生物質(zhì)制燃料乙醇

石油化工2007年第36卷第2期PETROCHEMICAL TECHNOLOGY107·特約述評生物質(zhì)制燃料乙醇陳輝,陸善祥(華東理工大學(xué)聯(lián)合化學(xué)反應工程研究所,上海200237要】生物質(zhì)制燃料乙醇具有重要環(huán)保意義是一科極具前景的石油可替代資源生產(chǎn)工藝。目前,以植物纖維素為原料生產(chǎn)燃料乙醇的成本仍較髙。綜述了近幾年來(lái)生物質(zhì)制燃料乙醇在預處理工藝、水解液發(fā)酵抑制物的脫除方法、水解和發(fā)酵工藝、纖維素酶和乙醇發(fā)酵基因工程菌領(lǐng)域的研究進(jìn)展,介紹了國外相關(guān)的大的生物質(zhì)制燃料乙醇項目,概述了生物量全利用的意義,展望了生物質(zhì)制燃料乙醇的未來(lái)發(fā)展方向[關(guān)鍵詞]燃料乙醇;生物質(zhì);纖雄素;基囚工程;發(fā)酵[文章編號]1000-8144(2007)02-0107-11[中圖分類(lèi)號]TQ5172文獻標識碼]AProgress in Production of Fuel-Ethanol from Biomass ResourcesChen Hui, Lu shanxiangUNILAB of East China University of Science and Technology, Shanghai 200237, China)[ Abstract] Production of fuel-ethanol from biomass resources is environment -friendly and is apromising substitutive technology for limited crude oil resource. Current cost for production of ethanolfrom cellulose plant is still high. The progresses in technology, including pretreatment of materialsremoval of fermentation inhibitors in hydrolysate, hydrolysis and fermentation technology, andenetically engineered strains for cellulase production and ethanol fermentation were reviewed. Theorlds important projects for utilization of biomass were briefly described and future development forproduction of fuel-ethanol from biomass was discussedI Keywords fuel ethanol; biomass; cellulose genetic engineering; fermentation燃料乙醇是指作為燃料添加到汽油、柴油中的為原料的燃料乙醇生產(chǎn)成本約4000元/t。燃料乙乙醇,可部分替代石油,緩解石油資源短缺。從生物醇的生產(chǎn)成本太高,不足以與汽油競爭,目前各國普質(zhì)光合作用、生物質(zhì)制燃料乙醇到乙醇燃燒的過(guò)程遍采用補貼或稅收優(yōu)惠的方式攴持燃料乙醇的生在自然界形成了℃O2的閉合循環(huán),可緩解地球的溫產(chǎn)。植物纖維素資源是地球上最豐富和廉價(jià)的可再室效應,燃料乙醇還可降低汽車(chē)尾氣污染,因而燃料生資源,通過(guò)生物化學(xué)法和熱化學(xué)法(熱解、氣化乙醇具有重要的生態(tài)環(huán)保意義。汽油中乙醇的體液化和超臨界抽提)可生產(chǎn)燃料乙醇,但生產(chǎn)成本積分數低于10%-15%時(shí),不僅不需對現有汽車(chē)發(fā)很高。近年來(lái)隨著(zhù)化工過(guò)程、生物技術(shù)基因工程的動(dòng)機進(jìn)行改進(jìn),且汽油還具有辛烷值高和抗爆性好發(fā)展,燃料乙醇的生產(chǎn)成本逐漸降低。從長(cháng)遠來(lái)看,的優(yōu)點(diǎn)因而燃料乙醇已在世界許多國家廣泛應用,生物化學(xué)法具有良好的發(fā)展和應用前景。推廣力度不斷加大。本文綜述了近幾年來(lái)生物質(zhì)制燃料乙醇在工燃料乙醇生產(chǎn)的主要問(wèn)題是原料成本高。以含藝、糖作物或谷物為原料時(shí),原料成本占燃料乙醇生產(chǎn)相關(guān)成本的60%左右。