高浓度酒精连续发酵过程中振荡行为的模拟及填料弱化振荡的机理

在利用酿酒酵母进行高浓度酒精连续发酵中,观察到了一种持久稳定的振荡现象,表现为当严格控制发酵参数时,发酵罐中残糖浓度、酒精浓度和生物量浓度呈现大幅度的周期性波动。前期研究工作推测,细胞对环境胁迫的延迟反应是引起这种振荡行为的主要原因之一,并发现填料具有弱化这种振荡行为的作用,但描述这种振荡行为的动态模型尚未建立,填料弱化振荡的机理也尚不清楚。通过引入时滞加权函数,建立了预测高浓度发酵（VHGF）条件下振荡行为的过程动态模型。并在前期实验研究基础上,分别采用3种性质不同的填料,通过考察其弱化振荡行为的差异,在一定程度上揭示填料对振荡行为弱化作用的机理。实验结果表明：采用木块填料能明显地弱化振荡现象,金属丝网环和聚氨酯填料则不能弱化振荡现象;填料弱化振荡现象的原因与酵母细胞生长微环境的改变有关;利用聚氨酯作为填料能显著提高反应器中的生物量浓度,从而显著提高反应器系统的设备生产强度指标。     

气提法弱化超高浓度乙醇发酵过程中自絮凝酵母振荡行为

超高浓度(very high gravity,VHG)连续乙醇发酵过程中的振荡行为会导致发酵终点的乙醇浓度振荡和降低,是VHG连续乙醇发酵面临的主要问题。研究表明,游离酵母Saccharomyces cerevisiae 4126和自絮凝酵母BHL01在VHG连续乙醇发酵过程中,生物量、残糖、乙醇、甘油等发酵参数都呈现130～145 h的周期性振荡,且絮凝酵母发酵体系的平均乙醇浓度和乙醇生产强度都明显高于游离酵母体系。在絮凝酵母VHG连续乙醇发酵过程中利用发酵尾气气提分离乙醇,酵母细胞振荡行为被明显弱化,达到拟稳态状态,并且使发酵液中的残糖浓度控制在0.1 g·L^-1以下,平均乙醇浓度为110.87 g·L^-1,乙醇产率达到2.99 g·L^-1·h^-1。因此,本研究为弱化VHG连续乙醇发酵中的参数振荡行为提供了新的技术手段。     

高浓度乙醇连续发酵振荡过程中代谢通量分析及诱发机理

在利用普通酿酒酵母进行高浓度乙醇连续发酵的实验中,发现了一种长周期、高振幅的振荡现象。利用流式细胞仪测定了振荡周期不同时点的细胞周期分布,表明这种特殊的振荡现象和酵母细胞周期的同步化不相关。对一个完整振荡周期中不同时点的代谢通量分析,发现胞内碳通量在代谢网络中的分布也呈现出和胞外残糖、乙醇和生物量浓度类似的振荡过程。分析酵母细胞代谢活性与胞外发酵体系乙醇浓度的时程关系,表明酵母细胞对乙醇抑制的延迟反应是诱发这种振荡行为的主要因素,胞内海藻糖积累与乙醇生成同步,进一步支持了这一观点。     

CSTR中生化反应振荡行为研究

考虑在生化反应中细胞收率系数Ｙｘ／ｓ随物料浓度变化这一事实，改进了原有的发酵动力学方程，同时首次运用分岔理论研究了乙醇连续发酵过程中存在的振荡现象。结果表明：在低糖浓度条件下，振荡的周期和幅值以及振荡出现的入口浓度阈值都将随着稀释率和入口糖浓度的变化而变化     

振荡流态化现象及应用

研究了轻颗粒振荡流态化现象,通过对颗粒在振荡体中运动方程的求解,得到了单颗粒运动模型及重颗粒克服重力漂浮或者轻颗粒逆浮力下沉的临界参数关系式.用五种粒径、三种不同密度的轻颗粒实验数据及梁百申、邓勇有关重颗粒的数据验证了模型,结果表明实验值与理论值吻合较好.同时,从理论上得到流体在振荡情况下颗粒床层松散的数学表达式,并以实验进行了验证.振荡流态化反应器用于固定化细胞发酵酒精,可解决发酵过程产生的CO_2气体附着在酵母载体表面导致发酵过程操作条件恶化的问题,提高了操作稳定性和发酵反应速度.     

