, at 4 DEG C, place 12h, afterwards reactor is placed in microwave-heating stove and reacts 7 min at 120 DEG C, discharge hot gas, drop to after room temperature until temperature, with 90% ethanol, reaction gains are embathed, filter afterwards, collect filtrate II and solid residue II;

4) baking step 3) gained solid residue, add nutrient solution afterwards and mix, then carry out sterilizing at 121 DEG C, after sterilizing, the humidity of adjustment mixture is 30%, then adds 20ml aspergillus nigeraTCC 1015 bacterium liquid, in 30 DEG C of bottom fermentation 10 d, filters afterwards, collects filtrate III and solid residue III;

The formula of nutrient solution is: 30 mg H ₃bO ₃, 20 mg MnCl2.4H2O, 185 mg ZnSO47H ₂o, 20 mg Na ₂moO ₄2H ₂o, 280 mg FeSO ₄h ₂o, 200 mg CuSO ₄, 5 g NH ₄nO ₃, 1 g KH ₂pO ₄, 0.15 g MgSO ₄h ₂o, 0.11 g CaCl ₂and 1 L ddH ₂o, pH are 7.0;

5) ddH is used ₂o cleans step 4) gained solid residue, is dried afterwards, with solid-liquid mass ratio for 1:2, solid residue III after drying is mixed with ammoniacal liquor, under high pressure (0.1MPa), in 90 DEG C of reaction 30 min, filter afterwards, collect filtrate IV and solid residue IV;

6) step 5) gained solid residue IV is soaked 2 h in the distilled water of 900ml, with the H of 1% ₂sO ₄regulate pH to be 7.0, with Rotary Evaporators, it is concentrated, then dried at 60 DEG C.Then prepare enzymolysis mixture, this enzymolysis mixture of every 500 ml is made up of following substances: solid residue IV, 10g cellulase, 100 ml OPTMASH BG, 5 mg paraxin and 2.5 mg kantlex that 50 g have been dried; The pH of adjustment enzymolysis mixture is 5.0, and be 55 DEG C in temperature, shaking speed is under 220rpm, reacts 60 h, filters afterwards, collects filtrate V and solid residue V.

The solid recovery rate of each step is in table 1.

Embodiment 2

1) 500g sugarcane being carried out segment to length is 5-10cm, dries 2 days, then pulverize at 65 ± 2 DEG C, crosses 20 mesh sieves, obtains the bagasse powder that particle diameter is less than 830 μm;

2) step 1) gains are soaked 12h under normal temperature in the water of 2L, preheat 45s afterwards with steam, then add liquid hot water, at 200 ± 3 DEG C, reaction 1h, filters after reaction, collects filtrate I and solid residue I;

3) set-up procedure 2) humidity of gained solid residue is 40%, and be placed in three neck round-bottomed flasks, then pass into nitrogen and get rid of oxygen, at 4 DEG C, place 8h, afterwards reactor is placed in microwave-heating stove and reacts 7 min at 120 DEG C, discharge hot gas, drop to after room temperature until temperature, with 90% ethanol, reaction gains are embathed, filter afterwards, collect filtrate II and solid residue II;

4) baking step 3) gained solid residue, add nutrient solution afterwards and mix, then carry out sterilizing at 121 DEG C, after sterilizing, the humidity of adjustment mixture is 30%, then adds 20ml aspergillus nigeraTCC 1015 bacterium liquid, in 30 DEG C of bottom fermentation 10 d, filters afterwards, collects filtrate III and solid residue III;

The formula of nutrient solution is: 30 mg H ₃bO ₃, 20 mg MnCl2.4H2O, 185 mg ZnSO47H ₂o, 20 mg Na ₂moO ₄2H ₂o, 280 mg FeSO ₄h ₂o, 200 mg CuSO ₄, 5 g NH ₄nO ₃, 1 g KH ₂pO ₄, 0.15 g MgSO ₄h ₂o, 0.11 g CaCl ₂and 1 L ddH ₂o, pH are 7.0;

5) ddH is used ₂o cleans step 4) gained solid residue, is dried afterwards, with solid-liquid mass ratio for 1:2, solid residue III after drying is mixed with ammoniacal liquor, under high pressure (0.1MPa), in 90 DEG C of reaction 30 min, filter afterwards, collect filtrate IV and solid residue IV;

6) step 5) gained solid residue IV is soaked 2 h in the distilled water of 900ml, with the H of 1% ₂sO ₄regulate pH to be 7.0, with Rotary Evaporators, it is concentrated, then dried at 60 DEG C.Then prepare enzymolysis mixture, this enzymolysis mixture of every 500 ml is made up of following substances: solid residue IV, 10g cellulase, 100 ml OPTMASH BG, 5 mg paraxin and 2.5 mg kantlex that 50 g have been dried; The pH of adjustment enzymolysis mixture is 5.0, and be 55 DEG C in temperature, shaking speed is under 220rpm, reacts 60 h, filters afterwards, collects filtrate V and solid residue V.