巴西以甘蔗為原料的燃料乙醇*H中國煤化工究進(jìn)展,介紹了國外CNMHG目,腰望了生物質(zhì)生產(chǎn)成本約02美元,美國以玉米為原料的燃料收稿日期2009-2修改稿日期]1206-10-16乙醇生產(chǎn)成本約0.3美元/L,歐洲以小麥為原料的作者簡(jiǎn)介」陳輝(1974-),女,山東省濰坊市人碩士,講師。聯(lián)系燃料乙醇生產(chǎn)成本約0.48美元/L,我國以玉米陸善樣,電話(huà)021-642100,電郎hshx@sohu.com石油化PETROCHEMICAL TECHNOLOGY2007年第36卷制燃料乙醇的發(fā)展方向結合,不僅部分溶解纖維素,而且可破壞纖維素的物1原料的種類(lèi)不斷豐富理結構。 Foody等采用一系列帶V形齒槽的輥子輾壓、剪切潤濕的木質(zhì)纖維素原料(適用于打漿可提供糖的資源都可用作乙醇發(fā)酵的原料。除泵送),漿液先脫除一部分水和水溶性物質(zhì)后,再在了含糖和含淀粉作物,植物纖維素原料中的纖維素160~280℃下對漿液進(jìn)行稀酸水解預處理?？伤鉃槠咸烟?半纖維素可水解為戊糖和己糖。2.2.2堿預處理法由于耕地有限,可開(kāi)發(fā)劣質(zhì)土地種植高產(chǎn)、高糖、耐堿性試劑可采用 NaOH. Ca(OH)2、氨等。堿金堿、耐旱的經(jīng)濟作物。此外,植物纖維素是自然界產(chǎn)屬氫氧化物水溶液(質(zhì)量分數為20%-40%)9的量最大的原料,主要有農產(chǎn)品廢棄物和草木。我國堿性強,通過(guò)溶解脫除木質(zhì)素提高原料的酶可及度。是農業(yè)大國,據測算,每年農田秸桿資源的一半轉化NaOH還具有潤脹纖維素的作用,可降低纖維素的為燃料乙醇燃料乙醇的消費量將超過(guò)年汽油消費結晶度,易于纖維素酶的水解20。量的1.2倍以上2?；诮档统杀竞吞幚韽U物的考Ca(OH)2的堿性較強堿弱,對于玉米秸桿,較慮,纖維素原料的種類(lèi)還擴展到含有纖維素的城市優(yōu)化的處理條件為:在氧氣的存在下,55℃處理28廢棄物或工業(yè)低價(jià)值副產(chǎn)品,如建筑垃圾”、城市d,可脫除幾乎所有的乙?；?、87.5%的木質(zhì)素,約固體垃圾6、酒精廠(chǎng)釜餾物、玉米谷粒濕磨91.3%的纖維素和51.8%的半纖維素水解為葡萄副產(chǎn)物纖維、軋棉廢棄物、橄欖油提煉殘糖和木糖,每克原料僅消耗0.073gCa(OH)2,且預[14處理液不含有乙醇發(fā)酵抑制物21。2纖維素原料的預處理氨不僅價(jià)格便宜,還可回收循環(huán)利用,因而被廣泛應用。氨回收滲濾(ARP)法具有潤脹和脫木質(zhì)纖維素分子內和分子間存在氫鍵,聚集態(tài)結構素的作用,處理玉米秸桿時(shí),可溶解約50%的半纖復雜且結晶度高反應活性低;天然纖維素原料中含維素,保留約92%以上的纖維素,且纖維素基本的有的木質(zhì)素和半纖維素在空間上可阻礙甚至封閉纖晶體結構沒(méi)有明顯變化。熱水處理和ARP兩步預維素分子與酶或化學(xué)試劑的接觸,酶可及度差,更增處理:玉米秸桿可在熱水處理過(guò)程中脫除84%的半維素消除空間障礙;降低纖維素的聚合度和結晶素預處理產(chǎn)物中含有79%的纖維素《%的木質(zhì)加了水解的難度。通過(guò)預處理可脫除木質(zhì)素和半纖纖維素,并在后續的ARP過(guò)程中脫除75%的木質(zhì)度,從而有利于纖維素的有效利用。預處理可采用2.2.3濕法氧化預處理法物理法、化學(xué)法、水熱化學(xué)法和生物法,或幾種方法濕法氧化是指水、氧氣、弱堿或弱酸在高溫相結合,此外還有電解法5、超聲法等。定壓力下氧化降解生物質(zhì)的過(guò)程。 Lessens等對2.1物理法高木質(zhì)素含量的木材廠(chǎng)廢料進(jìn)行濕法氧化預處理物理法:可破壞纖維素原料的物理結構,降低結溫度185-200℃、每100g干基原料加入0~3.3g晶度,包括球磨、剪切、擠壓等,其中最有效的是球Na2CO3時(shí)間5min、氧氣壓力0.3~1.2MPa),可磨,但由于能耗高而很少采用。溶解79%的半纖維素和49%的木質(zhì)素(生成羧22化學(xué)法酸)。此方法可處理城市垃圾、麥桿和玉米秸化學(xué)法:采用酸堿、有機溶劑或氧化劑等化學(xué)桿等草本纖維。試劑與纖維素原料進(jìn)行反應,以降解脫除原料中的2.3水熱化學(xué)法木質(zhì)素和半纖維素并溶解部分纖維素。常用的化學(xué)水熱化學(xué)法的主要特征是較高的溫度和水的存試劑有稀酸堿性試劑(氨、NaOH、石灰等)、氧化劑在,處理過(guò)程常伴隨化學(xué)反應。