A PRETREATMENT PROCESS OF LIGNOCELLULOSIC BIOMASS

Abstract

The present invention relates to a pretreatment process of lignocellulosic biomass, comprising contacting lignocellulosic biomass with pretreatment solution in a temperature ranging from an ambient temperature to 100° C. and a pressure of at least 20 bars, wherein the pretreatment solution comprising water, water-miscible organic solvent and alkali having a concentration of 5 to 15% w/v if the alkali is solid or 5 to 15% v/v if the alkali is liquid. Said pretreatment process operates in mild condition or at low temperature to be specified with the removing of lignin but not destroying cellulose and increasing desired sugar yield.

Claims

1. A pretreatment process of lignocellulosic biomass comprising contacting lignocellulosic biomass with pretreatment solution in a temperature ranging from an ambient temperature to 100° C. and a pressure of at least 20 bars, wherein the pretreatment solution comprising water, water-miscible organic solvent and alkali having a concentration of 5 to 15% w/v if the alkali is solid or 5 to 15% v/v if the alkali is liquid.

2. The process according to claim 1, wherein the alkali having a concentration of 5-12% w/v if the alkali is solid or 5-12% v/v if the alkali is liquid.

3. The process according to claim 1, wherein the temperature is in the range of 50 to 80° C.

4. The process according to claim 1 or 2, wherein the water-miscible organic solvent is selected from acetone, ethanol, ethyl acetate, or a mixture thereof.

5. The process according to claim 4, wherein the water-miscible organic solvent is acetone.

6. The process according to claim 1, wherein a concentration of the water-miscible organic solvent is in a range of 70 to 95% v/v.

7. The process according to claim 6, wherein the concentration of the water-miscible organic solvent is in the range of 80 to 90% v/v.

8. The process according to claim 1 or 2, wherein the alkali is selected from sodium hydroxide, ammonium hydroxide, triethylamine, or a mixture thereof.

9. The process according to claim 8, wherein the alkali is sodium hydroxide.

10. The process according to claim 1, wherein pretreatment time of lignocellulosic biomass is not more than 60 minutes.

11. The process according to claim 10, wherein the pretreatment time of lignocellulosic.

12. The process according to claim 1, wherein said process further comprising stirring at a speed rate ranging from 250 to 300 rpm.

13. The process according to claim 1, wherein lignocellulosic biomass in dry form is in a range of 2 to 9% w/v of the pretreatment solution.

14. The process according to claim 1, wherein lignocellulosic biomass is selected from rice straw, bagasse, corncob, com fiber, pineapple husk, or bamboo.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0010] FIG. 1 shows an effect of ammonium hydroxide in various solvents (distilled water (DI); acetone; ethanol (EtOH); and ethyl acetate (EA): to sugar yield from digestion of enzymatic pretreatment solid.

[0011] FIG. 2 shows an effect of triethylamine in various solvents (DI; acetone; EtOH; and EA) to sugar yield from digestion of enzymatic pretreatment solid.

[0012] FIG. 3 shows an effect of sodium hydroxide in variuos solvents (DI; acetone; EtOH; and EA) to sugar content from digestion of enzymatic pretreatment solid.

[0013] FIG. 4 shows an effect of pretreatment process temperature to sugar yield from digestion of enzymatic pretreatment solid.

[0014] FIG. 5 shows an effect of organic solvent content in pretreatment process to sugar yield from digestion of enzymatic pretreatment solid.

[0015] FIG. 6 shows an effect of sodium hydroxide in pretreatment process to sugar yield from digestion of enzymatic pretreatment solid.

[0016] FIG. 7 shows an effect of pretreatment process time to sugar yield from digestion of enzymatic pretreatment solid, wherein alkali with a concentration of 5% w/v was used in pretreatment process.

[0017] FIG. 8 shows an effect of pretreatment process time to sugar yield from digestion of enzymatic pretreatment solid, wherein alkali with a concentration of 7% w/v was used in pretreatment process.

[0018] FIG. 9 shows an effect of starting solid content in system to sugar yield from digestion of enzymatic pretreatment solid.

DETAILED DESCRIPTION OF THE INVENTION

Definition

[0019] Technical terms or scientific terms used herein, unless stated otherwise, have their definitions as known in persons skilled in the art.

[0020] Any tool, equipment, method, or chemical mentioned herein shall mean tool, equipment, method, or chemical being used or practiced generally by a person skilled in the art of this field unless stated otherwise that they are tools, equipment, methods, or chemicals specific only in this invention.

[0021] Use of singular nouns or pronouns when used with “comprising” in claims and/or specification means “one” and will also include “one or more”, “at least one”, and “one or more than one”.

[0022] Throughout this application, the term “about” used to indicate any value that is appeared or expressed herein may be varied or deviated, which the variation or deviation may occur from the error of instruments and methods used to determine various values.

