溶剂脱毒汽爆玉米秸秆对酶解以及发酵的影响

以脱毒对酶解及发酵的影响为研究对象,以酶解还原糖得率及发酵乙醇质量浓度为指标,采用溶剂萃取的方法对无催化汽爆玉米秸秆进行萃取脱毒。结果表明,酶解还原糖得率随着萃取剂及萃取方式介于34.85%和89.7%,酶解还原糖得率和发酵乙醇质量浓度与脱毒有机溶剂的沸点高度负相关,表明有机溶剂的残留是导致酶失活的主要原因。而对于所考察的溶剂,乙醇产率为0.47～0.49 g乙醇/g还原糖,表明有机溶剂残留对乙醇发酵并无显著影响。采用乙醚和丙酮的组合萃取,乙醇最高产率可以达到理论值的96.1%。     

玉米秸秆氨化汽爆处理及其固态发酵

在加氨条件下对玉米秸秆进行了汽爆处理(简称氨化汽爆)和固态发酵. 结果表明: 氨化汽爆同样可使秸秆中的半纤维素降解,并使玉米秸秆的酶解率提高到42.97%, 同时可使秸秆的有机氮含量提高1.27倍. 利用氨化汽爆秸秆进行固态发酵,可提高蛋白含量到23.45%,比不加氨汽爆的玉米秸秆提高了1倍. 而加过氧化氢的氨化汽爆不利于微生物发酵.     

β-葡萄糖苷酶高温同步糖化发酵产乙醇应用研究

利用Trichoderma sp.W2所产的嗜温耐乙醇β-葡萄糖苷酶及耐高温酵母Kluyveromyces marxianus NCYC 587,以气爆秸秆为原料进行高温同步糖化发酵。研究结果表明:在42℃条件下,接种体积分数10%,底物质量分数15%,发酵pH值为4.8,β-葡萄糖苷酶添加量为30 U/g底物条件下发酵效果最好。NCYC 587能迅速利用预水解产生的葡萄糖发酵并积累乙醇,同时能利用部分木糖,但在发酵后期,葡萄糖利用完全后会代谢利用一定量的乙醇,致使发酵过程中乙醇质量浓度始终维持在一个相对较低的水平。乙醇最高质量浓度达到20.56 g/L,乙醇产率达80.64%。添加嗜温耐乙醇β-葡萄糖苷酶于高温同步糖化发酵能有效解决纤维素酶解发酵过程终端产物抑制的难题。     

汽爆玉米秸秆发酵丁醇的酶解工艺优化

为了提高秸秆酶解后的还原糖浓度和酶解液发酵后的丁醇产量,研究了不同因素对汽爆玉米秸秆酶解的影响,优化汽爆玉米秸秆秸秆发酵丁醇的酶解工艺。结果表明汽爆玉米秸秆的最佳酶解工艺为:反应温度50℃、原始底物浓度15%(wt)、酶用量60 IU·(g底物)-1、酶解时间48 h、搅拌器转速100 r·min-1。通过考察分步加料方式,三次加料方式最高表观黏度和反应后期表观黏度都低于两次加料的方式,而其糖浓度则略高于两次加料方式。当底物从原始浓度15%(wt)分三次加入到25%(wt)时,还原糖浓度、丁醇产量分别为89.23、9.82 mg·mL-1,分别增加了58.63%、44.20%。     

氢氧化钠预处理对甘蔗渣酶解和发酵性能的影响

采用氢氧化钠预处理甘蔗渣,通过单因素和正交试验考察了不同预处理条件对甘蔗渣酶解和发酵性能的影响,并进一步分析了比表面积和木质素含量对酶解性能的影响。结果表明：预处理温度、氢氧化钠质量分数及预处理时间对酶解和发酵效率影响较为显著,最佳的预处理条件为：温度85℃、时间11 h、NaOH质量分数4.5%,在此优化条件下预处理的甘蔗渣,含纤维素56.46%,与原料相比提高了46.16%;半纤维素20.30%、Klason木质素5.79%,与原料相比分别降低了15.77%和72.87%,酶解36 h的还原糖得率为0.69 g/g（以甘蔗渣质量计）。经过氢氧化钠预处理后的甘蔗渣比表面积显著增加（由原料的0.07 m²/g最大可增加到1.07 m²/g）,木质素显著降低,有利于提高酶解和发酵效率。当比表面积超过0.30 m²/g时,酶解初始速率和酶解效率达到平衡;当木质素低于11%时,酶解效率达到平衡。     