水熱處理時(shí)原料中(H2O2和氧氣等)或幾種試劑相結合。的乙?；纱姿?醋酸可進(jìn)一步進(jìn)行催化水解,因2.2.1稀酸水解預處理法而學(xué)汁了世恥為白動(dòng)水解法。稀酸水解預處理法主要是脫除原料中易于水解中國煤化工、蒸汽爆破(SE)、超的半纖維素和部分纖維素,以提高原料的酶可及度。臨界CNMH(常用的是SE法。SEyini等以米糠為原料,比較了稀酸、酶解和微生法原料用高壓飽和蒸汽處理一段時(shí)間后快速泄壓,物水解預處理的效果,其中,稀酸水解預處理法產(chǎn)生產(chǎn)生爆裂作用,從而減小原料尺寸和纖維素的結晶的還原糖量最多。將稀酸水解預處理法與物理法相度,高溫時(shí)有利于半纖維素的降解、木質(zhì)素的轉化第2期陳輝等.生物質(zhì)制燃料乙醇109使組分易于分離提高纖維素的酶可及度。由于 SE mmol/L)時(shí)纖維素降解生成的發(fā)酵抑制物的量增法中包含原料結構的物理破壞(爆裂)和化學(xué)反應,加,接種量為0.6g/L時(shí),乙醇相對產(chǎn)率可增至因此可用物理和化學(xué)參數表征SE過(guò)程中的變化,95%包括O與C比、H與C比、色度、元素分析、有機溶由于SE法在高溫、高壓條件下操作,故能耗較液含量、纖維素的結晶度及熱重分析等)。SE法高。陳輝將一種表面活性劑添加到SE法的預水與水熱處理法相比,SE法效果較好,纖維素的回收解過(guò)程中,纖維素可在相對較低的強度(185℃、5率達95%以上,且原料粒徑對酶解和發(fā)酵過(guò)程沒(méi)有min)下得到較好的處理效果。與不加表面活性劑相明顯的影響,可使用大粒徑原料3比,該法的酶解率提高了24%。將氨與表面活性劑一般的水熱處理法的效果較差,但在控制pH復配后,進(jìn)一步促進(jìn)了SE過(guò)程,酶解率達到93%。的條件下可促進(jìn)水解過(guò)程。玉米秸桿在溫度1602.4生物法℃、pH>4.0時(shí)進(jìn)行水熱處理,20min內可溶解生物法:利用可降解木質(zhì)素的微生物或酶選擇50%的纖維(其中80%為可溶性低聚糖,20%為單性地脫除原料中的木質(zhì)素?？山到饽举|(zhì)素的降解菌糖),降解損失率低于1%,且由于單糖量較少,故降株多為白腐菌(如 Basidiomycete, C Subvermispora,解產(chǎn)物的生成量也較少,低聚糖的量較多易于發(fā)酵 D Squalens, P Ostreatus, C Versicolor)1。生物法生成乙醇條件溫和,能耗低,無(wú)污染。與單純的醇解效果相SE法適用于農作物秸桿等草本纖維素原比,將醇解與白腐菌相結合的醇解效果可節約15%料,但只釆用SE法處理原料的苛刻度較高。在的電能?？山到饽举|(zhì)素的酶主要為酯酶255MPa3min時(shí),甘蔗渣達到最好的脫木質(zhì)素效 Anderson等用酯酶預處理草類(lèi)纖維,但酶處理液果如。SE法與稀酸水解預處理法結合可促進(jìn)草本中含有抑制發(fā)酵細菌生長(cháng)和乙醇生成的物質(zhì),脫除纖維素原料的水解,玉米秸桿在200℃、5min、質(zhì)量酶處理液中的酚類(lèi)物質(zhì)可提高乙醇的產(chǎn)率分數為2%H2SO4條件下,纖維素的轉化率比只采用SE法可提高4倍,且預處理水解液沒(méi)有明顯的毒素3發(fā)酵抑制物及其脫除抑制作用到。SE法與稀酸水解預處理法結合也適纖維素在預水解過(guò)程中發(fā)生了化學(xué)反應,生成用于木質(zhì)素含量高的原料,有效阻斷木質(zhì)素與纖維了對酶具有毒性或對發(fā)酵過(guò)程具有抑制作用的物素酶間的作用,使水解產(chǎn)率提高5%~20%,纖維素質(zhì)。這些物質(zhì)可采用物理、生物或化學(xué)手段脫除,也酶的利用率提高50%32可在控制水解過(guò)程中減少抑制物和毒素的生成。兩步稀硫酸浸漬SE法:首先在低強度(180℃、 Martin等(釆用SE法預處理土豆桿,水解液中抑10min、質(zhì)量分數為0.5%H2SO4)下用SE法水解制物的含量較低,但隨處理時(shí)間的延長(cháng),抑制物的含提取半纖維素,然后在高強度(200℃、2min質(zhì)量分量增加。近年來(lái)構建了許多耐受抑制物的工程菌,數為2%H2SO4)下用SE法水解部分纖維素并使纖可省略脫除過(guò)程降低成本。維素更易于酶解(此時(shí)乙醇的產(chǎn)率可達到最大3.1脫除方法值)。 Soderstrom等進(jìn)一步將稀硫酸浸漬SE發(fā)酵抑制劑的種類(lèi)和數量與生物質(zhì)的種類(lèi)和預法與SO2注入SE法相結合,第一步在低強度(180處理方法有關(guān),其抑制機理也各有不同。 