酒精-沼气双发酵耦联工艺中SO42-的控制

SO2-4是"酒精-沼气双发酵耦联工艺"稳定运行的重要抑制物.在耦联工艺中,以中温厌氧出水代替自来水配料进行酒精发酵.通过无预糖化工序的"同步糖化发酵"技术研究,将酒精发酵的初始pH提高到6.0,H2SO4的消耗降低了50%,使SO2-4的质量浓度维持在3g/L的沼气发酵安全范围内.在进行高浓度酒精发酵时,糖化酶的添加量为每克木薯添加140U,发酵54h,最终酒精体积分数达14.7%.糖化酶添加量与常规酒精发酵用量相比,每克木薯增加了糖化酶20 U.     

废糟液全循环对自絮凝酵母糖酵解途径关键酶、胁迫相关代谢物及胞内组分的影响

旨在研究废糟液直接全循环对絮凝酵母乙醇发酵、糖酵解关键酶以及细胞组成的影响。在一有效容积1.5 L的搅拌式生物反应器中,使用葡萄糖为220 g/L,添加8 g/L酵母粉和6 g/L蛋白胨的培养基,以0.04 h?1的稀释率进行自絮凝颗粒酵母乙醇连续发酵。每隔3天将收集到的发酵液集中精馏处理,得到的废糟液用于配制发酵培养基。装置运行近20 d,实验结果表明,随着废液循环批次的增加,系统乙醇和生物量浓度明显降低,糖酵解途径3个关键限速酶:己糖激酶、6-磷酸果糖激酶和丙酮酸激酶不同程度受到抑制。为了应对废糟液中高沸点副产物积累导致的环境胁迫,维持细胞正常代谢,甘油和菌体胞内蛋白生物合成加强,碳水化合物积累减弱。这些研究结果对进一步研究高沸点副产物积累对酵母细胞乙醇发酵影响的机理和菌种的代谢工程改造,具有重要意义。     

菊粉乙醇发酵条件的响应面优化

采用响应面分析法对菊粉乙醇发酵条件进行优化,首先采用Plackett-Burman设计对影响菊粉乙醇发酵的7个因素进行筛选,获得影响最大的3个因素菊粉浓度、装液量和发酵时间.再利用响应面分析法对这3个因素进行优化,确定最佳发酵条件为菊粉浓度160.0g/L、装液量130mL、发酵时间84h,采用优化后的发酵条件进行发酵,乙醇浓度达到7.5%,取得了较好的优化效果.     

自絮凝颗粒酵母乙醇连续发酵耦合酵母回用工艺的研究

模拟现有酒精发酵行业普遍采用的多级发酵罐串联系统,建立了一套由三级串联操作的搅拌式发酵罐和两个沉降罐组成的反应器系统,以脱胚脱皮玉米粉双酶法制备的糖化液为发酵底物,培养基初始还原糖浓度为220g/L,添加(NH4)2HPO41.5g/L和KH2PO42.5g/L,以0.057h-1的恒定稀释速率流加,将自沉降浓缩后的酵母乳先后经活化和不活化两种方式处理并循环至第一级发酵罐,系统在两种操作条件下分别达到了拟稳态。实验结果表明活化处理对改善发酵工艺技术指标方面发挥了显著的作用,发酵终点乙醇浓度达到101g/L,还原糖和残总糖分别在3.2和7.7g/L左右,发酵系统的设备生产强度指标为5.77g/(L.h),与无酵母回用的搅拌式反应器系统中自絮凝颗粒酵母乙醇发酵工艺相比,提高了70%。     

休哈塔假丝酵母HDYXHT-01利用木糖生产乙醇的发酵工艺优化

采用Plackett-Burman (PB) 方法和中心组合设计 (Ccentral composit design,CCD) 对休哈塔假丝酵母Candida shehataeHDYXHT-01利用木糖发酵生产乙醇的工艺进行优化。PB试验设计与分析结果表明：硫酸铵、磷酸二氢钾、酵母粉和接种量是影响木糖乙醇发酵的4个关键因素,以乙醇产量为响应目标,采用CCD和响应面分析法 (Response surface methodology,RSM),确定了木糖乙醇发酵的最佳工艺为：硫酸铵1.73 g/L、磷酸二氢钾3.56 g/L、酵母粉2.62 g/L和接种量5.66％,其他发酵条件为：木糖80 g/L,MgSO4·7H2O 0.1 g/L,pH 5.0,培养温度30 ℃,装液量100 mL/250 mL,摇床转速140 r/min,发酵时间48 h,在该条件下发酵液中乙醇产量可以达到26.18 g/L,比未优化前提高了1.15倍。     