The solid recovery rate of each step is in table 2.

Embodiment 3

1) 500g sugarcane being carried out segment to length is 5-10cm, dries 2 days, then pulverize at 65 ± 2 DEG C, crosses 20 mesh sieves, obtains the bagasse powder that particle diameter is less than 830 μm;

2) step 1) gains are soaked 12h under normal temperature in the water of 2L, preheat 45s afterwards with steam, then add liquid hot water, at 200 ± 3 DEG C, reaction 1h, filters after reaction, collects filtrate I and solid residue I;

3) set-up procedure 2) humidity of gained solid residue is 40%, and be placed in three neck round-bottomed flasks, then pass into nitrogen and get rid of oxygen, at 4 DEG C, place 10h, afterwards reactor is placed in microwave-heating stove and reacts 7 min at 120 DEG C, discharge hot gas, drop to after room temperature until temperature, with 90% ethanol, reaction gains are embathed, filter afterwards, collect filtrate II and solid residue II;

4) baking step 3) gained solid residue, add nutrient solution afterwards and mix, then carry out sterilizing at 121 DEG C, after sterilizing, the humidity of adjustment mixture is 30%, then adds 20ml penicillium janthinellumaTCC 44750 bacterium liquid, in 30 DEG C of bottom fermentation 10 d, filters afterwards, collects filtrate III and solid residue III;

The formula of nutrient solution is: 30 mg H ₃bO ₃, 20 mg MnCl2.4H2O, 185 mg ZnSO47H ₂o, 20 mg Na ₂moO ₄2H ₂o, 280 mg FeSO ₄h ₂o, 200 mg CuSO ₄, 5 g NH ₄nO ₃, 1 g KH ₂pO ₄, 0.15 g MgSO ₄h ₂o, 0.11 g CaCl ₂and 1 L ddH ₂o, pH are 7.0;

5) ddH is used ₂o cleans step 4) gained solid residue, is dried afterwards, with solid-liquid mass ratio for 1:2, solid residue III after drying is mixed with ammoniacal liquor, under high pressure (0.1MPa), in 90 DEG C of reaction 30 min, filter afterwards, collect filtrate IV and solid residue IV;

6) step 5) gained solid residue IV is soaked 2 h in the distilled water of 900ml, with the H of 1% ₂sO ₄regulate pH to be 7.0, with Rotary Evaporators, it is concentrated, then dried at 60 DEG C.Then prepare enzymolysis mixture, this enzymolysis mixture of every 500 ml is made up of following substances: solid residue IV, 10g cellulase, 100 ml OPTMASH BG, 5 mg paraxin and 2.5 mg kantlex that 50 g have been dried; The pH of adjustment enzymolysis mixture is 5.0, and be 55 DEG C in temperature, shaking speed is under 220rpm, reacts 60 h, filters afterwards, collects filtrate V and solid residue V.

The solid recovery rate of each step is in table 3.

Embodiment 4

1) substep diastatic fermentation method: respectively the filtrate I in the middle of embodiment 1, embodiment 2 and embodiment 3, filtrate II, filtrate III, filtrate IV and filtrate V are merged and concentrates.In each embodiment, the sugared composition of the filtrate after merging is as shown in table 4.

Filtrate after being merged by 150ml is placed in the anaerobic bottle of 250ml specification, and seals with butyl rubber plug; Add 0.75g yeast extract and 1.5g peptone, the NaOH with 1% regulates pH to 6.5, after 115 DEG C of sterilizing 20 min, is cooled to normal temperature; Add again after 0.5 ml sterilizing mixing solutions, this mixing solutions contains 50g/L KH ₂pO ₄, 50g/L K ₂hPO ₄, 220g/L CH3COONH4,0.1g/L para-amino benzoic acid, 0.1g/L vitamins B, 0.001g/L vitamin H, 20 g/L MgSO ₄7H ₂o, 1 g/L MnSO ₄h ₂o, 1 g/L FeSO ₄7H ₂o and 1 g/L NaCl; Then ferment to OD600=1.5.

2) simultaneous saccharification and fermentation method: the filtrate I in the middle of embodiment 1, embodiment 2 and embodiment 3, filtrate II, filtrate III and filtrate IV are merged respectively, regulates pH to 7.0.In each embodiment, the composition of the filtrate after merging is as shown in table 5.

Fermentation step and enzymolysis carry out simultaneously, and added trace element is consistent with distribution diastatic fermentation method with fermentation condition.Leavening temperature is 37 DEG C, and fermentation time is 96h, during the fermentation, every for some time, samples through rubber plug with asepsis injector, and institute's sample thief is at 4 DEG C, and centrifugal 20min under 8000rpm, then carries out the detection of sugar, organic acid, pH and ABE production.