过氧化氢-乙酸联合预处理对甘蔗渣酶解和发酵的影响

采用过氧化氢-乙酸(HPAC)对甘蔗渣(SCB)进行了联合预处理。以预处理后的甘蔗渣为原料, 先进行酶水解, 然后将水解液进行乙醇发酵, 探讨预处理对甘蔗渣酶解和发酵的影响。实验结果表明: 20 g甘蔗渣, 加入150 mL过氧化氢水溶液(75 mL过氧化氢(30%)和75 mL水)和150 mL乙酸(99%), 硫酸用量为过氧化氢-乙酸溶液体积的0.5%, 在70 ℃反应2 h时, HPAC预处理脱除了88.85%的木质素, 并使90.10%的纤维素保留在底物中。底物(HPAC/70-SCB-0.5)的酶可及度是80.30 mg/g, 与相同条件下单独过氧化氢预处理(HP/70-SCB)和单独乙酸预处理(AC/70-SCB)相比, 分别增加了38.26%和31.08%, 甘蔗渣木质素的表面覆盖率从原料的0.66降低至0.22。酶解上清液在酶用量为5 FPIU/g(以底物计)条件下水解后, 葡萄糖得率是87.63%, 分别是HP/70-SCB和AC/70-SCB的6.89和20.62倍, 发酵产乙醇质量浓度是7.57 g/L, 分别是HP/70-SCB和AC/70-SCB的7.65和22.94倍。     

汽爆秸秆膜循环酶解耦合丙酮丁醇发酵

利用新型的汽爆玉米秸秆膜循环酶解耦合发酵系统进行了丙酮丁醇发酵的研究,并对使用该系统所导致的丙酮丁醇梭菌(Clostridium acetobutylicum AS1.132)代谢的变化进行了讨论. 在稀释率为0.075 h-1的条件下,丁醇的产量为0.14 g/g (纤维素+半纤维素),最大丁醇产率达到0.31 g/(L×h),溶剂组成为丁醇:丙酮:乙醇65.3:24.3:10.4(体积比),纤维素和半纤维素的转化率分别为72%和80%,使用单位纤维素酶所产生的丁醇量为3.9 mg/IU,是分步水解批次发酵的1.5倍. 利用该系统使酶解和发酵分别在各自最适的条件下同时连续进行,减少了纤维素酶的用量,有效地解除了酶解产物对纤维素酶的抑制作用,并减轻了溶剂产物尤其是丁醇对微生物活性的影响,延长了发酵周期.     

乙酸预浸对玉米秸秆蒸汽爆破预处理的影响

对经乙酸预浸汽爆预处理的玉米秸秆进行了组分含量和抑制物分析,并研究了玉米秸秆预处理后的酶水解性和同步糖化发酵。与未经乙酸预浸相比,乙酸预浸玉米秸秆能在相对低温下进行汽爆预处理,在提高半纤维素水解程度的同时,并不会明显增加糠醛等发酵抑制物。酶水解实验表明,玉米秸秆经乙酸预浸,再以 200 ℃ 进行汽爆后的酶水解效果较好,每克原料可获得 284 mg 葡萄糖,提高了 10.2 %,为理论值的 76.8 %;乙酸预浸玉米秸秆经过 96 h 同步糖化发酵,获得了 22.5 g/L 的乙醇浓度,为理论值的 72 %;相比未经乙酸预浸的玉米秸秆,提高了11.9个百分点。     

