Apparatus for producing saccharide solution that uses biomass as feedstock

Abstract

An apparatus for treating a biomass feedstock at a high temperature, includes a cooling device 90 for cooling a biomass treated liquid at a high temperature; an enzymatic saccharification tank 103 for saccharifying a cooled treated liquid 101B with an enzyme; a solid-liquid separation apparatus 112 for removing water-slightly soluble substances contained in a saccharide solution 104 taken out from the enzymatic saccharification tank 103 and a foreign substance removing unit 113 provided with a microfiltration (MF) membrane 113a; a dilution tank 132, disposed downstream of the foreign substance removing unit 113, for diluting the saccharide solution from which the water-slightly soluble substances are removed by adding water thereto; a water separation unit 116, provided with a reverse osmosis (RO) membrane 116a, for removing water 114 from the diluted saccharide solution so as to obtain concentrated saccharide solution 115.

Claims

1. An apparatus for producing saccharide solution that uses biomass as a feedstock comprising: a hydrothermal decomposition apparatus that brings a biomass feedstock including at least cellulose, hemicellulose, and lignin into countercurrent contact with pressurized hot water and performs a high temperature and high pressure treatment in a temperature range of 180 C. to 240 C. so as to produce a biomass treated liquid containing a pressurized hot water soluble component of the biomass feedstock; a cooling means that is provided downstream of the hydrothermal decomposition apparatus and cools the biomass treated liquid which has been discharged from the hydrothermal decomposition apparatus; an enzymatic saccharification tank that is provided downstream of the cooling means and saccharifies the cooled treated liquid with an enzyme so as to produce a saccharide solution; a saccharide solution purification means that is provided downstream of the enzymatic saccharification tank and removes a water-slightly soluble substance from the saccharide solution, the water-slightly soluble substance being a substance that is contained in the pressurized hot water soluble component, and solidified or colloidal due to the cooling; a dilution tank that is disposed downstream of the saccharide solution purification means and adds water thereto so as to dilute the saccharide solution from which the water-slightly soluble substance has been removed; a water separation unit that is provided with a reverse osmosis (RO) membrane and removes water from the diluted saccharide solution so as to obtain concentrated saccharide solution; a first measurement unit that measures both of a turbidity and a light absorbance of the saccharide solution from which the water-slightly soluble substance has been removed, wherein the saccharide solution purification means includes: a saccharide solution purification tank that is provided downstream of the enzymatic saccharification tank to receive and retain the saccharide solution taken from the enzymatic saccharification tank; a first circulation line that connects between a bottom portion of the saccharide solution purification tank and an upper portion of the saccharide solution purification tank to circulate the saccharide solution; a second circulation line that connects between the bottom portion of the saccharide solution purification tank and the upper portion of the saccharide solution purification tank to circulate the saccharide solution; a solid-liquid separation apparatus that is interposed in the first circulation line and removes the water-slightly soluble substance contained in the saccharide solution circulating in the first circulation line; a foreign substance removing unit that is interposed in the second circulation line and has a microfiltration (MF) membrane by which the water-slightly soluble substance circulating in the second circulation line is further removed and through which the saccharide solution from which the remaining water-slightly soluble substance is thus removed is passed.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 is a schematic view of an apparatus for producing saccharide solution that uses biomass as a feedstock according to Example 1.

(2) FIG. 2 is a schematic view of a fermentation apparatus that uses biomass as a feedstock according to Example 2.

(3) FIG. 3 is a schematic view of a fermentation apparatus for producing saccharide solution that uses biomass as a feedstock according to Example 3.

(4) FIG. 4 is a schematic view of a fermentation apparatus for producing saccharide solution that uses biomass as a feedstock according to Example 4.

(5) FIG. 5 is a measured chart of a high-performance liquid chromatography.

(6) FIG. 6 is a view illustrating results of decreased amounts of carbon dioxide in an ethanol fermentation test.

(7) FIG. 7 is a view illustrating results of decreased amount of carbon dioxide in an ethanol fermentation test.