Helle℃、10min、質(zhì)量分數為0.5%H2SO4)下預處理云等研究了抑制物對木糖發(fā)酵產(chǎn)生乙醇工程菌S杉第二步在高強度(210℃、5min質(zhì)量分數為3% Cerevisiae的影響醋酸、氨糠醛對工程菌和其母株SO2)下預處理云杉,發(fā)酵乙醇的產(chǎn)率可達到理論值均具有抑制作用。 Klink等4研究了纖維素高溫的66%,間接發(fā)酵(SHF)的乙醇產(chǎn)率可達到理論值物理預處理的水解產(chǎn)物,其中糖降解產(chǎn)物呋喃和木的71%質(zhì)素降解產(chǎn)物單酚是最主要的發(fā)酵抑制物,會(huì )抑制氨纖維爆破預處理(AFEX)法與SE法相似,只酒精什旨和的速率,但不會(huì )影響燃是前者采用液氨為介質(zhì)。AFEX法可提高纖維素的料乙中國煤化工研究了SE法預處酶解纖維素轉化率從未處理的16%增至93%,乙理過(guò)CNMHG醋酸、乙酰丙酸、糠醇產(chǎn)率可提高22~2.5倍3.3醛、羥甲基糠醛(HMF)、丁香醛、羥基苯甲醛、香蘭Fenton試劑(H2O2和Fe2)的氧化反應也能加醛)對纖維素酶活性的影響實(shí)驗結果表明,甲酸可強SE法的預水解過(guò)程,但在使用高濃度H2O2(50使酶失活,香蘭醛可部分影響酶的活性,其他毒素對石油化工PETROCHEMICAL TECHNOLOGY2007年第36卷酶活性的影響不大。ZM4AcR細胞內的pH和能量狀態(tài)基本上不受醋酸抑制物的脫除可采用吸附法、離子交換鈉的影響,因而具有很高的耐受性法64、萃取法、生物法、過(guò)量堿中和法等?？啡┖虷MF是生物質(zhì)降解產(chǎn)物中最主要的兩吸附法需使用選擇性吸附劑,較好的吸附劑有種發(fā)酵抑制物,高耐受毒素的 s Cerevisiae和P木炭、聚合樹(shù)脂等。weli等研究了苯乙烯基 Stipitis可將HMF轉化為2,5-雙-(羥甲基呋喃),聚合物XAD-4吸附劑對糠醛脫除效果的影響,實(shí)糠醛轉化為糠醇5驗結果表明,糠醛的質(zhì)量濃度可從1~5g/L降至0.01g/L。吸附劑可用乙醇脫附和再生。4纖維素酶和發(fā)酵生產(chǎn)乙醇的微生物生物法需使用可代謝抑制物或毒素的微生物或4.1纖維素酶酶。 C Ligniaria NRRL30616可脫除玉米秸桿稀酸纖維素酶是混合酶,主要包括內切葡聚糖酶、外水解液中的發(fā)酵抑制物。 T Reesi能降解蒸汽預切葡聚糖酶(CBH)和β-葡萄糖苷酶,三者協(xié)同作處理柳樹(shù)過(guò)程中半纖維素水解所產(chǎn)生的抑制物,從用。纖維素的酶分子普遍具有類(lèi)似的結構,由催化而將燃料乙醇的產(chǎn)量提高4倍,產(chǎn)率提高3倍。結構域、連接橋和纖維素結合結構域(CBD)3部分生物酶對底物具有專(zhuān)一性,因而應用較少。如漆酶組成。最近的研究表明,這些區域的功能不是惟可脫除單酚和苯酚,但對大分子苯系化合物僅具有的,CBD區域對多聚糖結構也具有破壞作用,因而降低相對分子質(zhì)量的作用,不能將其轉化為不具發(fā)也可促進(jìn)水解的進(jìn)行。Vaje- Kolstad等3的研酵抑制性的物質(zhì)究結果表明,CBD中的少量非催化性蛋白(CBP21)過(guò)量堿中和法是采用堿性物質(zhì)脫除水解液中抑對殼質(zhì)的降解具有決定作用。CBP21會(huì )改變底物制物的方法,對于稀酸水解液,Ca(OH)2過(guò)量堿中的結構,促進(jìn)纖維素水解的進(jìn)行。纖維素酶的CBD和是最常用的脫除方法。C核磁共振分析結果顯參與了酶與木質(zhì)素的結合,但無(wú)CBD的纖維素酶仍示{3),過(guò)量堿中和法脫除的主要是脂肪酸和芳香性對木質(zhì)素表現出親合力酸或酯及其他芳香和脂肪化合物,酮和醛官能團沒(méi)可發(fā)酵產(chǎn)生纖維素酶的微生物有木霉有明顯的變化。 O'Brien等2比較了Ca(OH)2過(guò)( Trichoderma)、曲霉( Aspergillus)、青霉( Penicillum量堿中和法和強堿陰離子中和(AEN)法對玉米纖和 Humicola)、鐮刀菌( Fusariun)、鏈霉菌屬維稀酸水解液中抑制物的脫除效果,實(shí)驗結果表明,( Streptomyces)、芽孢桿菌屬( Bacillus)、纖維單胞菌AEN法的脫除率更高。 Cantarella等比較了不同屬( Cellulomonas)等。纖維素酶的生產(chǎn)有固態(tài)發(fā)酵抑制物脫除方法,實(shí)驗結果表明,Ca(OH)2過(guò)量堿法和液態(tài)發(fā)酵法。