木质纤维素乙醇发酵研究中的关键点及解决方案

该文综述了近年来木质纤维素材料发酵生产乙醇的研究进展，提出了在木质纤维素原料发酵生产乙醇过程中的两大关键点，一是减少和消除原料预处理过程中抑制物及其有害影响；二是含木糖、葡萄糖、甘露糖、半乳糖等多种物质的混合物同时作为底物的乙醇发酵。在对已有研究结果比较和分析基础上，提出相应的解决方案。首先要采取综合的预处理过程，达到提高木质纤维素水解率的同时减少发酵抑制物；然后是提高发酵菌种对混合糖底物的利用能力和发酵生成乙醇的能力以及对抑制物的耐受性，以提高木质纤维素发酵生产乙醇的转化率。     

超低酸预处理结合酶解提高玉米秸秆糖化效率

为提高纤维素酶解糖化的效率,该文采用超低浓度硫酸水解预处理废弃玉米秸秆。重点考察了不同酸浓度、反应温度、反应时间条件下超低浓度酸水解及后续酶解的总还原糖、葡萄糖及木糖的产率,详细叙述了总还原糖及各种单糖在酸水解及酶解过程中的转化规律,通过正交试验确定酸水解的最佳工况为酸浓度0.1%,反应温度160℃,反应时间55 min,搅拌180 r/min,固液比1∶10。酸水解后进行酶解(酶用量5%,pH值4.6,时间24 h,温度50℃)得到还原糖、葡萄糖、木糖产率分别为56.22%、16.97%、18.83%。通过红外光谱和纤维素分析仪对酸水解和酶解后的残渣进行分析可知,纤维素、半纤维素的转化率分别为88.52%、95.18%,进一步计算还原糖、葡萄糖、木糖的转化率为88.11%、44.86%、72.49%。该方法较大程度避免了还原糖在酸水解过程中的降解,保证了半纤维素还原糖的转化效率,进一步提高了总还原糖的产率,为超低酸水解在燃料乙醇领域提供了新的应用途径。     

木质纤维素乙醇研究进展（英）

木质纤维原料制取乙醇在实际应用中具有较大的发展前景。该文主要综述了纤维素乙醇的预处理、水解和发酵三个过程。此外,比较了不同预处理方法的效率,还概括了当前纤维素乙醇的研究进展,包括发酵菌株、基因工程、发酵过程（SHF和CBP）和发酵过程与分离过程的整合。最后,还对纤维素乙醇在今后的发展提出一些建议。     

白酒丢糟的酸酶联合水解糖化工艺

为充分利用白酒丢糟资源,探讨了酸酶联合水解法对其进行糖化以获得可发酵糖的可行性。以木糖和还原糖浓度为指标,研究温度、固液比、混合酸浓度和时间等因素对酸解效果的影响;在此基础上分析纤维素酶对酸解残渣(AHR)的酶解历程,并利用扫描电镜(SEM)、红外光谱(FTIR)和X-衍射(XRD)技术考察不同水解阶段丢糟的结构特性变化。结果表明,丢糟在温度为100℃、固液比为1:12g/mL和酸浓度为2.0%的条件下经混合酸水解120min可获得59.32g/L还原糖和6.49g/L木糖,该酸解阶段的半纤维素和纤维素转化率分别为77.38%和62.50%,木质素溶出率为43.50%。AHR在纤维素酶用量为4000U/g原料、温度为45℃和pH值为4.8的条件下继续作用2.5h可获得13.27g/L还原糖,该酶解阶段的纤维素转化率为66.67%,酶解率高达90.73%。结构特性研究表明,水解作用前后的丢糟形貌结构变化明显,孔隙率和比表面积增加,有利于纤维素酶对AHR中纤维结晶区的作用。FTIR和XRD结果显示,水解前后的特征组分所对应的吸收峰强度发生了变化,相对结晶度逐渐提高。白酒丢糟经酸酶联合水解作用转化为可发酵糖具有可行性。该研究可为丢糟生物质发酵制备乙醇提供理论基础。     

