细胞固定化与硅橡胶膜渗透汽化分离耦合连续发酵制造乙醇

将硅橡胶膜(PDMS)渗透汽化分离与酵母细胞固定床耦合构成连续发酵系统,进行了乙醇发酵实验.发酵操作连续进行了378.5h,发酵液内的乙醇质量浓度最高达到了70g/L,启动PDMS膜分离后,降低并维持在50g/L左右;实验得到固定化颗粒和发酵液内的最高细胞浓度分别为1.76×1010个/g和9.8×108个/mL;乙醇体积产率3.67g/L·h,为游离连续发酵的2.1倍,葡萄糖消耗速率11.05g/L·h,为游离连续发酵的2.5倍;膜总通量400～690g/Cm2·h,乙醇通量为80～190g/m2·h,分离因子2.5～7.2,下游产品的平均质量浓度191.57g/L.     

细胞稀释强化硅橡胶膜生物反应器连续乙醇发酵

通过原位重力沉降分离酵母和渗透汽化分离乙醇构建了细胞稀释连续乙醇发酵的硅橡胶膜生物反应器。采用酿酒酵母,以0.05/h的细胞稀释率在膜生物反应器中实现了170 h的连续稳定乙醇发酵。重力沉降分离酵母对硅橡胶膜生物反应器产生的细胞稀释作用可以通过反应器内酵母自身生长得到平衡,发酵液细胞浓度稳定在5g/L。渗透汽化原位分离使发酵液内乙醇浓度维持在50 g/L。细胞稀释膜生物反应器连续发酵的乙醇体积产率达到1.63 g/(L.h),相对于同等工艺参数的细胞封闭循环膜生物反应器连续乙醇发酵细胞比产率提高了31%。     

葡萄糖对光滑球拟酵母发酵生产丙酮酸的影响

在30L发酵罐中研究了初始葡萄糖质量浓度和补料方式对光滑球拟酵母WSH-IP303发酵生产丙酮酸的影响．实验确定116.4g/L左右是较为适宜的初始葡萄糖质量浓度,发酵58h时丙酮酸质量浓度和产率分别为58.0g/L和0.516g/g．采用初始葡萄糖质量浓度为53.4g/L,发酵24h分批补料至葡萄糖总质量浓度为115g/L的培养方式,发酵64h时丙酮酸质量浓度和产率分别为60.2g/L和0.559g/g;采用初始葡萄糖质量浓度为62.6g/L,发酵24h开始连续补料至葡萄糖总质量浓度为115g/L的培养方式,发酵72h时丙酮酸质量浓度和产率分别为63.3g/L和0.586g/g,与葡萄糖总质量浓度相似(115g/L)的分批发酵相比,丙酮酸产量分别提高了3.8%和9.1%．实验结果表明适宜的初始葡萄糖质量浓度能促进光滑球拟酵母发酵生产丙酮酸;尽管葡萄糖补料培养可适度提高丙酮酸的产量及产率,但生产强度却有所下降．     

PDMS膜生物反应器中ABE发酵的实验研究

采用PDMS膜生物反应器和丙酮丁醇梭菌进行了丙酮-丁醇-乙醇(ABE)发酵实验研究.进行了2轮封闭循环连续发酵实验,发酵时间分别为264h和300h.2轮实验的发酵-膜分离耦合操作模式有区别:第1轮实验仅在白天(每天8:00～20:00)运行膜渗透汽化同步分离提取ABE产物(间断耦合);第2轮实验则在前192 h发酵阶段全程运行膜同步分离(连续耦合),之后采用和第1轮相同的间断耦合发酵模式.结果表明,第2轮发酵细胞的生长状况和发酵能力好于第1轮;2轮实验发酵液平均浓度分别为1.781g/L和1.884g/L;第2轮实验丁醇体积产率、得率系数、丁醇总产量分别比第1轮提高了95.2％、14.2％、121.2％.对发酵动力学行为进行了初步分析.     