一般采用液態(tài)發(fā)酵法,纖維素中和法效果最好,水解液脫除抑制物后發(fā)酵生產(chǎn)乙酶較易于提取,其生產(chǎn)成本可隨著(zhù)規模的擴大而醇的產(chǎn)率可達到理論產(chǎn)率的92%。降低,但由于纖維素酶的生產(chǎn)需固體纖維素的誘由于水解液中抑制物主要在發(fā)酵初期起作用,導,生產(chǎn)周期長(cháng),效率低,且纖維素酶解耗酶量大,隨發(fā)酵的進(jìn)行,細胞繁殖彌補了抑制物造成的細胞因而較高的纖維素酶生產(chǎn)成本大幅度降低了生物死亡,維持了一定的細胞濃度水平,因而可釆用高接質(zhì)制燃料乙醇的經(jīng)濟性。近年來(lái)里氏木霉纖維素種量消除發(fā)酵初期抑制物的作用。此外,還可采酶的生產(chǎn)成本大幅度降低(不到原來(lái)生產(chǎn)成本的用固定化酵母法。酵母膠囊化后可降低失活速率,1/10),乙醇酶的生產(chǎn)成本降至0.026~0.053但由于水解液的毒性太大膠囊化的酵母也會(huì )逐漸美元/L6,因而生物質(zhì)制燃料乙醇的經(jīng)濟性可望喪失活性。大幅度提高。3.2耐毒微生物為改善纖維素酶的性能、降低纖維素酶用量,在近年來(lái),由于對發(fā)酵抑制物毒理認識水平的提產(chǎn)酶菌株的改良和構建方面展開(kāi)了許多研究。文獻高,采用馴化或基因改造等生物技術(shù)改良菌種,可提[62高某些發(fā)酵微生物對抑制物的耐受性。中國煤化工和誘變易錯聚合酶鏈結合,產(chǎn)生了變種TKim等6研究了抑制物對 Z Mobilis zMμ及其ReoCNMHG7A。將此變異基因耐毒素變異菌株zM4/AcR發(fā)酵生產(chǎn)乙醇的抑制作在 S Serevisiae中表達并篩選以提髙熱穩定性和高用,3P核磁共振分析結果顯示,細胞內酸化和能量溫活性。其中一個(gè)變種被認定在TRei中代替了激勵是醋酸鹽顯示抑制性的兩種主要機理。原生的cel7A。重整菌株酶對預處理玉米秸桿的第2期陳輝等.生物質(zhì)制燃料乙醇111水解效果超過(guò)親本。 Bower等將細菌A60542), s Cerevisiae EF10140, s CerevisiaeCellulolyticus的內切葡聚糖酶GH5A基因融合到真424A(431, s Cerevisiae tmB34001等;構建了可菌 T Reesi的纖維二糖水解酶CBH上,融合蛋白在發(fā)酵阿拉伯糖的工程菌有 Becker構建的ST Reesi中表達得到的菌株酶對預處理玉米秸桿的 Cerevisiae工程菌;構建了具有特定耐受性的發(fā)酵木糖化水解更有效糖生產(chǎn)乙醇的工程菌有:有氧發(fā)酵可耐受稀酸水解轉基因植物纖維素酶是一個(gè)較新的研究方向,液的 M Indicus1,耐高溫并具有較高乙醇耐受性已將 A Cellulolytic的內切葡聚糖酶和 T Reesi的纖的F-71m,可有氧發(fā)酵的 C Opuntiae 1605N),耐維二糖水解酶基因表達在馬鈴薯和西紅柿中。這些受抑制物的 T Mathranii a3M34等。植物纖維素酶與微生物纖維素酶相似,但生產(chǎn)成本4.3可直接利用纖維素原料的微生物大幅度降低。以?xún)惹衅暇厶菫槔?每千克酶的成本纖維素原料的水解糖化和發(fā)酵產(chǎn)生乙醇可在同由原來(lái)的5美元降至1.40美元。Hood等6將株微生物中完成。如可利用纖維素直接生產(chǎn)乙醇多糖降解酶表達在農作物的種子組織中,轉基因作的 C Thermocellum),可發(fā)酵微晶纖維素和多種糖物的種子組織可作為酶源,從而可降低燃料乙醇的生產(chǎn)乙醇的 Trichoderma A10,可發(fā)酵纖維素和生產(chǎn)成本。己糖生產(chǎn)乙醇的 K Marxianus,在產(chǎn)醇酵母細胞4.2可代謝戊糖生產(chǎn)乙醇的基因工程菌表面共表達3種纖維素酶組分可構建發(fā)酵無(wú)定形纖半纖維素在生物質(zhì)中占有相當大的比例,其水維素的工程菌8,在可發(fā)酵木糖的釀酒酵母細胞表解產(chǎn)物主要是木糖,以農作物秸桿和草為原料時(shí)水面共表達兩種木聚糖分解酶(半纖維素酶和半纖維解產(chǎn)物中還有一定量的阿拉伯糖(可占戊糖質(zhì)量的素二糖酶)可構建發(fā)酵木聚糖的工程酵母(。10%~20%),故發(fā)酵戊糖生產(chǎn)乙醇的效率也是決4.4提高特定耐受性的微生物定該過(guò)程經(jīng)濟性的重要因素根據過(guò)程需要,在耐熱性、有氧發(fā)酵、抑制物耐葡萄糖的代謝途徑與木糖不同,且往往對木糖受能力等方面也進(jìn)行了工程菌的構建?