[0023] An objective of this invention is to develop a pretreatment process of lignocellulosic biomass with pretreatment solution comprising water, water-miscible organic solvent and alkali having a concentration of 5 to 15% w/v if the alkali is solid, or 5 to 15% v/v if the alkali is liquid. The pretreatment process operates in mild condition or at low temperature.

[0024] Hereafter, invention embodiments are shown without any purpose to limit any scope of the invention.

[0025] This invention relates to the pretreatment process of lignocellulosic biomass comprising contacting lignocellulosic biomass with pretreatment solution in a temperature ranging from an ambient temperature to 100° C. and a pressure of at least 20 bars, wherein the pretreatment solution comprising water, water-miscible organic solvent and alkali having a concentration of 5 to 15% w/v if the alkali is solid, or 5 to 15% v/v if the alkali is liquid. Preferable, the alkali has a concentration of 5 to 12% w/v if the alkali is solid or 5 to 12% v/v if the alkali is liquid.

[0026] In one aspect, the water-miscible organic solvent is selected from acetone, ethanol, ethyl acetate, or a mixture thereof. Most preferable, said organic solvent is acetone.

[0027] Preferably, a concentration of the water-miscible organic solvent is in a range of 70 to 95% v/v. More preferable is in a range of 80 to 90% v/v.

[0028] Preferred alkali is selected from sodium hydroxide, ammonium hydroxide, triethylamine, or a mixture thereof. Most preferable, said alkali is sodium hydroxide.

[0029] In another aspect of the invention, a temperature of the pretreatment process of lignocellulosic biomass is ranging from an ambient temperature to 100° C. Preferable the temperature is in the range of 50 to 80° C.

[0030] In another aspect of the invention, a pretreatment time of lignocellulosic biomass is not more than 60 minutes. Preferable it is 5 to 30 minutes starting after system reaches targeted temperature.

[0031] The pretreatment process of lignocellulosic biomass may further comprise stirring at a speed rate ranging from 250 to 300 rpm.

[0032] The lignocellulosic biomass in dry form is in a range of 2 to 9% w/v of the pretreatment solution. The lignocellulosic biomass may be selected from rice straw, bagasse, corncob, corn fiber, pineapple husk, or bamboo.

[0033] The followings are examples of a preparation of lignocellulosic biomass before pretreatment process, a pretreatment process according to the invention, and an analysis of sugar yielded from pretreatment.

[0034] The preparation step of biomass could be done by the reduction of rice straw size using cutting machine and sizing using 2 mm mesh.

[0035] The pretreatment of lignocellulosic biomass obtained from biomass preparation step was performed in 600 mL, high pressure stainless steel reactor (Parr reactor 4848, Parr instrument, USA) equipped with thermometer inside. The pretreatment process comprises of feeding of lignocellulosic biomass which was 4 grams of rice straw, 160 mL acetone, and 200 mL of 5% w/v of sodium hydroxide solution. Then, nitrogen gas was purged into the system to have initial pressure of 20 bars. Then, the system was heated at about 80° C. for 30 minutes starting after the system had reached targeted temperature. After that, the pretreatment process was stopped by reducing temperature such as submerging reactor into water or any other coolants. Then, filtration had been performed to separate solid and liquid parts. The solid part was put into the oven at 60° C.

[0036] The analysis of sugar content yielded from the pretreatment process was done. The above obtained solid part was digested by adding 20 FPU/g of cellulose enzyme (Accellerase 1500, Genecor, USA) in 50 mmol sodium acetate buffer with pH of 4.8. The said solid part was then incubated at a temperature of 50° C. for 72 hours with tube shaking at a speed rate of about 30 rpm. The liquid part was collected as sample for quantitative of reducing sugar using dinitrosalicylic acid method (Miller 1959).

[0037] Note: FPU of cellulase enzymes was obtained from the digestion of filter paper obtained from the analysis according to standard method (Adney and Baker 1996).

[0038] The quantitative of reducing sugar after enzymatic digesting could be determined using high performance liquid chromatography (HPLC) method (Water e2695, Water, USA), equipped with differential refractometer, and Aminex HPX-87H column (Bio-Rad, USA). The compositions in liquid part were treated with acid and digested by a method according to NREL standard. Glucose yield was calculated into theoretical percentage comparing to cellulose content in starting biomass in dry form, which was 358 mg/g (x1.11).

[0039] Effect of the Pretreatment Using Pretreatment Solutions Having Different Alkali

[0040] 4 grams of rice straw, equally to 2% w/v was exposed to a pretreatment solution comprising 160 mL acetone and sodium hydroxide, ammonium hydroxide, and triethylamine wherein the concentration of the alkali was 5% w/v (for sodium hydroxide), or 5% v/v (for ammonium hydroxide and triethylamine). The pretreatment process was performed at a temperature around 80° C. for about 30 minutes at initial pressure around 20 bars with a speed rate of stirring at 250 rpm.