(8) FIG. 8 is a schematic view illustrating a hydrothermal decomposition apparatus of biomass according to Example 1.

DESCRIPTION OF EMBODIMENTS

(9) The present invention will be explained in detailed detail in reference to views below. The present invention is not limited to these Examples. In addition, constituent features described in Examples below include one which can be easily considered by those skilled in the art and ones which are substantially the same as those.

Example 1

(10) A fermentation apparatus using a biomass feedstock according to the present invention will be explained in reference to views.

(11) FIG. 8 is the schematic view illustrating a hydrothermal decomposition apparatus of biomass according to Example 1.

(12) First, a hydrothermal decomposition apparatus (50) in which a high temperature and high pressure treatment is performed at a temperature range of 180 C. to 240 C. while a biomass feedstock including at least cellulose, hemicellulose and lignin is brought into countercurrent contact with pressurized hot water is explained using FIG. 8.

(13) The hydrothermal decomposition apparatus 50 is a vertical type apparatus, as illustrated in FIG. 3, but the present invention is not limited thereto, and may be an inclined type or horizontal-type hydrothermal decomposition apparatus. As the hydrothermal decomposition apparatus, the inclined type and the vertical type are more preferable. This is because gases generated during hydrothermal decomposition reaction and gases carried in a feedstock can quickly escape from upper parts, which is preferable. In addition, because decomposition products are extracted with pressurized hot water 55, a concentration of extracts is increased from an upper side toward a bottom side, which is thus preferable in terms of extraction efficiency.

(14) According to the hydrothermal decomposition apparatus 50 of the present Example, a conveying screw means 54 is provided; as a result, 1) a solid component can be conveyed by solid-liquid counter-flow, 2) solid-liquid separation can be performed in an apparatus body 53, and 3) mixing of pressurized hot water 55 on the surface and the inside of the solid is advanced in the apparatus body 53, whereby the reaction is promoted.

(15) In the hydrothermal decomposition apparatus 50 according to the present Example, a biomass feedstock (for example, a straw, and the like in the present Example) 51 is supplied through a biomass supplying apparatus 52 at a normal pressure or under pressure; pressurized hot water 55 is supplied from an edge side different from the supplying side of the biomass feedstock 51 into the inside of the apparatus body 53; hydrothermal decomposition is performed while a biomass feedstock 11 is brought into countercurrent contact with the pressurized hot water 55 to transfer a lignin component and a hemicellulose component into the pressurized hot water 55, thereby separating the lignin component and the hemicellulose component from the biomass feedstock 51; and a biomass solid component 56 is discharged through a biomass discharging apparatus 57 under pressure or at a normal pressure. In the view, reference sign 58 shows pressurized nitrogen.

(16) Here, the biomass feedstock, which is supplied to the hydrothermal decomposition apparatus 50, is not particularly limited, and refers to an accumulation of organisms or organic substances derived from an organism incorporated in a material cycle in the global biosphere (see JIS K 3600 1258). In the present invention, it is particularly preferable to use cellulose resources such as wood including, for example hardwoods, and herbaceous plants, agricultural wastes, food wastes.

(17) The particle size of the biomass feedstock is not particularly limited, and it is preferable to mill it to a size of, for example, five mm or less.

(18) In the present Example, a pre-treatment using a mill as a pretreatment apparatus may be performed before the biomass is supplied. Washing using a washing apparatus may also be performed.

(19) When as the biomass feedstock was used for, for example, hull, it can be supplied to the hydrothermal decomposition apparatus 50 as it is, without the mill treatment.

(20) It is preferable to set a reaction temperature in the hydrothermal decomposition apparatus 50 at a range of 180 C. to 240 C., more preferably 200 C. to 230 C.

(21) This is because a hydrothermal decomposition speed is slow at a to temperature of lower than 180 C., and thus a long retention time is required, leading to increasing in size of the apparatus, which is not preferable. On the other hand, when the temperature is higher than 240 C., the decomposition speed becomes excessive, which increases the transfer of the cellulose component from the solid to the liquid side and, at the same time, promotes the excessive decomposition of the hemicellulose saccharides, which is not preferable.