；谕瑫r(shí)糖的代謝產(chǎn)生抑制作用。 C lusitaniae可發(fā)酵木質(zhì)纖化和發(fā)酵法的需要,化學(xué)誘變 K Marxianus DER-維素水解液中的葡萄糖生成乙醇,在水解液中無(wú)葡26得到了耐熱菌工程菌CECT10875M。有氧發(fā)萄糖時(shí)可同時(shí)發(fā)酵纖維二糖和木糖生成乙醇的。S酵的變異酵母可在發(fā)酵的同時(shí)細胞生長(cháng)并保持活Cerevisiae tmB3001, CPB CR1, CPB CRA2也是先性,生產(chǎn)效率高、乙醇的產(chǎn)率高;釀酒酵母中表達發(fā)酵葡萄糖后再利用木糖(6。利用分子生物學(xué)可ADH6可提高乙醇的生產(chǎn)力,毒素的耐受力強構建發(fā)酵戊糖和己糖生產(chǎn)乙醇的基因工程菌。常用5水解和發(fā)酵工藝的釀酒酵母 S Cerevisiae不能發(fā)酵木糖生成乙醇,但能利用木酮糖,可將戊糖代謝為木酮糖的關(guān)鍵酶引發(fā)酵法制乙醇的生產(chǎn)工藝有3種:分別水解發(fā)入到酵母中。 e Coli, P Stipitis, Z Mobil可利用酵SHF(法)、同時(shí)糖化發(fā)酵(SSF)法、直接發(fā)酵法。木糖發(fā)酵生產(chǎn)乙醇,但需脫除發(fā)酵液中的抑制物,且5.1SHF法乙醇產(chǎn)率較低,可在這類(lèi)菌株中引入高效產(chǎn)生乙醇SHF法:先水解糖化再發(fā)酵的二步發(fā)酵法。纖的關(guān)鍵酶。ECol厭氧發(fā)酵產(chǎn)物中的乙醇含量低,維素原料的水解糖化方法主要有稀酸水解、濃pH控制在6.0~8.0,其基因改造主要是適應稀酸酸水解、酶解。濃酸水解目前采用的不多,主要有日水解液條件,并控制代謝產(chǎn)物向乙醇方向進(jìn)行。P本JBC公司和美國 Masada OxyNol LLC公司的sipi中含有代謝木糖的木糖還原酶和木糖醇脫氫 Masada CES OXyNol過(guò)程。酶解法需進(jìn)行原料的酶,但對木糖的代謝卻受葡萄糖的抑制,對乙醇和纖預處理,使用纖維素酶催化水解。維素水解抑制物的耐受性差,且需控制微好氧條件發(fā)酵菌種有時(shí)不能同時(shí)利用水解液中的混合發(fā)酵。 Z Mobilis工程菌發(fā)酵葡萄糖和木糖的動(dòng)力糖,或對抑制物敏感.因而可對水解液進(jìn)行分離,或學(xué)研究表明,醋酸能強烈抑制發(fā)酵過(guò)程6,重整菌采中國煤化工或采用一步發(fā)酵法。Z Mobilis8b較能耐受醋酸,pH=6時(shí)乙醇產(chǎn)率可達CNMH或可發(fā)酵混合糖的菌到85%6種?？刹捎蒙V分離法分離水解液中的糖,吸附劑近年來(lái),構建了可發(fā)酵木糖的工程菌有:ECol可采用 Dowex99和聚乙烯吡啶(PVP)等。PVPKoI0, P Stipitis CBS57731, P Stipitis CBS模擬移動(dòng)床可有效分離玉米秸桿水解液中的糖(阿石油化工112·PETROCHEMICAL TECHNOLOGY2007年第36卷拉伯糖、甘露糖、木糖、半乳糖、葡萄糖、纖維二糖)汽化工藝結合了膜分離和蒸發(fā)過(guò)程,可大幅度降低和雜質(zhì)(硫酸、醋酸、糠醛、羧甲基糠醛)明。能耗,提高生物質(zhì)制燃料乙醇的經(jīng)濟性,是最有應用5.2SSF法前景的分離技術(shù)。膜材料可采用殼聚糖衍生SSF法將酶解和發(fā)酵放在同一個(gè)反應器中進(jìn)物、聚二甲基硅氧烷m等。其他脫水技術(shù)還行,可克服酶解過(guò)程中的反饋抑制作用,簡(jiǎn)化了生產(chǎn)有分子篩吸附法等,常需與蒸餾過(guò)程相結合。工藝,但存在的主要問(wèn)題是酶解和發(fā)酵溫度條件的不一致性。酶解適宜的溫度約為50℃,而發(fā)酵的控7生物質(zhì)制燃料乙醇的項目制溫度是37~40℃。解決的方法:一是采用非等溫較大的生物質(zhì)制燃料乙醇的項目有美國的可再過(guò)程,二是利用耐熱酵母的等溫過(guò)程。等溫過(guò)程一生能源實(shí)驗室(NREL)開(kāi)發(fā)的NREL過(guò)程、加拿大般比非等溫過(guò)程的乙醇產(chǎn)率高。近年來(lái)耐熱酵的loge過(guò)程、印度的 T Delhi過(guò)程等。母構建取得了很大進(jìn)展,等溫SSF法存在的問(wèn)題已NREL過(guò)程是專(zhuān)門(mén)立項的“生物乙醇”項目解決。( DOE Bioethanol Program)。