响应面法优化耐高温酵母生产高浓度乙醇

利用耐高温酵母GXASY-10菌株对木薯粉同步糖化(SSF)法生产高浓度乙醇的发酵条件进行了优化。在单因素实验的基础上,首先应用Plackett-Burman试验设计筛选影响酒精高温高浓度发酵的重要参数,采用最陡爬坡实验逼近最大酒精生产区域后,利用Box-Behnken设计确定重要参数的最佳水平。筛选结果表明,影响酒精产量的重要参数是液化时间、糖化酶用量和初始木薯粉(底物)浓度。最佳工艺条件为:液化时间为35min,糖化酶添加量为1.21AGU/g底物,底物浓度为37.62%。20L发酵罐在此条件下(发酵温度37℃,转速100r/min)经过48h发酵,酒精浓度可达16.07%(V/W)。优化条件与初始条件相比较,酒精浓度提高了33%。     

多级串联悬浮床反应器系统中自絮凝颗粒酵母乙醇连续发酵耦合废糟液直接全循环使用的研究

在一套四级串联悬浮床生物反应器系统中,以双酶法制备的玉米粉糖化液为底物,进行了废糟液全循环条件下自絮凝颗粒酵母乙醇连续发酵的实验研究。在实验中,每隔5 d将从末级反应器收集到的发酵液集中精馏处理,得到的废糟液直接用于玉米粉调浆制糖。系统连续运行了120d,共进行了24批次实验,数据分析表明系统达到了平衡状态。在平均发酵时间为20h条件下,发酵终点乙醇浓度平均为11.7%（V/V）,残还原糖浓度平均为7.9g/L,装置运行平稳。这些工作为自絮凝颗粒酵母乙醇发酵耦合废糟液直接全循环使用、实现污染物源头减废、清洁生产奠定了理论和实验基础。     

泡盛曲霉CAU33固体发酵产β-1,3-1,4-葡聚糖酶发酵条件优化

利用单因素实验法优化了泡盛曲霉(Aspergillus awamori)CAU33利用农业废弃物固体发酵产β-1,3-1,4-葡聚糖酶的发酵条件。产酶的条件包括碳源种类、初始水分含量、氮源种类、初始p H、表面活性剂、培养温度和发酵时间。进一步运用响应面分析法优化了其中主要因素,得到最佳产酶条件为:啤酒糟为碳源、含水量为81.6%、吐温60添加量20g/L、大豆蛋白胨添加量25g/L、自然p H、35℃下培养6d。在优化后的发酵条件下,最大产酶水平达到40 832.9U/g。泡盛曲霉固体发酵产β-1,3-1,4-葡聚糖酶的酶活力高,工业化生产和应用潜力大。     

高浓度糖发酵制备乙醇

以树干毕赤酵母为发酵菌株,混合糖（木糖、葡萄糖）为发酵底物,通过培养基和培养条件的改变来确定树干毕赤酵母高糖浓度发酵时所需的条件。研究结果表明：在24h发酵周期内初始木糖质量浓度为63．0g／L较适宜;在36h发酵周期内初始木糖质量浓度为72．0g/L较适宜。24h发酵周期内,在36．0g／L木糖中添加的葡萄糖质量浓度以54．0g/L为最佳,发酵结束乙醇质量浓度达32．9g／L;36h发酵周期内,添加的葡萄糖质量浓度以72．0g／L为最佳,发酵结束乙醇质量浓度为36．9g／L。以（NH4）2SO4为N源时较适合戊糖发酵制备乙醇,（NH2）2SO4的最佳质量浓度为1．1g／L。发酵前8h摇床转速为90r／min,后16h为150r／min,乙醇质量浓度较高,可达17．5g/L。     

响应面法优化短短芽胞杆菌ch2-22的发酵工艺

在摇瓶发酵条件下,优化提高短短芽胞杆菌ch2-22芽胞浓度和抑菌活性的发酵培养基和培养条件。首先在单因素试验基础上进行响应面设计对ch2-22的发酵培养基进行优化,然后使用单因素试验方法确定最佳发酵条件,得到优化发酵培养基为淀粉35.05 g/L,豆饼粉26.08 g/L,蔗糖10 g/L,鱼粉5 g/L,Na Cl 1 g/L,Mg SO40.3 g/L,(NH4)2SO43 g/L,Mn SO40.1 g/L,K2HPO43 g/L,酵母膏1 g/L,Ca CO32 g/L。培养条件为温度32℃,转速180 r/min,装液量100 m L/500 m L,接种量4%,初始pH为7.0,发酵时间45 h。优化后的芽胞数量达到8.1×109cfu/m L,与优化前的芽胞数量(1.6×109cfu/m L)相比,提高了3.7倍,优化后发酵液效价达到3 350 IU/m L,提高了109%,高于同类菌株的2 000 IU/m L。     