Embodiment 5

The detection of inhibitor content: at DIONEX UltiMate 3000 liquid chromatograph, utilizes Aminex HPX-87H chromatographic column to detect butyric acid, acetic acid, glucuronic acid, P-coumaric acid, syringic acid and ferulaic acid content; The mensuration of furtural and 5 hydroxymethyl furfural is with reference to " Determination of 5-Hydroxymethylfurfural in Vinegar Samples by HPLC " (Theobald A, M ü ller A, Anklam E. J. Agric. Food. Chem. 1998,46 (5): 1850-1854.) in method.

After all pre-treatment step, in embodiment 1-3, each inhibitor content is in table 6.

Comparative example

Under fermentation condition identical with embodiment 4, pure glucose or sugar composition (10g/L glucose, 42g/L wood sugar, 3.5g/L pectinose, 3.5g/L cellobiose, 3g/L semi-lactosi and 1.5g/L seminose) is utilized to substitute pretreated hydrolysate (filtrate).

Embodiment 6

Tunning detects: alcohols material (acetone, butanols, ethanol) utilizes gas chromatograph to detect; The detection of total solids resistates carbohydrate is with reference to " Carbohydrate analysis by a phenol – sulfuric acid method in microplate format " (Masuko T, Minami A, Iwasaki N, Majima T, Nishimura S-I, Lee YC:Anal. Biochem. 2005,339 (1): 69-72.) in method; Content of cellulose utilizes spectrophotometer to detect; The detection of content of lignin is with reference to " Determination of lignin in herbaceous plants by an improved acetyl bromide procedure " (Iiyama K, Wallis AFA J. Sci. Food Agric. 1990,51 (2): 145-161.) in method; Ash oontent detects after drying in 500 DEG C in sintering oven.

In fermenting process, each parameter index of tunning is in table 7.

Claims (2)

1. utilize bagasse to ferment and produce a pretreatment process for butanols, it is characterized in that: described method comprises the steps:

1) mechanical disintegration: carrying out segment to length to sugarcane is 5-10cm, dries 2 days, then pulverizes at 65 ± 2 DEG C, crosses 20 mesh sieves, obtains the bagasse powder that particle diameter is less than 830 μm;

2) step 1) gains are soaked 12h, preheat 45s afterwards with steam by liquid heat water treatment: be 250g/L by solid-liquid mass volume ratio under normal temperature in water, then liquid hot water is added, at 200 ± 3 DEG C, reaction 1h, filter after reaction, collect filtrate I and solid residue I;

3) microwave treatment: set-up procedure 2) humidity of gained solid residue I is 40%, and be placed in reactor, then pass into nitrogen and get rid of oxygen, at 4 DEG C, place 8-12h, afterwards reactor is placed in microwave-heating stove, at 120 DEG C, react 7 min, again reaction gains are cooled to normal temperature, with 90% ethanol, reaction gains are embathed, filter afterwards, collect filtrate II and solid residue II;

4) microbial decomposition: baking step 3) gained solid residue II, add nutrient solution afterwards to mix, sterilizing is carried out again at 121 DEG C, after sterilizing, adjusting its humidity is 30%, by percentage to the quality, add the decomposer bacterium liquid of 10% again, in 30 DEG C of bottom fermentation 10 d, filter afterwards, collect filtrate III and solid residue III;

The formula of described nutrient solution is: 30 mg H ₃bO ₃, 20 mg MnCl2.4H2O, 185 mg ZnSO47H ₂o, 20 mg Na ₂moO ₄2H ₂o, 280 mg FeSO ₄h ₂o, 200 mg CuSO ₄, 5 g NH ₄nO ₃, 1 g KH ₂pO ₄, 0.15 g MgSO ₄h ₂o, 0.11 g CaCl ₂with 1 L ddH ₂o, pH are 7.0;

Described decomposer is aspergillus nigeraTCC 1015, trichoderma reeseiaTCC 26921 He penicillium janthinellumone in ATCC 44750;

5) ammonia treatment: use ddH ₂o cleans step 4) gained solid residue III, is dried afterwards, with solid-liquid mass ratio for 1:2, solid residue III after drying is mixed with ammoniacal liquor, under 0.1MPa pressure, in 90 DEG C of reaction 30 min, filter afterwards, collect filtrate IV and solid residue IV;

6) enzymolysis processing: described solid residue IV is soaked 2 h in water, pH is regulated to be 7.0, dried after concentrated, then prepare enzymolysis mixture, this enzymolysis mixture of every 500 ml is made up of following substances: described solid residue IV, 10g cellulase, 100 ml OPTMASH BG, 5 mg paraxin and 2.5 mg kantlex that 50 g have been dried; The pH of adjustment enzymolysis mixture is 5.0, and be 55 DEG C in temperature, shaking speed is under 220rpm, reacts 60 h, filters afterwards, collects filtrate V and solid residue V;

7) filtrate I, filtrate II, filtrate III, filtrate IV and solid residue IV are merged, wait until SSF fermentation, or described filtrate I, filtrate II, filtrate III, filtrate IV and filtrate V are merged and concentrated by rotary evaporation, wait until SHF fermentation.

2. method according to claim 1, is characterized in that: in described method, and decomposer used in step 4) is trichoderma reesei ATCC 26921.