[0041] From table 1 and FIGS. 1, 2, and 3, it was found that the use of different alkali catalysts increases solid digestibility of pretreatment process comparing to without alkali catalyst. The process without catalyst provided maximum reducing sugar from enzymatic digestion of 146 mg/g comprising 131 mg of glucose per gram of pretreated dry biomass which is equal to 26.7% of glucose theoretical yield. The use of sodium hydroxide provided the best pretreatment effect, giving 75.6% of glucose theoretical yield, provided maximum reducing sugar from enzymatic digestion of 865 mg/g comprising 834 mg of glucose per gram of pretreated dry biomass, whereas the use of ammonium hydroxide and triethylamine provided 34.3% and 33.0% of glucose theoretical yields respectively. This test showed the effect of alkali catalysts in the pretreatment process of lignocellulosic biomass using a mixture of organic solvent. These results were compared to the use of hydrogen peroxide as an oxidizing agent in the pretreatment. Results showed that hydrogen peroxide could react with acetone to become acetone peroxide which was very reactive and could lead to explosion which was not suitable to be used with acetone.

TABLE-US-00001 TABLE 1 Effect of pretreatment solutions, pretreatment of ligno- cellulosic biomass and different alkali catalysts in acetone to sugar content from enzymatic digestion Reducing sugar Glucose (mg/g (mg/g % weight Theoretical Type of alkali in pretreated pretreated loss of percentage pretreatment solution biomass) biomass) biomass of glucose — 146 131 19 26.7 Sodium hydroxide 856 834 64 75.6 Ammonium hydroxide 215 192 29 34.3 Triethylamine 217 166 21 33.0

[0042] Effect of Temperature to the Pretreatment Process of Lignocellulosic Biomass

[0043] This experiment studied the effect of sodium hydroxide in the pretreatment process of lignocellulosic biomass using acetone in various target temperatures ranging from about 50 to 100° C. 2% w/v of rice straw in dry form of 2% w/v was expose to pretreatment solution comprising 160 mL acetone and 5% weight by final volume of sodium hydroxide, pretreatment process was performed for 30 minutes at initial pressure of 20 bars, with stirring speed at 250 rpm.

[0044] From table 2 and FIG. 4, it was found that pretreatment at about 50° C. gave highest glucose of 85.7% theoretically yield comparing to cellulose in native biomass. Reducing sugar obtained from enzymatic digestion was 815 mg, containing 774 mg of glucose per g of pretreated dry biomass. Sugar content in the term of glucose theoretically yield percentage obtained from the use of sodium hydroxide as catalyst was higher than without catalyst and the use of ammonium hydroxide and triethylamine under same conditions at 80 to 100° C.

TABLE-US-00002 TABLE 2 Effect of temperature to sugar content obtained from solid enzymatic digestion after pretreatment Reducing Glucose Weight loss Theoretical Temperature sugar (mg/g (mg/g percentage of percentage of (° C.) pretreated) pretreated) biomass glucose yield 50 815 774 56 85.7 60 870 832 61 81.6 70 872 839 62 80.2 80 856 834 64 75.6 90 864 833 69 64.9 100 865 834 70 63.0

[0045] Effect of the Content of Water-Miscible Organic Solvent

[0046] The effect of the amount of water-miscible organic solvent in pretreatment process using sodium hydroxide as a catalyst was studies. 2% w/v of rice straw in dry form was exposed to pretreatment solution comprising acetone from 160 to 180 mL and sodium hydroxide. Then water was added until the volume of the solution is 200 mL, so that acetone concentration was 80 to 90% v/v. The concentration of alkali was 5% w/v. The pretreatment process was performed at temperature around 80° C. for about 30 minutes at initial pressure of 20 bars with stirring speed at about 250 rpm.

[0047] From table 3 and FIG. 5, it was found that the concentration of acetone effected sugar yield from enzymatic digestion step. Acetone having a concentration of 85% v/v gave highest glucose theoretically yield of 80.7% comparing to cellulose content in native biomass. Reducing sugar from enzymatic digestion step was 902 mg with 867 mg of glucose per g of pretreated dry biomass.

TABLE-US-00003 TABLE 3 Effect of acetone ratio to sugar content obtained from enzymatic digestion Reducing Weight Theoretical sugar Glucose loss yield Acetone (mg/g (mg/g percentage percentage concentration pretreated pretreated of of percentage biomass) biomass) biomass glucose 80 856 834 64 75.6 85 902 867 63 80.7 90 933 899 68 72.4

Effect of Alkali Concentration

[0048] Effect of concentration of sodium hydroxide in the pretreatment process of rice straw using acetone was studied in this experiment. 2% w/v of dry rice straw was exposed to the pretreatment solution comprising 160 mL acetone and sodium hydroxide at various concentrations ranging from 5 to 10% w/v. The pretreatment process was performed at temperature around 80° C. for about 30 minutes at initial pressure of 20 bars with stirring speed at about 250 rpm.

