The fermentation of mixtures of D‐glucose and D‐xylose by Candida shehatae,Pichia stipitis or Pachysolen tannophilus to produce ethanol

The fermentation of mixtures of D ‐glucose and D ‐xylose by three non‐traditional yeasts: Candida shehatae (ATCC 34887), Pachysolen tannophilus (ATCC 32691) and Pichia stipitis (ATCC 58376) have been studied to determine the optimal strain and initial culture conditions for the efficient production of ethanol. The comparison was made on the basis of maximum specific growth rate (µ_m), biomass productivity, the specific rates of total substrate consumption (q_s) and ethanol production (q_E) and the overall yields of ethanol and xylitol. All the experiments were performed in stirred‐tank batch reactors at a temperature of 30 °C. The initial pH of the culture medium was 4.5. The highest values of µ_m (above 0.5 h^?1) were obtained with P stipitis in cultures containing high concentrations of D ‐xylose. All three yeasts consumed the two monosaccharides in sequence, beginning with D ‐glucose. The values of q_s diminished during the course of each experiment with all of the yeasts. The highest values of the specific rates of total substrate consumption and ethanol production were obtained with C shehatae (for t = 10 h, q_s and q_E were above 5 g g^?1 h^?1 and 2 g g^?1 h^?1, respectively), although the highest overall ethanol yields were fairly similar with all three yeasts, at around 0.4 g g^?1. © 2002 Society of Chemical Industry     

Kinetic study on ethanol production using Saccharomyces cerevisiae ITV‐01 yeast isolated from sugar cane molasses

BACKGROUND: Bio‐ethanol production from renewable sources, such as sugar cane, makes it a biofuel that is both renewable and environmentally friendly. One of the strategies to reduce production costs and to make ethanol fuel economically competitive with fossil fuels could be the use of wild yeast with osmotolerance, ethanol resistance and low nutritional requirements. The aim of this work was to investigate the kinetics of ethanol fermentation using Saccharomyces cerevisiae ITV‐01 yeast strain in a batch system at different glucose and ethanol concentrations, pH values and temperature in order to determine the optimum fermentation conditions. RESULTS: This strain showed osmotolerance (its specific growth rate (µ_max) remained unchanged at glucose concentrations between 100 and 200 g L^?1) as well as ethanol resistance (it was able to grow at 10% v/v ethanol). Activation energy (Ea) and Q₁₀ values calculated at temperatures between 27 and 39 °C, pH 3.5, was 15.6 kcal mol^?1 (with a pre‐exponential factor of 3.8 × 10¹² h^?1 (R² = 0.94)) and 3.93 respectively, indicating that this system is biologically limited. CONCLUSIONS: The optimal conditions for ethanol production were pH 3.5, 30 °C and initial glucose concentration 150 g L^?1. In this case, a maximum ethanol concentration of 58.4 g L^?1, ethanol productivity of 1.8 g L^?1 h^?1 and ethanol yield of 0.41 g g^?1 were obtained. Copyright © 2010 Society of Chemical Industry     

Kinetics of Bioethanol Production Employing Mono‐ and Co‐Cultures of Saccharomyces cerevisiae and Pichia stipitis

The kinetics of bioethanol production using mono‐ and co‐cultures of Saccharomyces cerevisiae and Pichia stipitis with glucose, xylose, and glucose‐xylose sugar mixtures were investigated. A MATLAB® program was formulated for simulation of experimental results in order to get predicted values of ethanol production and sugar consumption and for kinetic parameter estimation. Kinetic parameters implied less extent of substrate and/or product inhibition when the co‐culture scheme of immobilized S. cerevisiae and free P. stipitis was employed for fermentation of mixed sugars. In addition, a high ethanol yield was achieved by applying this co‐culture strategy to wheat straw hydrolysates.     

Investigating glucose and xylose metabolism in Saccharomyces cerevisiae and Scheffersomyces stipitis via 13C metabolic flux analysis

¹³C metabolic flux analysis (¹³C? MFA) has been extensively applied in studying the glucose metabolism of yeast strains such as Saccharomyces cerevisiae and Scheffersomyces stipitis. Here, we tried to augment the previous fluxomic studies by applying ¹³C? MFA to rigorously investigate the metabolic flux distributions in S. stipitis and the recombinant S. cerevisiae strains when xylose was utilized as the sole carbon substrate. It was found that less carbon fluxes were diverted into the TCA cycle in S. stipitis than the recombinant S. cerevisiae strains. Compared to single sugar utilization, the co‐utilization of glucose and xylose by S. stipitis led to increased metabolic fluxes into the futile pathway and the TCA cycle, but did not improve sugar‐based ethanol yield. In addition, heterologous expression of xylose pathway in engineered S. cerevisiae strains may affect the glucose utilization. © 2013 American Institute of Chemical Engineers AIChE J, 59: 3195–3202, 2013     