木糖利用融合子D2的制备与乙醇发酵特性

以树干毕赤酵母（Pichia stipitis）1960（Ps1960）与酿酒酵母（Saccharomyces cerevisiae）AADY（ScAADY）为亲本菌株,采用双亲灭活原生质体技术制备木糖利用融合子,并对其制备条件进行了优化。优化后的原生质体制备条件为Ps1960采用2%蜗牛酶和1%纤维素酶在28℃酶解45 min,20W紫外灯距离10 cm照射3 min灭活;ScAADY采用1.5%蜗牛酶和1%纤维素酶28℃酶解50 min,55℃水浴50 min灭活;均采用0.6 mol/L山梨醇为渗透压稳定剂。在该条件下,共得到22株融合子。通过测定各融合子在不同培养基条件下的生物量来评价其木糖代谢和乙醇耐受能力,最终获得能利用木糖高效发酵产乙醇、遗传性状稳定的融合子D2,并进行乙醇发酵条件优化。结果表明,在混合糖质量分数8%、木糖和葡萄糖质量比6：1、5%接种量、30℃、160 r/min、培养72 h条件下,融合子D2发酵产乙醇的产量为40.58 g/L。     

响应面法优化水/醇处理后汽爆玉米秸秆酶解

为了提高水/醇处理后汽爆玉米秸秆的酶解还原糖产率,对其酶解条件进行了优化。通过响应面优化法确定了底物质量浓度为53.28 g/L,纤维素酶用量为53.32 FPU/g,酶解时间为60.45 h时,还原糖产率可达672.36mg/g,与秸秆物料及汽爆后物料相比,酶解还原糖产率分别提高了170.46%和28.97%。化学组分及结构形貌分析表明,汽爆水/醇处理后物料纤维素含量显著增加,物料相对结晶度增高,其结构更有利于纤维素酶分子的吸附。     

蔗渣闪爆处理及其黄原酸化物的制备和应用

采用热蒸汽适度闪爆及稀碱洗涤等预处理技术对蔗渣进行纯化和活化,利用处理后的蔗渣纤维素合成纤维素基黄原酸酯,对其在水处理中的应用进行了研究。研究优化了闪爆处理的工艺条件,并采用IR、SEM和化学分析技术对闪爆前后蔗渣纤维的形态、结构、a-纤维素的含量进行了分析,对处理前后的蔗渣纤维的碱化和黄原酸化合成条件进行了优化。结果表明,闪爆预处理技术是一种便宜、迅速、无污染的技术,蔗渣纤维素基黄原酸酯对含金属离子的污水有良好的处理效果。     

稀硫酸预浸渍对玉米秸秆蒸汽爆破预处理的影响

为提高纤维素乙醇生产中传统蒸汽爆破预处理的效果,以稀硫酸（质量浓度0．2％）对玉米秸秆进行预浸渍,再于190～210℃对其进行汽爆预处理。结果表明：稀硫酸预浸渍有利于增强汽爆过程中半纤维素的水解程度,并能有效减少乙酸的生成;200℃预处理玉米秸秆经过96h同步糖化发酵,最终乙醇浓度为22．5g·L-1,为理论值的76％,较预浸渍前（19．0g·L-1）明显提高。稀硫酸预浸渍能够增强玉米秸秆的汽爆预处理效果。     

Enhanced ethanol production from enzymatically treated steam‐exploded rice straw using extractive fermentation

Alcohol fermentation of an enzymatic hydrolyzate of exploded rice straw was studied experimentally. Rice straw was treated under variable conditions, such as steam pressure and steaming time. The exploded rice straw was separated into water‐soluble material, methanol‐soluble lignin, Klason lignin, and a mixture of cellulose and a low molecular weight substance. The effects of steam explosion on the characteristics of the exploded rice straw were clarified from the point of view of the amounts of extractive components. Steam explosion was found to be effective for the delignification of rice straw and for increasing its susceptibility to enzyme hydrolysis and alcohol fermentation. The polysaccharides (cellulose and hemicellulose) in the rice straw treated at a steam pressure of 3.5 MPa with a steaming time of 2 min were hydrolyzed almost completely into monosaccharides, (ie glucose and xylose) by a mixture of Trichoderma viride cellulase (Meicelase) and Aspergillus aculeatus cellulase (Acucelase). The enzymatic hydrolyzate of exploded rice straw was converted into ethanol efficiently by Pichia stipitis and the ethanol yield from sugar was about 86%(w/w) of the theoretical value. The ethanol concentration in a membrane bioreactor coupled with a pervaporation system reached 50 gdm^?3 and was about five times higher than that in the culture broth. The energy efficiency (ratio of combustion energy of ethanol produced to energy for steam explosion) reached a maximum value at a pressure of 3.5 MPa for 2 min. © 2001 Society of Chemical Industry     