(22) The hemicellulose component starts to dissolve at about 140 C., the cellulose starts to dissolve at about 230 C., and the lignin component starts at about 140 C. It is preferable to set a temperature to a range of 180 C. to 240 C., where the cellulose remains on the solid component side, and the hemicellulose component and the lignin component have sufficient decomposition speeds.

(23) It is preferable to set a reaction pressure to a pressure about 0.1 to 0.5 MPa higher than a saturated steam pressure of water having each temperature at which the pressurized hot water 55 exists inside the apparatus body 53.

(24) The reaction time is preferably 20 minutes or less, or from three minutes to 10 minutes. This is because if the reaction is continued for a long time, a ratio of the excessively decomposed products is increased, which is not preferable.

(25) In the hydrothermal decomposition apparatus 50, when the biomass feedstock 51 is brought into countercurrent contact with the pressurized hot water 55, it is preferable to make uniform pressurized hot water flow.

(26) Biomass treated liquid at a high temperature 101A in the present invention can be obtained by using such a hydrothermal decomposition apparatus 50.

(27) The pressurized hot water 55 may have any pH value of alkaline, neutral and acidic ranges. When it is alkaline, sodium hydroxide, hydrated lime, ammonia, and the like may be used, and when it is acidic, dilute sulfuric acid, hydrochloric acid, phosphoric acid, and the like may be used. When the pressurized hot water 55 is alkaline or acidic, it has an advantage in which the pre-treatment of the biomass can be efficiently advanced. On the other hand, however, it has disadvantages in which it is necessary to complete the pH adjustment before enzyme is thrown, and it is necessary to prepare chemicals for adjusting the pH and to provide a pH adjusting apparatus.

(28) FIG. 1 is the schematic view of the apparatus for producing saccharide solution that uses biomass as a feedstock according to Example 1.

(29) Next, an apparatus for producing saccharide solution that uses biomass as feedstock will be explained using FIG. 1, which includes cooling means 90 in which the biomass treated liquid at a high temperature 101A, which has been discharged from the hydrothermal decomposition treatment apparatus 50 in FIG. 8, is cooled; an enzymatic saccharification tank 103 in which a cooled treated liquid 101B is subject to saccharification with an enzyme; a solid-liquid separation apparatus 112 in which the water-slightly soluble substances contained in saccharide solution 104, which has been taken out from the enzymatic saccharification tank 103, are removed and a foreign substance removing unit 113 provided with a microfiltration (MF) membrane 113a; a dilution tank 132 which is disposed downstream of the foreign substance removing unit 113 and in which the saccharide solution from which the water-slightly soluble substances have been removed is diluted by adding water thereto; and a water separation unit 116 provided with a reverse osmosis (RO) membrane 116a capable of removing water 114 from the diluted saccharide solution to obtain concentrated saccharide solution 115.

(30) When the solid-liquid separation apparatus 112 for removing the water-slightly soluble substances and the foreign substance removing unit 113 provided with the microfiltration (MF) membrane 113a are disposed, water-slightly soluble substances 111 are removed from the biomass treated liquid at a high temperature 101A generated by decomposing the biomass feedstock in conditions of a high temperature and a high pressure, whereby the saccharide solution 104 including less impurities can be obtained.

(31) Here, it has been confirmed by the present inventors that the water-slightly soluble substances in the present invention are substances different from furfural and HMF (5-hydroxymethyl furfural), which are generally known as fermentation inhibitory substances, though further details about such substances have not been unraveled yet, as explained in Experiment Examples described below.

(32) For example, according to a known hydrothermal decomposition treatment, the biomass and the pressurized hot water are supplied so that they are brought into countercurrent contact with each other, thereby performing the hydrothermal reaction due to the internal heat exchange, as described above. At that time, the reaction condition is that a temperature is high such that the internal temperature is from 180 C. to 240 C. and, at the same time, a pressure 0.1 to 0.4 MPa higher than the saturated vapor pressure of water having a temperature at that time is applied, and thus reaction decomposition products of lignin and the like are included in the biomass treated liquid at a high temperature (pressurized hot water effluent) after the reaction.