氧化还原电位控制下自絮凝酵母高浓度乙醇发酵

控制自絮凝酵母高浓度乙醇发酵过程的氧化还原电位（oxidoreduction potential, ORP）能降低环境胁迫对细胞的影响,提高乙醇生产强度和乙醇收率。实验考察了初始糖浓度为200、250、300 g·L^-1及ORP控制为-100、-150 mV和不控制的乙醇发酵情况。结果表明控制ORP的发酵过程,生物量和细胞存活率均高于不控制的系统,相应的发酵速度得到了提高,但是乙醇对糖的收率存在最优值。在实验设定初始糖浓度最高的300 g·L^-1的发酵过程中,控制ORP为-150 mV时,取得了最大的净乙醇生成量和乙醇对糖的收率。ORP控制改变了絮凝颗粒的粒径分布,运用多元线性拟合,发现ORP对絮凝的影响是正向的。ORP改变了发酵液中生物量及代谢物的浓度而间接影响了细胞的絮凝状况。     

固定化酵母乙醇萃取发酵研究

以十二烷醇为萃取剂，对固定化酵母乙醇萃取发酵进行了研究。探讨了萃取剂对酵母细胞的毒性，以及萃取剂用量、搅拌转速、基质浓度等因素与乙醇萃取发酵的关系，并测定了萃取过程中乙醇的分配系数。为固定化酵母乙醇发酵与溶剂萃取耦合新工艺的开发研究提供了资料。     

环流反应器中固定化酵母乙醇连续发酵研究

制备了一种重化固定化酵母颗粒，可有效防止发酵过程中固定化颗粒上浮，用于气升式内环流反应器，可与器内流体形成良好循环。同时在气升式内环流反应器内对固定化酵母乙醇连续发酵过程进行了研究，得到发酵最优控制条件。通过与其它发酵器对比发现，内环流反应器用于固定化酵母乙醇发酵过程，可大大提高乙醇产率。     

乙醇发酵与渗透汽化在硅橡胶膜生物反应器中的耦合强化

用硅橡胶膜生物反应器(SMBR)实验研究了发酵-渗透汽化的耦合性能。发酵微生物采用酿酒活性干酵母,所用的碳源为工业级葡萄糖。间歇发酵过程由于产物抑制作用在乙醇浓度达到90g稬-1时就趋于停滞,而经耦合渗透汽化膜分离后,发酵罐内的乙醇浓度迅速降低并维持在40g稬-1,且发酵在此浓度下可以连续稳定地进行。 在SMBR运行达到稳态后,乙醇的体积产率为1.5gL-1h-1。SMBR中所用的聚二甲基硅氧烷(PDMS)复合膜由实验室自行制备,它能稳定分离含有酵母细胞的发酵液。当发酵液中乙醇浓度为92.7~49.5g稬-1时,PDMS复合膜的总通量为1490~1164g穖-2h-1,分离因子为6.9~7.8,与分离相同进料浓度的清洁模型溶液相比分别平均高出31%和14%。乙醇发酵和渗透汽化在硅橡胶膜生物反应器中能够相互耦合并得到强化。     

膜生物反应器封闭循环乙醇发酵残液的处理技术研究

硅橡胶膜生物反应器封闭循环乙醇发酵系统能长时间平稳运行，发酵罐中乙醇浓度、酵母浓度和乙醇产率维持稳定，乙醇产率较传统间歇发酵可提高4．5倍，但系统长期运行的周期受制于酵母细胞的老化。我们目前的实验连续运行记录是500h。系统运行周期结束时，发酵罐中的残液成为需要处理的发酵废弃物，其总量约为传统间歇发酵工艺的1／6，但性态差别很大。对残液进行后处理，有效利用废弃物并实现发酵工艺的废掖零排放，成为该系统的又一主要工艺技术问题。本文对系统运行及工艺技术、残液性态、残液处理工艺路线和设备进行了研究，提出了解决方案。     