纤维素燃料乙醇生产技术研究进展

纤维素燃料乙醇已成为下一代燃料乙醇的必然发展方向。文章综述了近年来以木质纤维素为原料生产燃料乙醇的关键技术,重点对物理法、化学法、蒸汽爆破法、生物法等木质纤维素原料预处理技术,酸水解、酶水解等水解(糖化)技术,以及直接发酵法、水解发酵两步法、同步水解发酵法等发酵工艺进行了总结,并指出了未来纤维素乙醇的产业化过程中必须解决的关键问题和发展趋势。     

Investigation of cellulose convertibility and ethanolic fermentation of sugarcane bagasse pretreated by wet oxidation and steam explosion

Sugarcane bagasse was pretreated by wet oxidation (WO) at 195 °C for 15 min under either alkaline, neutral or acidic conditions, and by steam explosion (STEX) at 205 °C for 10 min. Alkaline WO was more favourable than neutral and acidic WO for the following enzymatic hydrolysis of cellulose, giving 792 g kg^?1 glucose yield after 48 h. The enzymatic hydrolysis of the fibres in the whole slurry was inhibited by inhibitory compounds contained in the prehydrolysate in comparison with the hydrolysis of the washed solid fibres in buffer. The inhibition increased proportionally with formic acid concentration in the pretreated liquid fraction. Cellulose conversion was higher for simultaneous saccharification and fermentation (SSF) than for separate hydrolysis. The highest SSF conversion (829 g kg^?1) was obtained for the material treated by alkaline WO. The fermentability of the prehydrolysates by Saccharomyces cerevisiae was evaluated. Stronger inhibition of ethanolic fermentation was observed in the prehydrolysate obtained by steam explosion. The inhibition was more noticeable for the volumetric productivity than for the ethanol yield. The volumetric productivity was reduced by 94.5 and 91.2% for STEX and WO, respectively, whereas the ethanol yield was reduced only by 45.2 and 31.0%, correspondingly, for STEX and WO. Furan aldehydes seemed to be mainly responsible for the inhibition of the fermentation. Copyright © 2006 Society of Chemical Industry     

过氧甲酸预处理对甘蔗渣酶解和发酵的影响

以甘蔗渣(SCB)为原料, 经过氧甲酸(PAP)预处理后加入酶进行水解, 并以水解液发酵产乙醇, 考察预处理时过氧化氢(HPP)浓度变化对甘蔗渣酶解和乙醇得率的影响。实验结果表明: 在甘蔗渣PAP预处理过程中, HPP与甲酸(FAP)体积比为1∶1时, 预处理甘蔗渣(PAP-SCB-1)的木质素脱除率达84.30%;在纤维素酶用量为10 FPIU/g(以预处理后的甘蔗渣质量计)时, PAP-SCB-1水解72 h葡萄糖得率为98.71%, 较单独过氧化氢预处理甘蔗渣(HPP-SCB, 葡萄糖得率9.11%)和单独甲酸预处理甘蔗渣(FAP-SCB, 葡萄糖得率7.06%), 分别提高了9.84和12.98倍; PAP-SCB-1水解液经24 h发酵后, 乙醇得率为84.06%, 比HPP-SCB(76.20%)和FAP-SCB(75.15%)均有增加。对预处理前后物料的化学成分变化、比表面积和结晶度进行测定, 结果显示: 经PAP预处理后可以显著脱除甘蔗渣中的木质素, 木质素的量由未经预处理的21.27%降低到10%以下; 比表面积和结晶度都有提高, PAP-SCB-1的比表面积和结晶度分别为13.01 m²/g和54.18%, 是HPP-SCB的10.66和1.11倍, FAP-SCB的11.39和1.15倍。     