(33) The pressurized hot water soluble component dissolved in this biomass treated liquid at a high temperature is cooled to an enzymatic saccharification temperature (for example, 60 C. or lower), in a saccharification step of a subsequent step, and thus a part of the pressurized hot water soluble component, which has been once dissolved in pressurized hot water, is precipitated, and is solidified or is formed into a colloidal water-slightly soluble substance.

(34) When the biomass feedstock is subjected to a decomposition treatment at an ordinary temperature, accordingly, these water-slightly soluble substances do not exist on the liquid side, and thus it can be assumed that the water-slightly soluble substances are unique substances in the biomass treated liquid at a high temperature.

(35) According to the present invention, such water-slightly soluble substances are removed by means of a saccharide solution purification means 110, whereby the purification is performed in the saccharification treatment, and thus in, by way of example, a fermentation step of subsequent steps, it is possible to perform good fermentation treatment and to improve a fermentation efficiency in an alcohol fermentation, as explained in Experiment Examples described above.

(36) Here, the saccharide solution purification means 110 includes, as illustrated in FIG. 1, the solid-liquid separation apparatus 112 in which solid residue such as lignin including the water-slightly soluble substances 111 is separated from the saccharide solution 104 taken out from the enzymatic saccharification tank 103, and the foreign substance removing unit 113 provided with the microfiltration (MF) membrane 113a in which the remaining water-slightly soluble substance is further removed from the saccharide solution 104, which has been separated in the solid-liquid separation apparatus 112.

(37) As the solid-liquid separation apparatus 112, for example, a screw decanter, a sand separator, an MF membrane, and the like may be used. The solid substances are removed by using this to protect the RO membrane 116a.

(38) An apparatus for producing saccharide solution 100A that uses biomass as feedstock according to the present Example includes the dilution tank 132 in which water (RO water) 131 was added to the saccharide solution 104 from which the water-slightly soluble fermentation inhibitory substances have been removed to dilute it. In FIG. 1, reference sign P.sub.6 shows a liquid sending pump for sending dilution water 133 to the water separation unit 116.

(39) When addition of water before the solid substances are removed, only the water-slightly soluble fermentation inhibitory substance in an amount of its solubility is dissolved, and thus a concentration of the fermentation inhibitory substance is not reduced. When the dilution with water is performed after the water-slightly soluble fermentation inhibitory substances are removed, however, it is possible to remove organic acids (for example, acetic acid) and the like other than the decomposition products such as lignin, and to perform the fermentation with high efficiency in the subsequent steps.

(40) Water 114 was removed from the dilution water 133, which has been diluted in the dilution tank 132, through the water separation unit 116 having the RO (Reverse Osmosis: RO) membrane 116a to concentrate it to a pre-determined concentration, thereby obtaining concentrated saccharide solution 115.

(41) In the water separation unit 116, a loose RO membrane, a nanofiltration membrane (NF membrane), and the like may be used.

(42) In the present Example, in order to remove the solid residue 111 such as lignin including the water-slightly soluble substance, the solid-liquid separation treatment in the solid-liquid separation apparatus 112 and the membrane treatment in the foreign substance removing unit 113 are performed, whereby the water-slightly soluble substances can be effectively removed.

(43) In particular, many zymocytes capable of fermenting saccharide solution (C5 saccharide), which is obtained by subjecting the hemicellulose, which is the pressurized hot water soluble component, to the saccharification, have poor ethanol resistance and poor resistance to fermentation inhibitory substances, and thus it is important to fit to the resistances of the zymocyte. The present invention can realize it. In fermentation in which a recombinant yeast cannot be used, the resistance of the zymocyte remarkably becomes problems, but the problems can be solved by the present invention.