利用功能基因组学方法研究酿酒酵母乙醇生物合成的调控机理

酿酒酵母在发酵生产乙醇的过程中存在的主要问题是前期高浓度底物葡萄糖的抑制和后期高浓度产物乙醇的抑制。功能基因组学技术的发展为从基因组水平上系统研究酿酒酵母乙醇生物合成的调控机理提供可能。本研究模拟工业发酵的条件,对酿酒酵母实验菌株BY4743为遗传背景的116个单基因缺失菌株进行了乙醇发酵试验,以发现基因和乙醇发酵的关系。结果表明乙醇对菌体得率系数高于平均值30%以上的基因缺失株有20株,其中高于50%以上基因缺失株有5株;低于平均值30%以上的基因缺失株有11株,其中低于45%以上的有5株。本研究为从整个酿酒酵母基因组水平上系统研究乙醇生物合成的调控机理建立了研究方法,提供了可行性验证。     

Fate of Fumonisin B(1) in Naturally Contaminated Corn during Ethanol Fermentation

Two lots of corn naturally contaminated with fumonisin B(1) (15 and 36 ppm) and a control lot (no fumonisin B(1) detected) were used as substrates for ethanol production in replicate 8.5-liter yeast fermentations. Ethanol yields were 8.8% for both the control and low-fumonisin corn, while the high-fumonisin corn contained less starch and produced 7.2% ethanol. Little degradation of fumonisin occurred during fermentation, and most was recovered in the distillers' grains, thin stillage, and distillers' solubles fractions. No toxin was detected in the distilled alcohol or centrifuge solids. Ethanol fermentation of fumonisin-contaminated corn coupled with effective detoxification of distillers' grains and aqueous stillage is suggested as a practical process strategy for salvaging contaminated corn.     

Fate of Fumonisin B1 in Naturally Contaminated Corn during Ethanol Fermentation

Two lots of corn naturally contaminated with fumonisin B₁ (15 and 36 ppm) and a control lot (no fumonisin B₁ detected) were used as substrates for ethanol production in replicate 8.5-liter yeast fermentations. Ethanol yields were 8.8% for both the control and low-fumonisin corn, while the high-fumonisin corn contained less starch and produced 7.2% ethanol. Little degradation of fumonisin occurred during fermentation, and most was recovered in the distillers' grains, thin stillage, and distillers' solubles fractions. No toxin was detected in the distilled alcohol or centrifuge solids. Ethanol fermentation of fumonisin-contaminated corn coupled with effective detoxification of distillers' grains and aqueous stillage is suggested as a practical process strategy for salvaging contaminated corn.     

Fermentation of biomass sugars to ethanol using native industrial yeast strains

In this paper, the feasibility of a technology for fermenting sugar mixtures representative of cellulosic biomass hydrolyzates with native industrial yeast strains is demonstrated. This paper explores the isomerization of xylose to xylulose using a bi-layered enzyme pellet system capable of sustaining a micro-environmental pH gradient. This ability allows for considerable flexibility in conducting the isomerization and fermentation steps. With this method, the isomerization and fermentation could be conducted sequentially, in fed-batch, or simultaneously to maximize utilization of both C5 and C6 sugars and ethanol yield. This system takes advantage of a pH-dependent complexation of xylulose with a supplemented additive to achieve up to 86% isomerization of xylose at fermentation conditions. Commercially-proven Saccharomyces cerevisiae strains from the corn-ethanol industry were used and shown to be very effective in implementation of the technology for ethanol production.     

An industrial level system with nonisothermal simultaneous solid state saccharification,fermentation and separation for ethanol production

To alleviate the problems of low substrate loading, nonisothermal, end-product inhibition of ethanol during the simultaneous saccharification and fermentation, a nonisothermal simultaneous solid state saccharification, fermentation, and separation (NSSSFS) process was investigated; one novel pilot scale nonisothermal simultaneous solid state enzymatic saccharification and fermentation coupled with CO₂ gas stripping loop system was invented and tested. The optimal pretreatment condition of steam-explosion was 1.5 MPa for 5 min in industrial level. In the NSSSFS, enzymatic saccharification and fermentation proceeded at around 50 °C and 37 °C, respectively, and were coupled together by the hydrolyzate loop; glucose from enzymatic saccharification was timely consumed by yeast, and the formed ethanol was separated online by CO₂ gas stripping coupled with adsorption of activated carbon; the solids substrate loading reached 25%; ethanol yields from 18.96% to 30.29% were obtained in fermentation depending on the materials tested. Based on the pilot level of 300 L fermenter, a novel industrial-level of 110 m³ solid state enzymatic saccharification, fermentation and ethanol separation plant had been successfully established and operated. The NSSSFS was a novel and feasible engineering solution to the inherent problems of simultaneous saccharification and fermentation, which would be used in large scale and in industrial production of ethanol.     