纤维素超低酸水解产物的分析

基于木质纤维素类生物质水解为人类提供乙醇等能源、化工产品的重要性，该文以定量滤纸模拟生物质的主要组分－纤维素，以其超低酸水解试验得到的最佳工况下液体产物和固体残渣为研究对象，通过高效液相色谱(HPLC)结合KS-802糖柱对产物质中糖的种类进行了划分，以GC-MS对副产物进行定性，并通过扫描电镜(SEM)及热重和工业分析元素分析对固体残渣作了从表观到内部的研究，发现纤维素超低酸水解主要生成纤维四糖、三糖和二糖等低聚糖和葡萄糖及果糖，反应副产物有糠醛、羟甲基糠醛、乙酰丙酸及一些小分子酮、醛类和酸类极性化合物，水解残渣已完全改变了原始形貌，热裂解活性增强，残留物中碳和灰分含量有较大增加，最后根据分析结果探讨了纤维素超低酸水解的反应途径，为木质纤维素类生物质水解技术的规模化利用打下基础。     

Mn2+和卵磷脂对以植物秸秆为原料发酵生产乙醇的影响

考察了不同浓度Mn2+和卵磷脂对以玉米秸秆水解糖为原料发酵生产乙醇的影响。结果表明,在酵母发酵培养基中同时添加0.6 mmol/L Mn2+和60 mg/L卵磷脂进行乙醇发酵,终点乙醇浓度可达15.46%(V/V),乙醇的得率达到49%。     

玉米秸秆稀酸水解糖化法影响因子的研究

该文通过分析水解还原糖成分,研究了温度、时间、稀酸浓度、固形物含量对水解效率的影响;并结合残渣组分分析研究了稀酸水解过程中纤维素、半纤维素降解的规律.研究表明:在试验条件下玉米秸秆的水解主要以半纤维素为主,随着温度、稀酸浓度和水解时间的增加,纤维素的水解逐渐增强.当水解温度超过1000C后,水解得糖率迅速增加,超过110℃,纤维素开始水解;玉米秸秆的水解在前40 min就基本完成,过长的水解时间对水解效率的提高意义不大;水解的酸浓度应控制在1.5%左右;玉米秸秆在低固含物的条件下,水解效率相对较高.方差分析结果显示:温度、酸浓度对水解效率的影响最大,温度和固含物的影响次之,粒径的影响不显著;温度和酸浓度、温度和时间的交互作用对试验结果有影响显著.根据方差分析,最佳水解条件A3B2C3D1E1:温度125℃、时间80 min、稀酸浓度1.5%、固形物含量7.5%,在该条件下水解还原糖得率为32.71%.     

SO2蒸汽预处理对黑松制取生物乙醇的影响

以加拿大不列颠哥伦比亚省甲虫致死黑松为原料,通过SO2催化蒸汽爆破方式对其进行预处理,并对预处理后的原料进行酶水解和乙醇发酵研究,以考察预处理方式结合酶水解对黑松制取乙醇的影响。结果表明,较低的SO2吸收率和原料含水率（干基）影响了预处理效果;水洗对预处理后原料的酶水解效率没有显著的影响;当水解底物质量浓度由20 mg/mL增加到150 mg/mL时,水解率均在42%左右,底物质量浓度的增加纤维素的水解率没有受到显著影响。从发酵结果看,黑松经汽爆预处理后,不进行水洗处理更有利于后续发酵。分步糖化发酵乙醇得率可以达最大乙醇得率的66%,而同步糖化水乙醇得率为55%。由此可见,低浓度SO2催化汽爆预处理低湿度黑松不能得到较好的预处理效果,需进一步优化。     

CO一步法C. autoethanogenum发酵产乙醇的工艺研究

合成气/CO发酵制备燃料乙醇是一项具有吸引力的新技术,为促进C.autoethanogenum在该技术中的应用,对C.autoethanogenum的乙醇发酵工艺及过程参数进行了研究。结果表明,C.autoethanogenum代谢木糖的产物以乙酸为主,只产生少量乙醇;与无机氮源相比较,C.autoethanogenum在含有机氮源的培养基中生长迅速,菌体浓度高。在3 L发酵罐中进行C.autoethanogenum的批式发酵试验,采用木糖生长-CO发酵两步法,乙醇主要在CO发酵阶段产生,最高乙醇质量浓度为1.71 g/L;发酵罐经改进之后,采用CO一步法发酵,虽然得到的菌体浓度降低了,但是发酵时间延长,最高乙醇质量浓度达到7.36 g/L,而乙酸质量浓度在整个发酵过程中均低于1.1 g/L。此外,研究发现发酵液的pH值和氧化还原电位ORP与乙酸/乙醇产物分布密切相关,尤其是pH值。上述研究结果可为C.autoethanogenum发酵CO生产乙醇的中试放大提供参考。     