NREL過(guò)程(1999)6SSF法與SHF法相比,雖SHF法乙醇的產(chǎn)率是以玉米秸桿為原料,通過(guò)稀酸預處理(190℃、10高,但SSF法耗時(shí)短,燃料乙醇的產(chǎn)量高明。 SSF min),同時(shí)糖化和共發(fā)酵(木糖和葡萄糖)生產(chǎn)燃料法有連續或半連續工藝,半連續的SSF法可減少酶乙醇。預處理產(chǎn)物快速閃蒸分離殘留在液相中的的用量醋酸通過(guò)連續的離子交換去除,漿料以石膏過(guò)量堿5.3直接發(fā)酵法中和脫毒。預水解液和漿料再進(jìn)行混合生產(chǎn)燃料乙直接發(fā)酵法是直接發(fā)酵纖維素生產(chǎn)乙醇,這種醇。發(fā)酵菌為可同時(shí)利用木糖和葡萄糖的重整Z方法設備簡(jiǎn)單,但也存在一些問(wèn)題。如纖維素發(fā)酵 Mobili該方法的發(fā)酵溫度較低(30℃),發(fā)酵時(shí)間速率慢、容積生產(chǎn)力低;發(fā)酵產(chǎn)物中含有乙酸、乳酸長(cháng)達7d。乙醇采用蒸餾和分子篩吸附提純。蒸餾等有機酸和氫氣等,乙醇產(chǎn)率低。常用的解決方法塔底出料為固體木質(zhì)素/細胞,可燃燒生產(chǎn)蒸汽,是與不分解纖維素的嗜熱菌混菌培養,利用游離單蒸汽用來(lái)發(fā)電,塔底出料液體蒸發(fā)濃縮回收水。糖產(chǎn)生乙醇。如耐熱 clostridia可在厭氧條件下用這一過(guò)程每升的燃料乙醇生產(chǎn)成本約為0.395美于混菌培養并提高燃料乙醇的產(chǎn)量。元,若提高預處理過(guò)程效率,提高發(fā)酵溫度54發(fā)酵過(guò)程的促進(jìn)(55℃),構建可利用多種糖的產(chǎn)醇重組菌,提高發(fā)酵過(guò)程可用磁場(chǎng)、添加表面活性劑等促進(jìn)方纖維素酶的生產(chǎn)能力和酶活性,每升燃料乙醇的法提高乙醇產(chǎn)率和產(chǎn)量。強磁場(chǎng)不會(huì )改變微生物細成本可降至0.248美元。最近有報道稱(chēng)NREL的胞的宏觀(guān)特征,僅影響生物量和燃料乙醇的產(chǎn)合作伙伴 Novozymes公司降低了纖維素酶的生產(chǎn)量,可使生物量提高2.5倍,乙醇濃度提高3.4成本,每升燃料乙醇消耗纖維素酶的成本由最初倍叨。在SSF過(guò)程中加入吐溫20(約25gL),反的1.32美元降至0.079美元,美國 Genencor公司應速率加快,乙醇產(chǎn)率提高了8%,同等乙醇產(chǎn)率下甚至降至0.026~0.053美元。生物法低成本酶的用量可減少50%,SSF過(guò)程結束時(shí)酶的活性有生產(chǎn)燃料乙醇面臨突破。所提高。logen公司是加拿大工業(yè)纖維素酶和半纖維素通過(guò)完善酶解發(fā)酵工藝,也可提高乙醇產(chǎn)率,降酶的主要生產(chǎn)商,2003年建造了40td的纖維素制低生產(chǎn)成本。濃縮SE預水解液時(shí),抑制物的濃度燃料乙醇的裝置(并與其酶的生產(chǎn)相結合)。此后也同時(shí)增加,降低了可發(fā)酵性。在濃縮物中添加富 logen公司和加拿大石油公司合作投產(chǎn)了世界上最含葡萄糖的纖維素水解液,可顯著(zhù)改善發(fā)酵效大的以纖維素廢料為原料生產(chǎn)燃料乙醇的工業(yè)裝果∞。纖維素酶和纖維二糖酶的成本高,可采用超置,可使12-15k/a谷物秸桿轉化為(3~4)×10濾回收。L/a燃料乙醉,預計采用新技術(shù)后生產(chǎn)費用可降至6乙醇濃縮提純工藝中國煤化工將纖維素原料進(jìn)行稀酸CNMH(纖維素酶水解糖從生物質(zhì)發(fā)酵液中提純乙醇傳統的工藝是采化,最后進(jìn)行SHF發(fā)酵生產(chǎn)乙醇。預處理產(chǎn)物用蒸餾的方法,由于乙醇與水可形成共沸物,因此需用堿調節pH=5,加入纖維素酶發(fā)酵原液在50℃水萃取精餾、恒沸精餾以制取無(wú)水乙醇,能耗高。滲透解5~7d。水解固相過(guò)濾分離,濾液與水解液以酒第2期陳輝等.生物質(zhì)制燃料乙醇113精酵母發(fā)酵葡萄糖生產(chǎn)乙醇,蒸餾提純乙醇。木半纖維素可發(fā)酵生產(chǎn)乙醇、木糖醇、丁二醇等2質(zhì)素用于燃燒提供熱量和蒸汽。 Iogen過(guò)程最大木質(zhì)素可直接燃燒轉化為能量,或熱解生產(chǎn)合成氣的優(yōu)勢在于其纖維素酶的現場(chǎng)利用,以原液方式燃料油,或用于生產(chǎn)木質(zhì)素磺酸鹽等化學(xué)品;高純度加入,省去了酶的制備費用,且一部分水解液可用的纖維素可降低纖維素酶的用量,提高葡萄糖產(chǎn)率。