(44) The decomposition products of the water-slightly soluble substance including lignin are removed during the treatment of the saccharide solution 104, and thus the precipitation of the foreign substances on the RO membrane 116a, which is disposed downstream and is used for separating water, can be prevented, whereby the membrane life of the RO membrane 116a can be prolonged.

(45) Next, the treatment procedure of the saccharide solution production apparatus 100A using this biomass treated liquid at a high temperature will be explained.

(46) As the pressurized hot water effluent in the present invention mainly includes the hemicellulose component, the saccharide solution is purified when C5 saccharide and membrane treatment is performed, in which this pressurized hot water soluble component is subjected to enzymatic saccharification to form pentose (C5 saccharide).

(47) <Enzymatic Saccharification Step>

(48) First, the biomass treated liquid at a high temperature (pressurized hot water effluent) 101A is introduced into the enzymatic saccharification tank 103, to which enzyme 102 is added, and the saccharification is performed by the enzyme reaction in the enzymatic saccharification step.

(49) <Solid-Liquid Separation Step>

(50) Next, the saccharide solution 104 is retained in a first saccharide solution tank 121. After that, the solid residue 111 such as lignin including the water-slightly soluble substances is separated through the solid-liquid separation apparatus 112, and then the saccharide solution 104 is retained in a second saccharide solution tank 122.

(51) <Foreign Substance Removing Step>

(52) Next, remaining foreign substances, such as solid residue 111 such as lignin including the water-slightly soluble substances, are membrane-separated from the saccharide solution 104 through the foreign substance removing unit 113 provided with the MF membrane 113a, and then the saccharide solution 104 is retained in the dilution tank 132.

(53) The saccharide solution 104 including the foreign substances, which have not been removed, is returned to the first saccharide solution tank 121 or the second saccharide solution tank 122.

(54) <Saccharide Solution Dilution Step>

(55) Next, water (RO water) 131 is added to the saccharide solution 104, from which the water-slightly soluble fermentation inhibitory substances have been removed, in the dilution tank 132 to obtain the dilution water 133.

(56) <Saccharide Concentration Step>

(57) Next, water 114 is removed from the dilution water 133 through the water separation unit 116 provided with the RO membrane 115a to obtain the concentrated saccharide solution 115.

(58) From this concentrated saccharide solution 115, various organic feedstocks are obtained in subsequent steps.

(59) In FIG. 1, reference signs M.sub.1 to M.sub.3 show motors which drive stirring means in the enzymatic saccharification tank 103, the first and second saccharide solution tanks 121 and 122, and P.sub.1 to P.sub.3 show liquid sending pumps for sending the saccharide solution 104.

(60) As described above, according to the present Example, the saccharide solution having less impurities can be obtained by removing the water-slightly soluble substances, which are included in the biomass treated liquid at a high temperature 101A generated by decomposition of the biomass feedstock in conditions of a high temperature and a high pressure through the saccharide solution purification means 110; as a result, saccharide solution having a high purity can be obtained and, at the same time, reaction inhibition in subsequent steps, such as fermentation inhibition in a fermentation treatment, can be suppressed.

(61) A first measuring unit of measuring either or both of a turbidity and an absorbance of the concentrated saccharide, solution 115, which has been purified through the saccharide solution purification means 110 and from which the water-slightly soluble substances have been removed, (for example, a turbidimeter and/or a absorption spectrometer) may be included.

(62) In order to monitor a degree of the impurity removal in the concentrated saccharide solution 115 (a degree of clarity of the saccharide solution 104), the turbidity and/or the absorbance are/is measured in the first measurement unit.

(63) As a result, it is possible to confirm the sufficient removal of the water-slightly soluble substances from the concentrated saccharide solution 115, whereby the quality of the saccharide solution 104 can be controlled by the turbidimeter and/or the absorption spectrometer based on the degree of clarity.

Example 2

(64) Next, an alcohol fermentation apparatus that uses biomass as a feedstock according to the present invention will be explained in reference to a view. The same reference signs are applied to the same members as those in the apparatus of Example 1, and the explanations thereof are omitted.