以菊芋粉为原料同步糖化发酵生产燃料乙醇

利用粟酒裂殖酵母（Schizosaccharomyces pombe）能发酵菊芋未水解糖液高产乙醇的特点提出了以菊芋粉为原料,同步糖化发酵生产燃料乙醇的新工艺。在摇瓶中考察了原料预处理方法、原料浓度和初始pH值对乙醇发酵的影响,进而在5 L发酵罐中考察了未调控pH值和恒定pH值与通气情况对乙醇发酵的影响。结果表明：该菌株最适pH值为4.0;100目筛分的菊芋粉发酵效果良好,115℃灭菌处理优于121℃,在此条件下,菊芋粉浓度200 g/L时,乙醇产量达到66.58 g/L,理论转化率为85.88%;发酵液pH值下降对乙醇发酵没有影响,通入适量氧气会导致乙醇产量的下降,这表明粟酒裂殖酵母进行乙醇发酵时不需要供氧;通入氮气保持厌氧环境不能显著提高乙醇产量,不通气进行乙醇发酵也达到高的转化率,因此在工业生产中,不必保持厌氧发酵环境。在此基础上,对菊芋粉补料发酵进行了试验,补料至菊芋粉终浓度为300 g/L,发酵终点乙醇浓度为94.81 g/L,理论转化率为81.54%。这些研究工作,为以菊芋为原料的燃料乙醇工业化生产提供技术依据。     

蒸汽爆破玉米芯水解液脱毒及其发酵生产燃料丁醇

为探究以玉米芯为原料生产燃料丁醇的最佳工艺技术,该研究对蒸汽爆破玉米芯水解液的脱毒方式及脱毒后的水解液的丙酮丁醇发酵进行了研究。结果表明:D301树脂对玉米芯水解液进行脱毒的综合效果最好,甲酸、乙酸和总酚的脱除率分别达到60%、46.04%和56.31%,香草醛脱除率为100%,对糠醛和5-HMF的脱除率分别达到了82.95%和87.52%;同时总糖的损失率为4.38%。D301树脂脱毒后的水解液经C.acetobutylicum CICC 8016发酵丁醇和总溶剂产量分别为5.2和7.5 g/L,葡萄糖和总糖的利用率分别达到100%和73.67%。当D301树脂脱毒的玉米芯水解液初始糖的质量浓度为50 g/L时,丁醇和总溶剂(丙酮、丁醇和乙醇)的质量浓度分别达到最大9.7和14.6 g/L。该研究为利用玉米芯工业化生产燃料丁醇提供了可靠的技术支持。     

酶法复合脱毒提高玉米秸秆水解液丁醇发酵效率

利用玉米秸秆发酵产丁醇在生物质转化领域具有明显优势。为解除玉米秸秆水解液中多种有毒物质对微生物生长的抑制及对发酵产量的影响,该研究摒除常用的理化脱毒法,选择高效环保的酶法脱毒以实现溶剂高产。研究结果表明:通过优化漆酶和甲酸脱氢酶添加量以去除水解液中酚类和甲酸,单独添加漆酶5 U/m L、甲酸脱氢酶1 U/m L,水解液发酵的丙酮-丁醇-乙醇(acetone-butanol-ethanol,ABE,总溶剂)产量分别为1.03和1.11 g/L。再在活性炭的辅助下形成高效酶法复合脱毒体系,经复合脱毒处理的水解液发酵后丁醇产量达2.90 g/L,总溶剂ABE产量达到4.4 g/L,比未作处理的对照组发酵产量高出约5倍,实现了生物质的高效转化。可为玉米秸秆水解液发酵生产燃料丁醇提供参考。     