于酶的生陳洪章等21將SE法處理的麥草用水抽提出其中IIT Delhi過(guò)程6以稻草為原料,預處理采用自的半纖維素,再用乙醇萃取木質(zhì)素后得到纖維素,纖動(dòng)水解和乙醇抽提脫木質(zhì)素兩步法。自動(dòng)水解過(guò)程維素的酶解率可達到90%以上。Lgno過(guò)程以乙醇中約有70%的半纖維素水解,水解液中含以木糖為為溶劑從木材中分離木質(zhì)素和半纖維素以獲得具有主的多種糖,此水解液用于培養 C Utilis(用作動(dòng)物酶可及度高的纖維素,乙醇和水可循環(huán)利飼料)。自動(dòng)水解的固體殘渣在催化劑的存在下用13,1。 Pure vision過(guò)程是將木質(zhì)纖維素原(170℃30min),用體積分數50%的乙醇抽提脫木料用水、稀堿、堿性氧化劑在較高溫度下采用串聯(lián)、質(zhì)素。兩步法預處理后的纖維素(76%)進(jìn)行SSF。逆流的方法洗滌,然后在獨立的反應器中連續進(jìn)行SSF過(guò)程采用TRe纖維素酶 A Wentii B-葡萄SE,其液相組分中主要含有木質(zhì)素、半纖維素和其糖苷酶、 C Acidothermophilum為發(fā)酵菌種。SSF過(guò)程他抽提物,固相組分主要是高反應活性的高純度纖采用間歇加料,伴有程序真空回收過(guò)程。由于溶劑維素。預處理過(guò)程所需能耗高,燃料乙醇的生產(chǎn)成本約為9結語(yǔ)0.544美元/L。近年來(lái),生物質(zhì)制燃料乙醇的研究和應用取得8生物質(zhì)全利用整體工程了很大的進(jìn)展,燃料乙醇的生產(chǎn)成本大幅度降低,但規模經(jīng)濟可降低生產(chǎn)成本,但植物纖維素原料燃料乙醇的生產(chǎn)僅是整個(gè)生物質(zhì)加工工程的一部的運輸往往限制其規模的擴大。事實(shí)上,由生分,從可再生的生物質(zhì)獲取多種產(chǎn)品和能量將是未物質(zhì)制燃料乙醇的能耗高于乙醇燃燒生成的能量,來(lái)不可阻擋的發(fā)展方向。近期的發(fā)展方向為:如以玉米粒為原料的能耗比產(chǎn)能高29%,以軟枝草(1)繼續降低纖維素原料制燃料乙醇的成本。為原料的能耗比產(chǎn)能高50%以上,以木材為原料的目前,纖維素原料制燃料乙醇的成本仍然很高,其制能耗更高(為57%以上)m。因而,單一乙醇產(chǎn)品約因素主要是預處理過(guò)程和纖維素酶因而有待在的經(jīng)濟性差 Hamelinck的技術(shù)經(jīng)濟評估認為,基于稀技術(shù)和菌種上進(jìn)一步突破。酸水解工藝,生產(chǎn)效率為35%,結合木質(zhì)素發(fā)電后生(2)擴大乙醇的利用范圍。由于乙醇易于反應產(chǎn)效率可達到60%。改進(jìn)預處理和生物技術(shù)后,生生產(chǎn)乙烯因而目前95%來(lái)自石油的合成聚合物可產(chǎn)效率可達48%和68%。因而生物質(zhì)制燃料乙從生物質(zhì)資源中獲得0醇生產(chǎn)過(guò)程應形成多元化產(chǎn)品或產(chǎn)能的整體工程,提(3)提高生物質(zhì)整體化工程的技術(shù)經(jīng)濟性。未高整體經(jīng)濟性,以降低燃料乙醇的生產(chǎn)成本。來(lái)的生物質(zhì)加工廠(chǎng)的經(jīng)濟效益來(lái)自多種產(chǎn)品而不是整體化工程是生物質(zhì)和能量全利用的過(guò)僅依靠一種產(chǎn)品。正如石油煉廠(chǎng)的多模式一樣,生程:一要結合多種加工方法,如發(fā)酵、蒸汽重整、氣物質(zhì)加工廠(chǎng)也應面向市場(chǎng),因地制宜,具有足夠的靈化、液化、熱解、水解、快速裂解1電解;二要活性。充分利用生物質(zhì),形成多元化產(chǎn)品。日本充分利用發(fā)酵廢渣,進(jìn)行水熱催化氣化生成乙醇,乙醇產(chǎn)率由25%增至30%~36%l2 wilkening等利用釜餾物清液生產(chǎn)甲烷氣。多元化產(chǎn)品可以燃料(如錢(qián)伯章.世界乙醇產(chǎn)業(yè)開(kāi)始步入黃金發(fā)展期.中國石化,2006(4):26-27CH4、氫氣、合成氣、乙醇生物油等)為中心,還2張及前景.中國能源,2006可副產(chǎn)天然纖維、甜味劑、單細胞蛋白、乳酸、微中國煤化工生物酶16、活性炭、優(yōu)質(zhì)蛋白飼料8、土壤改CNMHGConstruction Waste.JJap良劑9、黏合劑1等ns Energy,2005,84(4):290-292Okuda N. 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