补充营养物对硅橡胶膜生物反应器长期封闭循环发酵性能的影响

通过向发酵液中补充营养物质,研究了补充营养物对膜生物反应器长期封闭循环发酵行为的影响;实验得到的细胞浓度为7.10 g/L,细胞存活率为0.90,发酵液内的平均乙醇浓度为68.6 g/L,乙醇产率为1.85 g/(L·h),乙醇得率为0.411 g/g,乙醇比产率为0.286 g/(g·h),与不补充营养物质的实验相比,发酵性能有所提高;实验发现补充营养物质,没有提高细胞浓度和细胞存活率,而是增强了细胞的生命活性,提高了细胞产乙醇的能力。     

通气量对酿酒酵母GGSF16高浓度乙醇发酵的影响

在5 L发酵罐中,研究了以260 g/L葡萄糖为底物,不同通气量对酿酒酵母GGSF16高浓度乙醇发酵的影响。结果表明,与厌氧条件相比,通入适当的空气是高浓度乙醇发酵过程中重要的控制参数,能够缩短发酵时间,增加酵母细胞量,提高酵母细胞的存活率和乙醇耐受性。然而,乙醇产率与单位质量的酵母细胞消耗葡萄糖和生成乙醇的能力降低。最适通气量为80 m L/min,酵母细胞干重为14.16 g/L,发酵强度为3.93 g/(L·h),比厌氧分别提高104%和70.1%,终点乙醇浓度为117.9 g/L,乙醇产率为0.452 g/g(发酵效率87.8%)。     

乙醇对法夫酵母发酵合成虾青素的影响

利用摇瓶培养法夫酵母JMU-MVP14菌株,通过乙醇传感器检测乙醇含量,研究乙醇对法夫酵母细胞生长及虾青素合成的影响。结果表明,在10 g/L和20 g/L葡萄糖初始培养基中分别添加1～2 g/L乙醇时,均可提高虾青素的产量;而添加高浓度乙醇,则会抑制细胞生长和虾青素的合成。以10 g/L葡萄糖为初始培养基,控制发酵中恒定乙醇浓度为2 g/L,虾青素质量浓度可达到50.1 mg/L,比只添加2 g/L乙醇提高了20.3%,且虾青素细胞产率达到13.1 mg/g,葡萄糖的利用率能达到77.2%以上,说明恒定乙醇控制策略更有利于法夫酵母虾青素的合成。在10 g/L初始葡萄糖浓度,恒定乙醇浓度2 g/L,并每24 h补加5 g/L葡萄糖的优化条件下,虾青素产量最大值达到55.3 mg/L,虾青素细胞产率为19.7 mg/g,比对照组(14.1 mg/g)提高了39.7%。     

用乳清粉生产乙醇的研究

通过细胞固定化技术和混合发酵方式,对马克思克鲁维酵母和酿酒酵母混合发酵乳清粉产乙醇进行了研究。对比研究了游离细胞、混合游离细胞、单一固定化细胞、混合固定化细胞混菌和共固定化细胞混菌5种发酵方式发酵乳清粉生产乙醇。实验结果表明:混菌发酵要优于单一菌种发酵,固定化细胞发酵要好于游离细胞发酵。相对其他4种发酵方式,采用混合固定化马克思克鲁维酵母和固定化酿酒酵母混菌发酵方式,2种固定化菌种比例为1∶1时,利用乳清粉发酵可以获得最高的乙醇浓度5.3%vol,发酵周期最短为48h,乳糖利用率为1.64g(/L.h)。固定化细胞可以连续使用8个批次。     

棉纤维固定化酵母发酵产乙醇研究

为评估棉纤维固定化酵母生产乙醇的可行性,首先对棉纤维表面基团进行化学修饰,以提高载体对酵母细胞的吸附能力。其次分析了固定化发酵的稳定性,并从细胞数量角度定量分析固定化发酵和游离发酵的差异性。结果表明,经过F6-2试剂处理后的载体吸附效果有明显提升,其吸附比例达到83.1%,比未处理的固定化载体提高了15.9%。采用修饰的载体进行固定化发酵表现出较好的稳定性,发酵6批的平均转化率和产率分别为96.77%和5.22 g/(L·h),相比游离发酵,分别提高3%和51.7%。固定化发酵体系中,发酵6批后总酵母数为4.86×10⁸ CFU/mL,是游离发酵体系的2倍。结果表明,利用修饰的棉纤维进行固定化发酵具有明显的优势和应用可行性。     