Characterization of Saccharomyces cerevisiae immobilized onto chrysotile for ethanol production

Saccharomyces cerevisiae (CCT 3174 and commercial baker's yeast) was immobilized by adsorption onto chrysotile. The adsorbed yeast cells were easily washed out, but cells grown in situ were strongly attached by entrapment by chrysotile microfibres. In fermentation experiments with 30% (w/v) glucose solution, the immobilized cells showed a 1·3-fold increase in initial reaction velocity. For immobilized CCT 3174, the final ethanol yield was 26% higher than that with free cells. © 1998 Society of Chemical Industry     

Potential use of wine yeasts immobilized on Penicillium chrysogenum for ethanol production

BACKGROUND: Six different wine yeast strains (G1, X4, X5, P29, QA23, Uvaferm BC) were co‐immobilized in a natural, spontaneous way with Penicillium chrysogenum under special conditions without the need for an external support or chemical binder and provided six different ‘yeast biocapsules’. The purpose was to characterize and evaluate the biocapsules obtained in terms of yeast cell viability, ethanol production and reusability to assess their suitability for ethanol production and the development of industrially competitive alternative wine and beer production methods. RESULTS: Biocapsule size was found to decrease and quantity to increase with increasing shaking rate during the immobilization process. The fermentations were realized in YPD medium containing 18% (w/v) glucose with repeated fermentations reaching 10% (v/v) ethanol. X4 and Uvaferm BC biocapsules afforded at least seven uses with no significant decrease in ethanol production; P29 and QA23 biocapsules five times; and G1 and X5 three times each. Seemingly, ethanol production was directly related to the viability of yeast cells in the immobilizate under defined assay conditions. CONCLUSIONS: X4 and Uvaferm BC may be the most suitable yeast strains for autoimmobilization on P. chrysogenum with a view to their use in alcoholic fermentation processes. Copyright © 2011 Society of Chemical Industry     

高密度发酵在酒精生产中的应用进展

酒精不仅是重要的可再生能源,在食品、化工、医药等领域应用也极其广泛。高密度发酵（HCDF）能有效提高酒精产率降低能耗及废液排放,极具应用潜力。本文针对目前淀粉质原料HCDF中酵母选育、培养条件优化、工艺过程优化同步耦合及动力学模型的多元统计控制,综合分析了神经交叉网络-模糊控制和基于经验的实验系统等国内外最新进展,旨在为HCDF酒精生产工艺的进一步研究提供参考依据。     

Improvement of ethanol production in Saccharomyces cerevisiae by hetero‐expression of GAPN and FPS1 deletion

BACKGROUND: During anaerobic bioethanol fermentation of Saccharomyces cerevisiae, the main byproduct glycerol is essential to regulate redox balance (reoxidize NADH to NAD⁺), which is necessary to maintain cell growth and fermentation. Hetero‐expression of a NADP⁺‐dependent glyceraldehydes‐3‐phosphate dehydrogenase (GAPN) [EC.1.2.1.9] in S. cerevisiae could redirect the carbon flux from glycerol to ethanol involving a net oxidation of NADH. The present study investigates whether combination of GAPN hetero‐expression and glycerol exporter Fps1p disruption would result in less glycerol and more ethanol production without affecting growth rate during anaerobic fermentations. RESULTS: The results of anaerobic fermentations showed that the fps1Δ mutant with GAPN (named 4FG) produced 21.47% less glycerol and 9.18% more ethanol compared with a parental strain with a control plasmid, while the rates of growth and fermentation were not changed. Moreover, the engineered strain 4FG yielded less glycerol and acetic acid, and more ethanol than the control, fps1Δ mutant or with GAPN only. CONCLUSIONS: During anaerobic fermentations, hetero‐expression of GAPN restored the reduced grow rate of the fps1Δ mutant, and led to less byproducts and more ethanol production. This combination strategy could be used to modulate glycerol metabolism and optimize the anaerobic fermentation of S. cerevisiae. Copyright © 2011 Society of Chemical Industry     