玉米秸秆酶解上清液厌氧发酵产氢工艺优化

该文主要以粒度小于0.088 mm秸秆粉的酶解上清液为底物与热预处理后的活性污泥进行厌氧发酵产氢试验,以累积产氢量为考察指标,基于响应面Box-Behnken模型研究不同影响因素对玉米秸秆酶解上清液厌氧发酵产氢的影响,对玉米秸秆酶解上清液厌氧发酵产氢工艺进行优化。结果表明:温度、初始p H值和还原糖浓度三因素中,温度和还原糖浓度对玉米秸秆酶解上清液厌氧发酵产氢的影响最大。采用Box-Behnken模型获得的最佳产氢条件为:温度38.32℃,初始p H值4.93,还原糖浓度20.70 mg/m L,最大产氢量685.59 m L,此时最大产氢率为57.13 m L/g(玉米秸秆)。通过试验验证,实际最大产氢量为659.24 m L,产氢率为54.94 m L/g(玉米秸秆),与模型预测值相比,相对误差为3.84%,说明该模型具有较好的拟合性。该优化工艺可为后期连续流状态下的生物制氢系统提供参考。     

棉秆脱毒水解液发酵生产2,3-丁二醇的工艺优化

2,3-丁二醇是一种重要的化工产品,利用棉秆水解液替代淀粉原料制备2,3-丁二醇可保证粮食安全并降低成本。该文以棉秆稀酸水解液为基础,研究了其中糠醛和苯酚微波辅助加热-活性炭吸附的脱毒条件,优化结果为：活性炭用量1%、微波功率330 W、作用时间10 min。在此工艺条件下,糠醛的去除率为81.2%,苯酚的脱除率为92.3%,总糖的损失为10.6%。脱毒棉秆水解液为底物发酵生产2,3-丁二醇研究表明,水解液浓度为40 g/L时Klebsiella pneumoniae XJ-Li菌体浓度和2,3-丁二醇的产率最高,补料批式发酵可以缓解高浓度棉秆水解液对微生物生长与代谢的抑制作用。通过采用添加60 mg/L维生素C和维持发酵液pH值于5.5的复合调控方法,2,3-丁二醇的质量浓度达到了45.1 g/L,产率为0.45 g/g。发酵试验表明脱毒的棉杆水解液作为碳源发酵制备2,3-丁二醇具有可行性。     

Sucrose metabolism and exopolysaccharide production in wheat and rye sourdoughs by Lactobacillus sanfranciscensis.

The exopolysaccharide (EPS) produced from sucrose by Lactobacillus sanfranciscensis LTH2590 is predominantly composed of fructose. EPS production during sourdough fermentation has the potential to affect rheological properties of the dough as well as the volume, texture, and keepability of bread. Its in situ production by L. sanfranciscensis LTH2590 was demonstrated during sourdough fermentation after the hydrolysis of water soluble polysaccharides. In wheat and rye doughs with sucrose addition the concentration of fructose in the hydrolysate of polysaccharides was significantly higher than that in the hydrolysate of control doughs or doughs without sucrose addition. EPS production by L. sanfranciscensis in wheat doughs was confirmed by the determination of delta (13)C values of water soluble polysaccharides after the addition of naturally labeled sucrose, originating from C(3)- and C(4)-plants. In rye doughs, evidence for EPS production with the isotope technique could be demonstrated only by the determination of delta (13)C values of fructose from water soluble polysaccharides. In addition to EPS formation from sucrose, sucrose hydrolysis by L. sanfranciscensis in wheat and rye sourdoughs resulted in an increase of mannitol and acetate concentrations and in accumulation of glucose. It was furthermore observed that flour arabinoxylans were solublized during the fermentation.     