[0049] From table 4 and FIG. 6, it was found that increasing of sodium hydroxide concentration gave higher sugar yield from enzymatic digestion step. Sodium hydroxide at concentration of 10% w/v gave highest glucose at 83.6% theoretically yield comparing to cellulose content in native biomass. Reducing sugar from enzymatic digestion step was 982 mg with 949 mg of glucose per g of pretreated dry biomass.

TABLE-US-00004 TABLE 4 Effect of sodium hydroxide concentration to sugar obtained from enzymatic digestion Reducing Weight Theoretical Sodium sugar Glucose loss yield hydroxide (mg/g (mg/g percentage percentage concentration pretreated pretreated of of (%) biomass) biomass) biomass glucose 5 856 834 64 75.6 7.5 916 877 66 75.0 10 982 949 65 83.6

[0050] Effect of Time to the Pretreatment Process of Lignocellulosic Biomass

[0051] This experiment studied the effect of sodium hydroxide in pretreatment process of lignocellulosic biomass using acetone at pretreatment of 0 to 60 minutes. 2% w/v of dry rice straw was exposed to the pretreatment solution comprising 180 mL acetone and sodium hydroxide at concentration from 5 to 7% w/v. The pretreatment process was performed at temperature around 80° C. at initial pressure of 20 bars with stirring speed at about 250 rpm. Time was counting after the system reached its targeted temperature.

[0052] From table 5 and 6 and FIGS. 7 and 8, it was found that increasing of time gave higher sugar yield from enzymatic digestion step. However, when comparing with cellulose content in native biomass, it was found that at alkali concentration of 5%, pretreatment at 5 minutes gave highest glucose of 91.9% theoretically yield comparing to cellulose content in native biomass. Reducing sugar from enzymatic digestion step was 945 mg with 913 mg of glucose per g of pretreated dry biomass. Whereas when alkali concentration was 7%, pretreatment at 30 minutes gave highest glucose of 96.3% theoretically yield comparing to cellulose content in native biomass. Reducing sugar from enzymatic digestion step was 962 mg with 933 mg of glucose per g of pretreated dry biomass.

TABLE-US-00005 TABLE 5 Effect of time to sugar obtained from enzymatic digestion (at alkali concentration of 5% w/v) Reducing Weight Theoretical sugar Glucose loss yield (mg/g (mg/g percentage percentage Time pretreated pretreated of of (minute) biomass) biomass) biomass glucose 0 873 832 57 90.0 5 945 913 60 91.9 10 949 913 63 85.0 20 943 909 64 82.3 30 948 911 67 75.7 60 953 919 64 83.3

TABLE-US-00006 TABLE 6 Effect of time to sugar obtained from enzymatic digestion (at alkali concentration of 7% w/v) Reducing Weight Theoretical sugar Glucose loss yield (mg/g (mg/g percentage percentage Time pretreated pretreated of of (minute) biomass) biomass) biomass glucose 0 779 730 51 90.0 5 902 874 58 92.4 10 925 894 59 92.2 20 950 925 63 86.1 30 962 933 59 96.3 60 974 950 61 93.2

[0053] Effect of Biomass Content

[0054] In this experiment, effect of biomass content in starting solid matter in the pretreatment process using pretreatment solution comprising sodium hydroxide and acetone was studied. 2 to 9% w/v of dry rice straw was exposed to the pretreatment solution comprising 180 mL acetone and sodium hydroxide at concentration of 5% w/v. The pretreatment process was performed at temperature around 80° C. for about 30 minutes at initial pressure of 20 bars with stirring speed at about 250 rpm.

[0055] From table 7 and FIG. 9, it was found that increasing of starting solid content affected the sugar yield from enzymatic digestion step. The use of 6% w/v of starting solid matter gave highest glucose of 86.2% theoretically yield comparing to cellulose content in native biomass. Reducing sugar from enzymatic digestion step was 878 mg with 835 mg of glucose per g of pretreated dry biomass.

TABLE-US-00007 TABLE 7 Effect of starting solid matter in the system to sugar yield obtained from enzymatic digestion Reducing Weight Theoretical sugar Glucose loss yield (mg/g (mg/g percentage percentage Starting solid pretreated pretreated of of matter (%) biomass) biomass) biomass glucose 2 938 903 68 72.7 4 948 911 67 75.7 6 878 835 59 86.2 8 822 762 59 78.6 9 865 811 60 81.6

Best Mode of the Invention

[0056] Best mode or preferred embodiment of the invention is as provided in the description of the invention.