結合超音波與離子液體於木質纖維素生質原料之預處理及其酵素水解反應動力學之研究

楊珺堯

doi:10.6845/NCHU.2015.01255

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結合超音波與離子液體於木質纖維素生質原料之預處理及其酵素水解反應動力學之研究

Combination of Ultrasound with Ionic Liquid Pretreatment and Kinetics of Enzymatic Hydrolysis for Lignocellulosic Biomass

楊珺堯 , Ph.D　　Advisor：方繼　　

英文 DOI： 10.6845/NCHU.2015.01255

稻稈；稻殼；預處理；離子液體；超音波；酵素水解；動力學； rice straw ； rice hull ； pretreatment ； ionic liquid ； ultrasound ； enzymatic hydrolysis ； kinetics

Abstract │ Reference (122) │ Altmetrics

Abstract 〈TOP〉

稻稈和稻殼是台灣常見且豐富的農業廢棄物，其富含木質纖維素，並具有作為生產生物燃料的原料之潛力。針對有效使用木質纖維素生質原料而言，稻稈與稻殼必須經由一個有效的預處理方法來進行預處理，以減少對後續的酵素水解反應之阻礙。本研究開發一個結合超音波與離子液體的有效預處理方法，並將此方法應用於稻稈和稻殼的預處理製程上，對於在不同的預處理條件下對稻稈進行降解以及酵素水解之製程探討。此外，發展出一套簡化的動力學模式來描述稻稈與稻殼的酵素水解反應在非均勻相系統中之動力學，並將此模式稱為" Michaelis 阻礙模式" (impeded Michaelis model)。
本研究進行預處理前後之稻稈與稻殼的形態與特徵結構之探討，藉由場發射電子掃描顯微鏡 (FE-SEM)、X光能量散譜儀 (EDS)、原子力顯影功能 (AFM)、霍式紅外光譜共焦平面陣列顯影功能 (FPA-FTIR)、高解析X光繞射儀 (HR-XRD)、元素分析儀 (EA)來觀察預處理前後之變化。本研究對生質原料的不同的預處理條件進行探討，且以不同的基質藉由纖維分解酵素(cellulase from Trichoderma reesei ATCC 26921)來進行酵素水解反應，並搭配超音波(300 W/40 kHz)輔助酵素水解作用。
本研究使用之不同的離子液體包括1-ethyl-3-methylimidazolium ethylsulfate ([EMIM][EtSO4])、trihexyl(tetradecyl)phosphonium decanoate (THTDPD) 與生物-鹼性離子液體 choline hydroxide ([Ch][OH])，將其應用於稻稈的預處理，使其表面結構在預處理後被破壞並改變其組成。在超音波輔助酵素水解的探討上，以CHRS (稻稈以超音波與[Ch][OH]預處理之基質) 為水解基質並在超音波的輔助下進行240分鐘的酵素水解作用，其總還原醣產率(TRS)之轉換可高達96.22%。稻稈以[Ch][OH]進行預處理表現出較佳的效率，當搭配超音波可以有效地應用在稻稈之預處理與酵素水解作用。
以超音波與[Ch][OH]對稻稈進行預處理後，發現其結晶度增加及官能基團之變化，因而顯著地提高酵素水解的效率。本研究以" Michaelis 阻礙模式"來描述稻稈的酵素水解反應，並根據實驗觀察，提出了隨時間變化之衰減係數(Time-dependent decay coefficient)作為阻礙反應的因子，可獲得初始速率常數與無效係數來說明酵素和基質間之交互作用，並證實此模式的可應用性。
針對稻殼以[Ch][OH]與超音波預處理之酵素水解進行探討，經預處理後的稻殼，可移除二氧化矽，木質素與灰分，使其網絡結構呈現嚴重腐蝕，並增加有機部分的暴露，得以提高酵素水解反應。在超音波輔助酵素水解的情況下，在只有3小時的水解時間，稻殼預處理後之TRS產率為43.5％，並顯著高於未經處理的稻殼（24.0％），且" Michaelis 阻礙模式"也一樣可成功地用於描述稻殼的酵素水解反應。