固定化酵母细胞生物合成胞苷三磷酸工艺的优化

以胞苷一磷酸(CMP)为底物,采用响应面分析法对固定化酵母细胞发酵制备胞苷三磷酸(CTP)的影响因素进行优化,采用HPLC法对CTP定量。结果表明:在CTP生物合成中磷酸根离子和Mg2+浓度存在一个临界浓度低于临界浓度二者将限制CTP产率;在本实验范围内固定化酵母发酵制备CTP的最佳工艺为:20mL底物样液添加20g固定化酵母,CMP浓度100mmol/L、葡萄糖浓度100mmol/L、磷酸盐浓度250mmol/L、Mg2+浓度15mmol/L、pH6.1、反应时间2h,在此条件下CTP的产率可达67.89%。     

高浓度甘蔗汁酒精发酵过程的研究

研究高浓度甘蔗汁酒精发酵过程中糖的消耗,酒精生成以及酵母细胞生长的变化.采用液相色谱法测定各种糖的含量、气相色谱法测定酒精的生成量.试验结果表明,发酵12h时葡萄糖消耗速率最大(9.46(g/L)/h),14h时果糖、蔗糖、总糖消耗速率最大,分别为12.61(g/L)/h,12.30 (g/L)/h,20.59 (g/L)/h;发酵16h时酒精生成速率(7.89(g/L)/h)最大,细胞死亡率(22.9％)开始大于出芽率(12.6％),随后细胞活力逐渐下降.试验最终发酵酒精度12.8％vol,残糖22.50g/L,糖利用率90.9％,发酵效率79.69％.     

Characterization of membrane bioreactor for dry wine production

Characteristics of yeast growth and ethanol fermentation were examined in membrane bioreactor using a grape juice. After inoculation, batch fermentation was carried out for 24 h. When yeast growth reached the stationary phase, continuous fermentation was initiated. In continuous fermentation, a linear relationship was observed between cell concentration and dilution rate. In single-vessel membrane bioreactor, the cell concentrations of 18.7 g/l and 76.9 g/l (15 and 60 times higher than that of the batch fermentation, respectively) were observed at dilution rates of 0.1 h(-1) and 0.3 h(-1), respectively. The residual sugar concentration was higher than 10 g/l at the dilution rate of 0.1 h(-1), 0.2 h(-1) or 0.3 h(-1), therefore the single-vessel membrane bioreactor was not suitable for producing dry wine (sugar concentration: 4 g/l or less). In the double-vessel membrane bioreactor, it is most suitable to set the recycle ratio at 0.15 for keeping the sugar concentration below 4 g/l. The productivity of dry wine in the double-vessel membrane bioreactor was 28 times higher than that in the batch fermentation.     

桑叶对小曲质量的影响研究

研究了不同添加量的桑叶对小曲的微生物种类和数量、糖化力、液化力及发酵力的影响。结果表明,在一定范围内添加桑叶对小曲质量具有良好的促进作用,其合理添加量为9％,制成的小曲霉菌数达4．58×10^4个／g干曲,酵母数达4．77×10^3个／g干曲,细菌数达2．05×10^3个／g干曲。糖化力、液化力及发酵力分别达到498．2mg／g·h、0．82g／g·h、65．1％。     

Ethanol yield and volatile compound content in fermentation of agave must by Kluyveromyces marxianus UMPe-1 comparing with Saccharomyces cerevisiae baker's yeast used in tequila production

In tequila production, fermentation is an important step. Fermentation determines the ethanol productivity and organoleptic properties of the beverage. In this study, a yeast isolated from native residual agave must was identified as Kluyveromyces marxianus UMPe-1 by 26S rRNA sequencing. This yeast was compared with the baker's yeast Saccharomyces cerevisiae Pan1. Our findings demonstrate that the UMPe-1 yeast was able to support the sugar content of agave must and glucose up to 22% (w/v) and tolerated 10% (v/v) ethanol concentration in the medium with 50% cells survival. Pilot and industrial fermentation of agave must tests showed that the K. marxianus UMPe-1 yeast produced ethanol with yields of 94% and 96% with respect to fermentable sugar content (glucose and fructose, constituting 98%). The S. cerevisiae Pan1 baker's yeast, however, which is commonly used in some tequila factories, showed 76% and 70% yield. At the industrial level, UMPe-1 yeast shows a maximum velocity of fermentable sugar consumption of 2.27g·L(-1)·h(-1) and ethanol production of 1.38g·L(-1)·h(-1), providing 58.78g ethanol·L(-1) at 72h fermentation, which corresponds to 96% yield. In addition, the major and minor volatile compounds in the tequila beverage obtained from UMPe-1 yeast were increased. Importantly, 29 volatile compounds were identified, while the beverage obtained from Pan1-yeast contained fewer compounds and in lower concentrations. The results suggest that the K. marxianus UMPe-1 is a suitable yeast for agave must fermentation, showing high ethanol productivity and increased volatile compound content comparing with a S. cerevisiae baker's yeast used in tequila production.     