(65) FIG. 2 is a schematic view of a fermentation apparatus that uses biomass as a feedstock according to Example 2.

(66) As illustrated in FIG. 2, an alcohol fermentation apparatus 200 that uses biomass as a feedstock includes the saccharide solution production apparatus 100A using the biomass illustrated in FIG. 1 as the feedstock, and a fermentation tank 140 in which ethanol fermentation is performed using concentrated saccharide solution 115, which has been obtained in the saccharide solution production apparatus 100A using the biomass as the feedstock.

(67) Yeast 142 is added to a fermentation tank 141 of this fermentation tank 140, thereby Obtaining ethanol fermentation liquid 143. The ethanol fermentation liquid 143 is separately subjected to distillation treatment to obtain desired ethanol.

(68) In FIG. 2, reference sign M.sub.5 shows a motor which drives a stirring means in which the stirring is performed in the fermentation tank 141, and P.sub.5 shows a liquid sending pump for sending the ethanol fermentation liquid 143.

(69) According to the present Example, the water-slightly soluble fermentation inhibitory substance can be removed by the saccharide solution purification means 110, and thus the purification is performed in the saccharification treatment, and a good fermentation treatment can be performed in a subsequent alcohol fermentation step.

(70) Here, chemicals based on the saccharide solution may include, for example, lactic acid, and the like, in addition to alcohols. The saccharide solution derived from the biomass, accordingly, can be efficiently utilized as substitutes of chemicals derived from crude oil, which is exhaustible fuel, and feedstock for producing the substitute.

Example 3

(71) Next, an apparatus for producing saccharide solution that uses biomass as feedstock will be explained in reference to a view.

(72) FIG. 3 illustrates a saccharide solution production apparatus according to Example 3, which uses biomass treated liquid at a high temperature, and the explanation is made in reference to the view.

(73) The same reference signs are applied to the same members as those in the apparatus of Example 1, and the explanations thereof are omitted.

(74) As illustrated in FIG. 3, a saccharide solution purification means 110 having a structure different from that in Example 1 illustrated in FIG. 1 is disposed in an apparatus for producing saccharide solution 100B that uses biomass as feedstock.

(75) The saccharide solution purification means 110 illustrated in FIG. 3 includes a saccharide solution purification tank 151 in which saccharide solution 104, which has been taken out from an enzymatic saccharification tank 103, is retained; a solid-liquid separation apparatus 112 which is disposed via a first circulation line L.sub.1 from the saccharide solution purification tank 151, and in which water-slightly soluble substances (solid residue such as lignin) 111 are removed from the taken saccharide solution 104; and a foreign substance removing unit 113 which is disposed via a second circulation line L.sub.2 from the saccharide solution purification tank 151, and which includes a microfiltration (MF) membrane 113a removing water-slightly soluble substances from the taken saccharide solution 104.

(76) In FIG. 3, reference sign M.sub.6 shows a motor which drives a stirring means in the saccharide solution purification tank 151.

(77) According to the saccharide solution purification means 110 of Example 1, the membrane treatment is sequentially performed, but according to the saccharide solution purification means 110 of Example 3, the saccharide solution purification tank 151 has two circulation lines (the first circulation line L.sub.1 and the second circulation line L.sub.2), and thus the treatment appropriate to the concentration condition of the water-slightly soluble substance (solid residue 111 such as lignin) can be performed.

(78) When a content of the water-slightly soluble substance (solid residue 111 such as lignin) is higher than a pre-determined value, accordingly, the treatment using both of the first circulation line L.sub.1 and the second circulation line L.sub.2 is performed to remove the water-slightly soluble substances.

(79) When the content of the water-slightly soluble substance is small, the membrane treatment using the second circulation line L.sub.2 alone can be performed. If necessary, the solid-liquid separation treatment may be performed.

Example 4

(80) Next, a saccharide solution production apparatus that uses biomass as a feedstock according to the present invention will be explained in reference to a view.