Continuous alcohol production from whey permeate using immobilised cell reactor systems

The present paper reports the performance of a bioreactor packed with alginate-entrapped Kluyveromyces marxianus NCYC179 for continuous fermentation of whey permeate to ethanol. A maximum ethanol productivity as 28.21 gl^?1 h^?1 was attained at D=0.42h^?1 and 75% lactose consumption (substrate feed rate in the inflowing medium was 200 g lactose I^?1). However, the higher dilution rates (0.6-1.Oh^?1) resulted in poor productivities and higher substrate washout in the effluent samples. The maximum specific ethanol production (qpi) and maximum specific lactose uptake (qsi) of the immobilised Kluyveromyces marxianus NCYC179 was found to be 3.88g ethanol/g immobilised cell/hx10^?2 and 8.75g lactose consumed/g immobilised cell/hx10^?2 respectively. A bead size of 2.5 mm in diameter and activation period of 24h of alginate beads in lactose solution (10%) prior to their packing in column reactor were found to support the efficient working of the bioreactor. The immobilised cell bioreactor system was operated continuously at a constant dilution rate of 0.15h^?1 and 10% lactose for 562 h without any significant change in the efficiency (varied from 84 to 88% of theoretical) and viability of the entrapped yeast cells (dropped from 84 to 81%).     

Performance of different immobilized‐cell systems to efficiently produce ethanol from whey: fluidized batch,packed‐bed and fluidized continuous bioreactors

BACKGROUND: The bioconversion of whey into ethanol by immobilized Kluyveromyces marxianus in packed‐bed and fluidized bioreactors is described. Both batch and continuous cultures were analyzed using three different strains of K. marxianus and the effect of the operating mode, temperature, and dilution rates (D) were investigated. RESULTS: All immobilized strains of K. marxianus (CBS 6556, CCT 4086, and CCT 2653) produced similar high yields of ethanol (0.44 ± 0.01 g EtOH g^?1 sugar). Significant variations of conversion efficiencies (66.1 to 83.3%) and ethanol productivities (0.78 to 0.96 g L^?1 h^?1) were observed in the experiments with strain K. marxianus CBS 6556 at different temperatures. High yields of ethanol were obtained in fluidized and packed‐bed bioreactors continuous cultures at different D (0.1 to 0.3 h^?1), with the highest productivity (3.5 g L^?1 h^?1) observed for D = 0.3 h^?1 in the fluidized bioreactor (87% of the maximal theoretical conversion), whereas the highest ethanol concentration in the streaming effluent (28 g L^?1) was obtained for D = 0.1 h^?1. Electronic micrographs of the gel beads showed efficient cell immobilization. CONCLUSION: Batch and continuous cultivations of immobilized K. marxianus in fluidized and packed‐bed bioreactors enable high yields and productivities of ethanol from whey. Copyright © 2012 Society of Chemical Industry     

乙醇连续发酵与膜超滤耦合过程研究

采用ＣＡ－Ｔｉ复合管式膜组件与发酵耦合操作，用啤酒酵母从葡萄糖连续发酵乙醇是一个从不稳态到稳态的过程。其稳态的最佳操作参数为：初始葡萄糖浓度１４０ｇ／Ｌ，稀释率０．３ｈ~（－１），轴出比０．６３８，相应的细胞浓度２×１０~９ｃｅｌｌ／ｍＬ，葡萄糖利用率９２％，生产率２２ｇ／（Ｌ·ｈ）。当细胞浓度增加到一定程度时，乙醇生产率和酵母比生长速率的增长减缓。反应器中细胞的浓度由稀释率和轴出比决定。     