Effect of soy skim from soybean aqueous processing on the performance of corn ethanol fermentation

The feasibility of using soy skim, a co-product of the aqueous processing of soybeans, in ethanol production from corn was evaluated. Specific growth rates were compared when Saccharomyces cerevisiae was grown in soy skim and peptone-yeast extract media supplemented with glucose. Such soy skim was proved to be a good nitrogen source for yeast growth. Next, fermentation of dry-ground corn to ethanol using soy skim as the media was simulated on 1.5-L scale. Replacing water with soy skim increased the initial ethanol production rates by 4-32% while final ethanol yield was about 39 g/100 g dry corn, similar to the result when water was used. Solid and protein contents in the finished beer increased with the addition of soy skim. Thus, replacing water in corn-ethanol fermentation with soy skim is feasible, and may improve the economics of both aqueous soybean processing and corn ethanol fermentation.     

Production of ethanol by coupling fermentation and solvent extraction

Summary A new technology of fermentation is proposed. The inhibitor product is removed continuously by coupling fermentation and solvent extraction. Applied to ethanol fermentation this technology is suitable to any case where the terminal product is inhibitory.The proposed technology uses both plug flow reactor and liquid-liquid extraction to achieve continuously the extractive fermentation of ethanol. The solvent used for liquid-liquid extraction is dodecanol. A new reactor was used. It is a column packed with a porous material . The fermentation broth is pulsed (a) to increase the interfacial area between the liquid medium and the dodecanol, and (b) to: decrease the gas hold up.Alcoholic fermentations were performed on glucose syrup at 35°C using Saccharomyces cerevisiae, with adsorbed cells as reference, with adsorbed cells and extractive fermentation. The results show that the fermentation is substantially improved. By this new method the ethanol productivity was multiplied by 5 and a solution of 407 g/l of glucose was totally fermented with a yeast which cannot normally transform more than 200 g/l glucose.     

Grain pearling and very high gravity (VHG) fermentation technologies for fuel alcohol production from rye and triticale

A SATAKE laboratory abrasive mill was used for rye and triticale grain processing. About 12% of dry grain mass was removed after three and five successive abrasions for triticale and rye, respectively. Starch contents in the pearled grain were increased by 8·0% for triticale, and by 7·1% for rye. The pearled rye and triticale were ground and fermented by active dry yeast for fuel alcohol production by very high gravity (VHG) fermentation at 20°C. VHG technology was applied to increase final ethanol concentrations in the fermentors from 9·5–10·0% (v/v) (normal gravity) to 12·9–15·1% (v/v). The grain pearling process coupled with VHG technology further raised the ethanol concentration to 15·7–16·1% (v/v). Partial removal of outer grain solids in an alcohol plant would improve plant efficiency and decrease energy requirements for mash heating, mash cooling, and ethanol distillation.     

Ethanol production in a continuous fermentation/membrane pervaporation system

The productivity of ethanol fermentation processes, predominantly based on batch operation in the U.S. fuel ethanol industry, could be improved by adoption of continuous processing technology. In this study, a conventional yeast fermentation was coupled to a flat-plate membrane pervaporation unit to recover continuously an enriched ethanol stream from the fermentation broth. The process employed a concentrated dextrose feed stream controlled by the flow rate of permeate from the pervaporation unit via liquid-level control in the fermentor. The pervaporation module contained 0.1 m² commercially available polydimethylsiloxane membrane and consistently produced a permeate of 20%–23% (w/w) ethanol while maintaining a level of 4%–6% ethanol in a stirred-tank fermentor. The system exhibited excellent operational stability. During continuous operation with cell densities of 15–23 g/l, ethanol productivities of 4.9–7.8 gl^–1 h^–1 were achieved utilizing feed streams of 269–619 g/l glucose. Pervaporation flux and ethanol selectivities were 0.31–0.79 lm^–2 h^–1 and 1.8–6.5 respectively.