固定化酵母薯干原料酒精发酵中试研究

本文论述了固定化酵母薯干原料带渣酒精发酵的中试结果。中试生物反应器容积为１０００Ｌ，采用改性海藻酸钙凝胶为固定化酵母细胞的载体。中试研究中设计了较合理的中试工艺流程，优选出较佳的发酵条件，并考察了生物反应器的性能。试验获得了如下结果：中试装置已运转１９２天，固定化酵母仍保持高活力，并可继续运转；发酵成熟醪酒精含量达６．５～８．０％（ｖ），它比同期传统式酒精发酵高出０．３～０．４％；淀粉利用率达９１～９２％；固定化酵母生物反应器的乙醇生产能力为９．５ＫｇＥｔＯＨ／Ｍ￣３·ｈ，它可比传统式发酵罐的生产能力提高１０～１２倍。     

High gravity primary continuous beer fermentation using flocculent yeast biomass

The current work assessed a new immobilized cell reactor system throughout a long‐term (54 days) continuous primary fermentation of lager‐type wort of high specific gravity. The experiment was performed in a 4 L airlift bioreactor and immobilization of biomass was attained solely by flocculation. Despite the constant liquid agitation and washout of biomass, up to 53 g dry wt/L of yeast remained immobilized in the system. Two types of beer were produced without interrupting the reactor, based on two types of wort: a Pilsener type with high specific gravity of 15.6 ± 0.3°P; and a dark lager wort with specific gravity of 14.4 ± 0.03°P. Even during the inlet of high gravity wort, the desired attenuation was achieved without the need for either recirculation or an auxiliary second stage bioreactor. The specific saccharide consumption rate was kept around 7.9 ± 0.4 g/L/h and ethanol productivity oscillated at 3.36 ± 0.2 g/L/h for nearly a month. During this period the volumetric productivity of the current bioreactor reached 1.6 L beer/L/day. The green beers produced from the Pilsener and dark lager worts met the standards of regular finished primary beer fermentation. The productivity of diacetyl through the entire experiment could be correlated to the free amino nitrogen consumption rate. Copyright © 2014 The Institute of Brewing & Distilling     

Continuous Fermentation of Worcestershire Sauce by Trickle Bed Bioreactor Using Comb-Shaped Ceramic Carriers

A two-step fermentation process using Saccharomyces cerevisiae OC-2 (wine yeast) was studied. The first step, to multiply and immobilize yeast cells, was carried out by batch method (7 days) in a trickle bed bioreac-tor. In a second step, the substrate was continuously fermented by immobilized and free yeast cells in the same bioreactor. Continuous fermentation resulted in retention of 3% (W/V) of ethanol, 50 mg/L of isoamyl alcohol and 6 mg/L of β-phenethyl alcohol (the major aromatic components of fermented Worcestershire sauce). Maximum ethanol productivity was retained at 4.1–4.2 g/L/hr under continuous operation with immobilized yeasts, 3.2-fold higher than with the batch system.     

采用固定化酵母以带渣的薯干糖化醪进行酒精发酵的研究

研制了海藻酸钙膜片状固定化酵母填充床生物反应器,以带渣的低温蒸煮薯干糖化醪为基质,进行酒精发酵试验,获得了如下结果:淀粉利用率达90％;生物反应器的乙醇生产能力可达32mg乙醇/g凝胶·小时,以床层体积计为9.8g乙醇/l反应器·小时,为传统式酒精发酵罐乙醇生产能力的12倍;操作性能良好,反复活化运转三个月以上,发酵活力未见明显下降。