纤维素乙醇生产重组酿酒酵母菌株的构建与优化研究进展

寻找化石能源的替代品以及开发和利用生物能源已引起国内外研究者的广泛关注。提高酿酒酵母利用来源广泛、贮存丰富的农林废弃物等木质纤维素原料生产燃料乙醇的效率是生物能源的重要研究内容,但是,重组酿酒酵母木糖发酵性能低是限制纤维素乙醇经济性的关键问题。本文总结了酿酒酵母中木糖代谢途径的构建和优化以及木糖转运对木糖利用的影响,分析了重组酵母利用纤维素水解液进行乙醇发酵的研究现状,并对进一步提高重组酿酒酵母纤维素乙醇生产效率的研究趋势进行了展望。目前国内外已经构建了可有效利用木糖产乙醇的重组酵母,但对其木糖代谢机制的研究还尚未深入,限制了重组菌株的定向改造。此外,目前缺少在纤维素生物质水解液发酵实际应用过程中对重组菌株的评价。因此,加强重组酵母菌株对木糖利用相关代谢调控机理的分析,注重多种抑制物对菌株发酵性能的影响,结合真实底物纤维素乙醇发酵过程进行重组菌株的构建和优化,从而进一步提高纤维素乙醇生产的经济性,是未来菌株构建的重要研究方向。     

整合型酿酒酵母菌株强化发酵和特性比较

为了提高木糖异构酶基因在重组酿酒酵母体内的稳定性,并比较不同真菌来源的木糖异构酶在木糖或者葡萄糖-木糖培养基的发酵利用特性,分别构建来自Piromyces sp.E2和Orpinomyces sp.的木糖异构酶基因的整合表达载体,利用同源重组将其整合进入呼吸缺陷型菌株的18S rDNA非转录区,结果测得Orpinomyces的木糖异构酶酶活活力为0.72 U/mg,比Piromyces的木糖异构酶酶活高2.8倍。在木糖培养基中发酵获得乙醇的得率分别为0.40 g/g和0.48 g/g。且整合入Orpinomyces的木糖异构酶基因菌株能获得最高酶活和乙醇得率。     

Co‐culture of Penicillium chrysogenum and Saccharomyces cerevisiae leading to the immobilization of yeast

BACKGROUND: Under appropriate conditions, Saccharomyces cerevisiae and Penicillium chrysogenum were found to co‐immobilize spontaneously with no need for external support or chemical binder. The main aims were to examine the interaction between yeast cells and fungal hyphae by electron microscopy and the death of the filamentous fungus because of direct contact between both microorganisms. RESULTS: Immobilization was accomplished by orbitally shaking at 28 °C a culture medium consisting of yeast nitrogen base buffered at pH 7 and containing gluconic acid as an available carbon source for the filamentous fungus not readily used by the yeast. The yeast biocapsules thus obtained were hollow, smooth, elastic, strong, creamy‐coloured spheres of variable size depending on the particular shaking rate and time of residence in the formation medium. Biocapsule walls consisted of yeast cells bound to fungal hyphae. Placing the biocapsules in fermentation medium caused yeast cells to colonize and invade all hyphae, thereby causing the fungus to die and remain as a mere support for yeast. CONCLUSIONS: The death of the fungus was probably effected by the yeast via a cell‐hypha contact‐mediated mechanism as shown by dialysis experiments. The yeast biocapsules can be reused with no loss of integrity or activity. The proposed immobilization method provides a simple, convenient, inexpensive alternative which affords yeast reuse. Copyright © 2011 Society of Chemical Industry     

Effects of light wavelength and intensity on the production of ethanol by Saccharomyces cerevisiae in batch cultures

BACKGROUND: Photoreceptors have been identified in Saccharomyces cerevisae, however, the influence of light on the performance of ethanol fermentation of S. cerevisiae is not yet clear. The aims of this study are to elucidate the influence of light wavelength and intensity on the growth and ethanol production of S. cerevisiae and to describe a novel two‐stage LED light process to optimize ethanol fermentation. RESULTS: Experimental results indicated that maximum biomass concentration X_max of the batch under red LED light increased monotonically with light intensity, and the optimal specific product yield Y_p/x was 13.2 g g^?1 at 600 lux. Maximum ethanol concentration P_max of the batch under blue LED light increased monotonically with light intensity, and the optimal Y_p/x was 18.4 g g^?1 at 900 lux. A novel two‐stage LED light process achieved maximum P_max, of 98.7 g dm^?3 resulting in 36% improvement compared with that of the batch in the dark. CONCLUSION: The light wavelength and its intensity significantly affected cell growth and ethanol formation of S. cerevisiae. Red LED light (630 nm) stimulated cell growth but slightly inhibited ethanol formation. In contrast, blue LED light (470 nm) significantly inhibited cell growth but stimulated ethanol formation. A novel two‐stage LED light process has been successfully demonstrated to optimize ethanol fermentation of S. cerevisiae. Copyright © 2009 Society of Chemical Industry     