(81) FIG. 4 is a schematic view of an apparatus for producing saccharide solution that uses biomass as feedstock according to Example 4. The same reference signs are applied to the same members as those in the apparatuses of Examples 1 and 3, and the explanations thereof are omitted.

(82) As illustrated in FIG. 4, an apparatus for producing saccharide solution 1002 that uses biomass as feedstock includes a saccharide solution purification tank 161 in which the enzymatic saccharification tank 103 in the saccharide solution purification means 110 of Example 4 and the saccharide solution purification tank 151 are integrated.

(83) In the view, reference sign 14, shows a motor which drives a stirring means in the saccharide solution purification tank 161.

(84) According to Example 3, the enzymatic saccharification tank 103 and the saccharide solution purification tank 151 are separately disposed, but according to the present Example, installation space-saving can be contemplated by integrating them.

(85) The saccharide solution production apparatuses 100B and 100C that use biomass of Examples 3 and 4 as a feedstock can also perform various fermentations by disposing, for example, the fermentation tank 140 for performing the ethanol fermentation explained in Example 2.

(86) At that time, good fermentation treatment can be performed in a subsequent fermentation step, because the water-slightly soluble substances are removed.

Experiment Example 1

Identification Test of Water-Slightly Soluble Fermentation Inhibitory Substance

(87) Cooled biomass treated liquid at a high temperature and each of various solvents were mixed in a ratio of 1:9, and the identification was performed using a high-performance liquid chromatography apparatus.

(88) The biomass treated liquid at a high temperature was obtained by hydrothermally decomposing rice straw in a pressurized condition having a temperature of 150 C. using a hydrothermal decomposition apparatus 50 illustrated in FIG. 8, and a pressurized hot water effluent 101A was used.

(89) Acetonitrile, acetone, and methanol, which are all polar solvents, were used as a solvent. Pure water which had been filtered through MILLI-Q (trademark for water filtration systems) was used as Comparative Example.

(90) The procedures are as follows: the cooled biomass treated liquid at a high temperature was mixed with these solvents and water, and the mixtures were allowed to stand in a refrigerator (4 C.) over night.

(91) After that, centrifugal separation (15,000 rpm, 10 minutes, 4 C.) was performed, and supernatant liquid was recovered and the liquid was treated with a filter (0.45 m).

(92) This test liquid (Test 1-1: acetonitrile, Test 1-2: acetone, Test 1-3: methanol, and Test 1-4: pure water) after treated with the filter was analyzed using high-performance liquid chromatography (HPLC).

(93) ODS column (SYNERGI: 4, Hydro-RP80A 4.6250 mm, manufactured by Phenomenex Company) was used as the column.

(94) Water-acetonitrile solvent was used as eluent, which was subjected to a gradient treatment of an acetonitrile concentration of 8% to 100%.

(95) A detection wavelength was set at 220 nm.

(96) The results are shown in FIG. 5 and Table 1.

(97) TABLE-US-00001 TABLE 1 Unknown peak (RT: 22.6 Furfural min, Acetonitrile 55%) (1-1) Acetonitrile 2469009 160550 (1-2) Acetone 2513992 157743 (1-3) Methanol 2428062 145366 (1-4) Pure water 2433687 22275

(98) FIG. 5 illustrates a measured chart of high-performance, liquid chromatography.

(99) As illustrated in FIG. 5 and Table 1, HMF (5-hydroxymethyl furfural: an elution time of around 5.5 minutes) and furfural (an elution time of around 10.5 minutes), which are known fermentation inhibitory substances, were detected in All of Tests 1-1 to 1-4.

(100) On the other hand, an unknown peak component was identified at an elution time of around 22.6 minutes in Tests 1-1 to 1-3. In Test 1-4, the unknown peak component was not detected.

(101) From the above, it was proved that the biomass treated liquid at a high temperature was contained with a water-slightly soluble substance.

Experiment Example 2

Fermentation Test According to Presence or Absence of Water-Slightly Soluble Substance

(102) Fermentation of the saccharification liquid using the cooled biomass treated liquid at a high temperature was performed, and CO.sub.2 decreased amounts in the fermentation were compared.