驯化树干毕赤酵母发酵产乙醇的研究

以玉米秸秆蒸汽爆破液为底物培养基,通过逐步提浓的方式对树干毕赤酵母菌株（Pichiastipitis）NLP23进行耐抑制物驯化,驯化后的菌株对汽爆液中甲酸和乙酸等抑制物的耐受能力可分别达到2．70g／L和3．54g／L,较出发菌株分别提高253．40％和277．80％。在含有57．34g/L木糖和13．84g／L葡萄糖的汽爆液中发酵42h,糖利用率和乙醇得率可达到97．89％和65．83％,乙醇质量浓度为21．56g/L,同时生成4．16g/l木糖醇。玉米秸秆蒸汽爆破液中含有多种抑制酵母生长和发酵的有毒物质,主要是甲酸、乙酸、乙酰丙酸、糠醛和羟甲基糠醛,其中甲酸和乙酸含量较高,是影响树干毕赤酵母NLP23发酵汽爆液的主要抑制物。     

Extraction of bioethanol from fermented sweet sorghum bagasse by batch distillation

Extraction of bioethanol, a potential alternative to fossil fuel in the transport industry, from sweet sorghum stems [Sorghum bicolor (L.) Moench] using solid-state fermentation (SSF) technology has become a popular research topic worldwide. Because SSF technology can directly convert fermentable sugars into target products without juice squeezing and water input, this method can potentially reduce energy and water consumption. However, ethanol extraction from fermented sweet sorghum bagasse requires further investigation. We used batch solid-state distillation to investigate the optimal operating parameters in a distillation column (diameter, 400 mm) via a single-factor experiment. Results showed that the optimal steam flow rate and loading height were 8-10 kg·h^?1 and 700-1,000 mm, respectively. Under optimal conditions, an energy consumption of 3.82 tons of steam per ton of ethanol and distillate concentration of 60.9% (v/v) were obtained. The pseudo-first-order rate equation was used to describe the distillation kinetics, and good correlations were obtained. Therefore, solid-state distillation can be effectively used to extract ethanol from fermented sweet sorghum bagasse.     

利用纤维素水解液中的纤维二糖发酵制备丁二酸

纤维素水解液中通常含有纤维二糖。本文考察了Actinobacillus succinogenes NJ113利用纤维二糖厌氧发酵生产丁二酸的能力,并利用蔗渣纤维素制备纤维二糖作为碳源用于厌氧发酵生产丁二酸。3 L发酵罐厌氧发酵结果显示：以35 g/L纤维二糖作为碳源发酵制备丁二酸,其产量为23.51 g/L,产率达到67.17%;用含有18 g/L纤维二糖和17 g/L其它糖类的蔗渣纤维素水解液作为碳源发酵制备丁二酸,丁二酸的产量和产率分别为20.00 g/L和64.73%。因此,Actinobacillus succinogenes NJ113具有较强的利用纤维二糖生产丁二酸的能力,而且利用废弃的纤维素制备纤维二糖作为碳源高效、经济地发酵制备丁二酸具有可行性。     

An approach to optimization of enzymatic hydrolysis from sugarcane bagasse based on organosolv pretreatment

BACKGROUND: The organosolv pretreatment followed by enzymatic hydrolysis of the pretreated material and subsequent fermentation of the hydrolysate produced, was the strategy used for ethanol production from sugarcane bagasse. The effect of different operational variables affecting the pretreatment (the catalyst type and its concentration, and the pretreatment time) and enzymatic hydrolysis stage (substrate concentration, cellulase loading, addition of xylanase and Tween 20, and the cellulase/β‐glucosidase ratio), were investigated. RESULTS: The best values of glucose concentration (28.8 g L^?1) and yield (25.1 g per 100 g dry matter) were obtained when the material was pretreated with 1.25% (w/w) H₂SO₄ for 60 min, and subsequently hydrolyzed using 10% (w/v) substrate concentration in a reaction medium supplemented with xylanase (300 UI g^?1) and Tween 20 (2.5% w/w). Fermentation of the broth obtained under these optimum conditions by Saccharomyces cerevisiae resulted in an ethanol yield of 92.8% based on the theoretical yield, after 24 h. CONCLUSION: Organosolv pretreatment of sugarcane bagasse under soft conditions, and subsequent enzymatic hydrolysis of the pretreated material with a cellulolytic system supplemented with xylanase and Tween 20, is a suitable procedure to obtain a glucose rich hydrolysate efficiently fermentable to ethanol by Sacharomyces cerevisiae yeasts. Copyright © 2010 Society of Chemical Industry