Productivity enhancement of S‐adenosylmethionine in Saccharomyces cerevisiae using n‐hexadecane as oxygen vector

BACKGROUND: Saccharomyces cerevisiae is one of the main microorganisms that can produce S‐adenosylmethionine (SAM) from L‐methionine and ATP with high productivity. To satisfy the ATP requirement for SAM synthesis, sufficient oxygen should be supplied to the medium to improve aerobic metabolism in S. cerevisiae. In this study, n‐hexadecane used as oxygen vector for enhancement of SAM production by this yeast was investigated. RESULTS: N‐hexadecane was most favorable for cell growth and SAM synthesis in S. cerevisiae when added at the time of inoculation. It could increase glucose consumption, reduce ethanol accumulation, and ultimately improve biomass and SAM productivity in a fermentation process. In a bioreactor, the highest yield of SAM (2.27 g L^?1) was achieved in the presence of 4% (v/v) n‐hexadecane after 24 h of inoculation, which was 23.37% higher than the control (1.84 g L^?1). CONCLUSION: The addition of n‐hexadecane to cultures of S. cerevisiae significantly enhanced SAM production without increasing energy consumption, and has the potential for use in large‐scale fermentation processes to increase oxygen supply. Copyright © 2012 Society of Chemical Industry     

Conversions of olive mill wastewater‐based media by Saccharomyces cerevisiae through sterile and non‐sterile bioprocesses

BACKGROUND: Olive mill wastewaters (OMWs) are an important residue and several methods have been proposed for their treatment. RESULTS: Remarkable decolorization (~63%) and phenol removal (~34% w/w) from OMW was achieved. In glucose‐based flask sterile cultures, enrichment with OMWs increased ethanol and biomass production compared with cultures without OMWs added. Flask sterile and un‐sterilized cultures demonstrated similar kinetic results. Batch‐bioreactor trials performed showed higher ethanol and lower biomass quantities compared with the respective shake‐flask experiments, while cultures used under un‐sterilized conditions revealed equivalent results to the sterile ones. In non‐sterile bioreactor cultures, OMWs addition enhanced biomass production in comparison with culture with no OMWs added, whereas ethanol biosynthesis was not affected. The maximum ethanol quantity achieved was 52 g L^?1 (conversion yield per sugar consumed of 0.46 g g^?1) in a batch bioreactor non‐sterilized trial with OMW–glucose enriched medium used as substrate, that presented initial reducing sugars concentration at ~115 g L^?1. Fatty acid analysis of cellular lipids demonstrated that in OMW‐based media, cellular lipids containing increased concentrations of oleic and linoleic acid were produced in comparison with cultures with no OMWs added. CONCLUSIONS: S. cerevisiae simultaneously produced bio‐ethanol and biomass and detoxified OMWs, under non‐sterile conditions. © 2012 Society of Chemical Industry     

Effect of different fermentation parameters on bioethanol production from corn meal hydrolyzates by free and immobilized cells of Saccharomyces cerevisiae var. ellipsoideus

BACKGROUND: Bioethanol produced from renewable biomass, such as corn meal, is a biofuel that is both renewable and environmentally friendly. Significant scientific and technological investments will be needed to achieve substitution of conventional fossil fuels with alternative fuels. The ethanol fermentation of enzymatically obtained corn meal hydrolyzates by free and immobilized cells of Saccharomyces cerevisiae var. ellipsoideus yeast in a batch system was studied. The initial glucose and inoculum concentration and the time required for the efficient ethanol production were optimized taking into account parameters such as ethanol concentration, ethanol yield, percentage of the theoretical yield of ethanol and volumetric productivity in both immobilized and free cell systems. RESULTS: The yeast cells were immobilized in Ca–alginate by an electrostatic droplet generation method. An optimal initial inoculum concentration of 2% (v/v) and optimal fermentation time of 38 h for both immobilized and free yeasts were determined. An optimal initial glucose concentration of 150 g L^?1 for free system was achieved. At the initial glucose concentration of 176 g L no substrate or product inhibition were achieved with immobilized yeast. CONCLUSION: By immobilization of the yeast into Ca–alginate using the method of electrostatic droplet generation a superior system was realized, which exhibited lower substrate inhibition and higher tolerance to ethanol. The cells of S. cerevisiae var. ellipsoideus yeast entrapped in Ca–alginate showed good physical and chemical stability, and no substrate and product diffusion restrictions were noticed. Copyright © 2008 Society of Chemical Industry