(103) Pichia stipitis NBRC1687 (standard stock) was used as strain.

(104) A preculture condition was that 10 mL YPD (1% yeast extract, 2% peptone, 2% glucose) was used as a medium, and shaking culture was performed at 30 C. at 120 rpm.

(105) The fermented liquid which had been passed through a filter (0.45 m), and the fermented liquid which had not been passed through the filter (0.45 m) were prepared, and the fermentation was performed.

(106) An ethanol fermentation condition was that 100 mL of fermentation liquid was used and shaking culture was performed at 30 C. at 120 rpm.

(107) The fermentation conditions and the test results are shown in Table 2 and Table 3.

(108) TABLE-US-00002 TABLE 2 Nutrient, Final saccharide concentration Yeast Test Filter ex.* CSL* Glucose Xylose No Solvent (0.45 nm) (%) (%) (%) (%) (2-1) Saccharide Absent 0 0 7.2 8.5 (2-2) solution Present (2-3) from Absent 1.0 1.0 (2-4) biomass Present treated at high temperature (C5 saccharide) Condition: 100 mL of fermented liquid, 30 C., 120 rpm Shaking culture *Yeast ex.: Yeast extract *CSL: Corn steep liquor Strain used: Pichia stipitis NBRC1687 (standard strain) Pre-culture conditions: Medium 10 mL YPD (1% Yeast ex.*, 2% Peptone, 2% Glucose) Conditions 30 C., 120 rpm Shaking culture

(109) TABLE-US-00003 TABLE 3 Final concentration of Glucose, Xylose, or EtOH Concentration Glucose Xylose EtOH Fermentation Test No. (%) (%) (%) efficiency (%) (2-1) 5.9 7.2 N.D. 0 (2-2) 3.7 7.1 0.9 11 (2-3) 5.4 7.4 N.D. 0 (2-4) N.D. 5.0 3.0 38 N.D.: Not Detected

(110) FIG. 6 and FIG. 7 are charts illustrating the results of decreased amounts of carbon dioxide in the ethanol fermentation test, in which the horizontal axis shows time, and the vertical axis shows a CO.sub.2 decreased amount.

(111) As illustrated in FIG. 6 and FIG. 7, the fermentation test showed that the difference in the decreased amount of carbon dioxide appeared after about 90 hours passed in Test 2-2 (no nutrient, presence of the filter) compared to Test 2-1 (no nutrient, no filter).

(112) The difference in the decreased amount of carbon dioxide appeared after about 20 hours passed and after that the decreased amount was remarkably increased in Test 2-4 (presence of nutrient, presence of the filter) compared to Test 2-3 (presence of nutrient, no filter).

(113) The fermentation efficiency was 11% in Test 2-2, and the fermentation efficiency was 38% in Test 2-4.

(114) It was confirmed, therefore, that when the treatment with a filter (0.45 m) was performed, the water-slightly soluble fermentation inhibitory substances are removed, resulting in performance of good fermentation.

(115) As described above, it was proved that when the water-slightly soluble substances were removed by the saccharide solution purification means, the saccharide solution including less impurities could be obtained, and the fermentation inhibition could be remarkably suppressed in the alcohol fermentation treatment in the subsequent steps.

REFERENCE SIGNS LIST

(116) 50 hydrothermal decomposition apparatus 51 biomass feedstock 52 biomass supplying unit 53 apparatus body 54 conveying screw means 55 pressurized hot water 56 biomass solid component 57 biomass discharging apparatus 90 cooling means 100A to 100C apparatus for producing saccharide solution that uses biomass as feedstock 101A biomass treated liquid at a high temperature (pressurized hot water effluent) 101B cooled treated liquid 102 enzyme 103 enzymatic saccharification tank 104 saccharide solution 110 saccharide solution purification means 140 fermentation tank 200 alcohol fermentation apparatus that uses biomass as a feedstock