Effect of different forms of alkali treatment on specific fermentation inhibitors and on the fermentability of lignocellulose hydrolysates for production of fuel ethanol

Treatment with alkali, particularly overliming, has been widely used as a method for the detoxification of lignocellulose hydrolysates prior to ethanolic fermentation. However, the mechanisms behind the detoxification effect and the influence of the choice of cation have not been well understood. In this study, a dilute acid hydrolysate of spruce and an inhibitor cocktail consisting of six known inhibitors were used to investigate different alkali detoxification methods. The various treatments included the addition of calcium hydroxide, sodium hydroxide, potassium hydroxide, and ammonia to pH 10.0 and subsequent adjustment of the pH to 5.5 with either sulfuric or hydrochloric acid as well as treatment with the corresponding amounts of calcium, sodium, and potassium as sulfate or chloride salts at pH 5.5. An RP-HPLC method was developed for the separation of 18 different inhibitors in the hydrolysate, including furaldehydes and phenolics. Detection and quantification were carried out by means of UV, DAD, and ESI-MS in negative mode. Treatment of the spruce hydrolysate with alkali resulted in up to approximately 40% decrease in the concentration of furaldehydes. The effects on the aromatic compounds were complex. Furthermore, SFE was performed on the precipitate formed during alkali treatment to evaluate the inhibitor content of the precipitate, and the following RP-HPLC analysis implied that potential inhibitors were removed mainly through conversion rather than through filtration of precipitate. Parallel experiments in which sulfuric acid or hydrochloric acid was used for acidification to pH 5.5 after alkali treatment indicated that the choice of anion did not affect the removal of inhibitors. Detoxification with calcium hydroxide and ammonia resulted in better fermentability using Saccharomyces cerevisiae than detoxification with sodium hydroxide. The results from the experiments with the inhibitor cocktail indicated that the positive effects of alkali treatment are difficult to explain by removal of the inhibitors only and that possible stimulatory effects on the fermenting organism warrant further attention.     

Optimization of solid-state enzymatic hydrolysis of chestnut using mixtures of alpha-amylase and glucoamylase

Solid-state hydrolysis of starch present in chestnut was assayed in a single step with a mixture of a thermostable alpha-amylase and glucoamylase at three temperatures: 17 and 30 degrees C, for simultaneous hydrolysis and ethanol fermentation, and 70 degrees C, the optimal temperature for these enzymes. Total hydrolysis was only reached at the highest temperature, leading to a more concentrated hydrolysate than in submerged hydrolysis. Mass transfer limitations and starch retrogradation appear as the main causes for the incomplete hydrolysis of chestnut starch in solid-state operation at 17 and 30 degrees C. Even accepting that this limitation causes a 15% reduction of the yield of the hydrolysis with respect to the submerged process or the solid process at high temperature, solid-state hydrolysis at low temperatures seems to be adequate for simultaneous solid-state hydrolysis and fermentation processes.     

黄姜提取薯蓣皂甙元及葡萄糖的工艺研究

薯蓣皂甙元是黄姜中的主要活性成分，如何提高其得率和处理其水解滤液一直是实际生产中存在的问题。本文探讨了从黄姜水解液中同时得到薯蓣皂甙元和葡萄糖的提取工艺，首次提出了以中和作为主要脱盐方法从黄姜水解滤液中提取高纯度葡萄糖的工艺，确定了合适的中和、脱色和脱盐条件。结果表明，该工艺优于传统的薯蓣皂甙元提取方法，防止了水解滤液直接排放对环境造成的污染。     

Liquid chromatographic determination of protein acid hydrolysate content in blended soy sauce

Five methods were investigated for the determination of levulinic acid in soy sauce to determine the addition of protein hydrolysate, mainly acid hydrolysate of defatted soybeans. Best results were obtained by using liquid chromatography (LC) with 0.004 M HClO4 as the mobile phase and bromcresol purple as a post-column reagent. An innovative LC method with 0.1% H3PO4 as eluant was developed for determination of levulinic acid at 280 nm in soy sauce. This was the most time-saving method.     

废弃烟叶燃料酒精发酵工艺探索（简报）

以资源丰富、含糖量高的云南废弃烟叶为原料,通过正交试验,考察不同菌种、不同水平营养添加液和不同原料处理方法对能源酒精发酵的影响。试验结果表明：菌种和营养添加液水平对酒精发酵影响不明显,采用去除烟碱的预处理方法和半固态发酵方法,能显著提高燃料酒精产率;在此基础上,该文提出了较优的工艺流程,产物除燃料酒精外,还包括有重要用途的烟碱。