Parallel Abstract 〈TOP〉

Rice straw and rice hull, known as the common and abundant agricultural wastes in Taiwan, contain an abundant lignocellulosic biomass and have the potential to serve as raw materials for the production of biofuel. For the effective use of lignocellulosic biomass, rice straw and rice hull have to be pretreated by an efficient pretreatment process to eliminate the obstacles for the subsequent hydrolysis by enzymes. In this dissertation, the effective pretreatment method for rice straw and rice hull by using ultrasound and ionic liquids was developed, and the degradation of rice straw and rice hull under different processes of pretreatment and enzymatic hydrolysis was investigated. A simplified kinetic model, called the "impeded Michaelis model" was developed to describe the kinetics of enzymatic hydrolysis in a heterogeneous system.
The morphological and characteristic structures of rice straw and rice hull before and after the pretreatment were analyzed by field emission scanning electron microscope (FE-SEM), X-ray energy dispersive spectrometer (EDS), focal plane array-Fourier transform infrared spectroscopy (FPA-FTIR), high resolution X-ray diffractometer (HR-XRD), and elemental analyzer (EA). Different pretreatment conditions for the biomass were explored. Various substrates were used to conduct enzymatic hydrolysis by cellulase from Trichoderma reesei ATCC 26921 with and without ultrasonic irradiation (300 W/40 kHz).
Various ionic liquids, including 1-ethyl-3-methylimidazolium ethylsulfate ([EMIM][EtSO4]), trihexyl (tetradecyl) phosphonium decanoate (THTDPD), and a bio-based basic ionic liquid choline hydroxide ([Ch][OH]) were used to pretreat rice straw under ultrasound, and resulting in the surface structure eroded and composition changes after the pretreatment. In ultrasound-mediated enzymatic hydrolysis for rice straw using cellulase, the yield of total reducing sugar (TRS) converted from the rice straw after pretreatment with [Ch][OH] and ultrasound (denoted as CHRS) was up to 96.22% at 240 min and was greater than that from the other substrates; the TRS yield for CHRS with ultrasound was 19.5% greater than that without irradiation. Rice straw pretreated with [Ch][OH] showed the highest efficiency among the tested ILs, and ultrasound can be applied effectively in rice straw pretreatment and enzymatic hydrolysis.
The increase of crystallinity and functional groups changes were observed after the pretreatment on rice straw with [Ch][OH] under ultrasound, thus significantly enhancing the enzymatic hydrolysis by cellulase. A simplified kinetic model, the "impeded Michaelis model" with the impeded reaction of enzyme was developed to successfully describe the enzymatic hydrolysis of rice straw in a heterogeneous system. Based on the experimental observation, a time-dependent decay coefficient for the impeded reaction was proposed. The initial observed rate constants as well as ineffective coefficients were obtained to elucidate the interaction of enzyme and substrate, and the applicability of the simplified model was demonstrated.
For the enzymatic hydrolysis of rice hull by the ultrasonic pretreatment using [Ch][OH] was investigated. The network structure of rice hull after the pretreatment was seriously corroded and punched to expose the accessible organic region by removing silica, lignin and ash to enhance the hydrolysis reaction. In ultrasound-assisted enzymatic hydrolysis by cellulase, TRS yield from rice hull after the pretreatment was 43.5% at only 3 h of hydrolysis time, and was greater than that from untreated rice hulls (24.0%). The "impeded Michaelis model" was also successfully used to describe the enzymatic hydrolysis of rice hull as well.

Reference ( 122 ) 〈TOP〉

Agbor, V.B., Cicek N., Sparling R., Berlin A., Levin D.B., 2011. Biomass pretreatment: Fundamentals toward application. Biotechnol. Adv. 29, 675 - 685.
連結：
Ahn, Y., Lee, S.H., Kim, H.J., Yang, Y.H., Hong, J.H., Kim, Y.H., Kim, H., 2012. Electrospinning of lignocellulosic biomass using ionic liquid. Carbohydr. Polym. 88, 395- 398.
連結：
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連結：
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連結：
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連結：

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