Near-wall porosity characteristics of fixed beds packed with wood chips

Packed beds of fuel wood chips are commonly found in thermal conversion processes such as combustion or gasification. Wood chips in particular are mostly used as fuel for small-scale domestic heating boilers but also for commercial-scale combustion units. The characterization of spatial voidage distribution inside the wood chip beds is of great importance for flow and reactor modelling. This study focuses on the radial porosity variations of cylindrical beds of three different types of commercially available wood chips including chips classified as G30 size class. The conventional technique of consolidating packed beds with a resin was chosen as the experimental procedure. The radial voidage distribution in different cylindrical beds is determined by image analysis of sections of the solidified packings. Additionally, a packing of monosized spheres was investigated in order to assess the selected procedure in comparison with widely available literature data for spheres. The results are discussed and summarized in a mathematical expression correlating the radial voidage distribution depending on average wood chip size, packing core porosity and dimensionless distance from the tube wall.     

Hydrolysis of milk lactose in a packed bed reactor system using immobilized yeast cells

BACKGROUND: Hydrolysis of lactose with β‐D‐galactosidase is one of the most promising biotechnological applications in the food industry because of its use in the production of low lactose milk products and whey hydrolysis. To overcome the problem of enzyme extraction from cells due to the intracellular nature of β‐D‐galactosidase and the poor permeability of the cell membrane to lactose, permeabilization of yeast cells was investigated. Permeabilized whole cells have been claimed to have an advantage over more pure enzyme preparations. In view of the advantages of immobilized cell systems over free cell systems, permeabilized cells were immobilized by an entrapment method in calcium alginate gel. A packed bed reactor together with this immobilized cell system has been used for hydrolysis of milk lactose in a continuous system. RESULTS: Different process parameters (temperature, substrate feed rate, biomass load and time‐course) were optimized to maximize lactose hydrolysis. The immobilized yeast cells (300 mg dry wt) resulted in 87.2% hydrolysis of milk lactose at 30 °C and flow rate 7 mL h^?1 in a packed bed reactor system. CONCLUSION: This convenient and relatively inexpensive method of immobilization, resulting in high hydrolysis potential in a continuous system, indicates that permeabilized yeast cells have the potential for the production of low lactose milk and milk products. Copyright © 2010 Society of Chemical Industry     

Elimination of ethanol inhibition of yeast growth by a multistream ethanol feed in a multistage tower fermenter

The effect of several types of ethanol feed into a multistage tower fermenter, under conditions of oxygen supply rate limitation, on growth and physiological activity of the yeast Candida utilis was studied. The measurements were made at steady states of continuous culture for single values of dilution rate, temperature and pH in all stages of the fermenter and with a constant total sum of ethanol supplied. It was found that the type of ethanol feed significantly affects both the economic parameters associated with biomass production and the cell physiology in the individual stages of the fermenter.     

同时进行发酵和分离的CO_2气提、活性炭吸附乙醇发酵动力学研究——(三)固定化细胞发酵

研究了应用固定化酵母同时进行发酵和分离的 CO_2气提的乙醇间歇发酵和连续发酵。从实验结果发现,无论是间歇还是连续操作,无论是改变稀释率还是改变进口底物浓度,在实验范围内,发酵速率都基本上保持常数,即,固定化酵母发酵时,发酵液中少量游离酵母的影响可以忽略,固定化酵母密度在发酵过程中不会发生很大变化。由此而导出的简化模型可以满意地关联稳态操作时的实验数据,并进而推算动态响应曲线。     

Continuous ethanol production from wood hydrolysate by chemostat and total cell retention culture

Chemostat and total cell retention cultures with internal filter system ofSaecharomyc.es cerevisiae H1-7 were carried out to produce ethanol from wood hydrolysate. Maximum ethanol productivity obtained in a chemostat with the aeration rate of 1 vvm was 3.79 g/(L·h). This was 20% higher than that in a chemostat without aeration. However, the substrate was not completely consumed at the dilution rate with the maximum productivity. The realistic productivity, which has higher than 99% conversion rate of substrate, was. 2.95 g/(L·h). The maximum productivity in the total cell retention culture was 6.65 g/(L·h) at the dilution rate of 0.19 h¹ and the residual glucose concentration was negligible.