Continuous Ethanol Fermentation at Very High Gravity in the Presence of <i>Saccharomyces cerevisiae</i>: A Bifurcation Analysis

In this paper, a study of the bifurcation analysis of fermentation of sugar to ethanol in presence of Saccharomyces cerivisiae at very high gravity is described. The bifurcation analysis was done for a concentration 280 gl&minus;1 of sugar, and the dilution rate was taken as the parameter of bifurcation. Two Hopf bifurcations (HB) at 280 gl&minus;1 were found. At dilution rate of 0.027 h&minus;1 the system exhibits damped oscillations and not sustained oscillations as previously reported because the system is close to a point of attraction, and we can attenuate these oscillations by the choice of initial conditions. The system exhibits sustained oscillations between the two Hopf Bifurcations, the first at 0.08028 h&minus;1 and the second at 0.04395 h&minus;1. These oscillations are the consequence of synchrony between the daughter and the mother yeast. Indeed, it is better to take a dilution rate between the two Hopf bifurcations (self sustained oscillations), in order to increase the ethanol productivity.

Improve Ethanol Yield Through Minimizing Glycerol Yield in Ethanol Fermentation of Saccharomyces cerevisiae

在 Saccharomyces cerevisiae 的乙醇发酵(S。cerevisiae ) ，甘油是主要副产品之一。这调查的目的是通过由使用在编码调停的隧道蛋白质的 FPS1，编码 glycerol-3-phosphate 脱氢酶之一的甘油出口和 GPD2 用一步舞基因代替是敲门外面的异种最小化甘油产量增加乙醇产量。分别地，编码 glutamate synthase 和夫酸安合成酶的 GLT1 和 GLN1 是用在 fps1 螖 gpd2 螖异种的二拍子的圆舞基因代替的 overexpressed。ZAL69 的发酵性质(fps1 螖:: LEU2 gpd2 螖:: URA3 ) 并且 ZAL808 (fps1 螖:: LEU2 gpd2 螖:: URA3 PPGK1-GLT1 PPGK1-GLN1 ) 在 microaerobic 下面，条件被调查并且与那些相比野类型(DC124 ) 。葡萄糖，乙醇的收益，甘油的收益，醋酸，和焦葡萄酸酸的消费被监视。与野类型相比，分别地，而甘油产量由 37.4 % 和 41.7 % 减少了， ZAL69 和 ZAL808 的乙醇收益被 13.17% 和 6.66 % 改进。同时，醋酸收益和焦葡萄酸酸产量与野类型相比戏剧性地减少了。我们的结果显示 S 的那 FPS1 和 GPD2 删除。cerevisiae 导致了减少的甘油产量并且在 fps1 螖 gpd2 螖增加了 GLT1 和 GLN1 的乙醇产量，而是同时的 overexpression 异种没比 fps1 螖 gpd2 螖有更高的乙醇收益变异。

Elementary Flux Mode Analysis for Optimized Ethanol Yield in Anaerobic Fermentation of Glucose with Saccharomyces cerevisiae

基本流动模式(EFM ) 分析基于构造细胞的网络在 Saccharomyces cerevisiae 在中央碳新陈代谢的新陈代谢的分析被使用。从新陈代谢的模型，计算生产乙醇的小径号码 37 远在 78 个基本流动模式之中把底层变换成乙醇。在里面基于这分析预言的新陈代谢的显型与设计紧张， KAM3 和 KAM11 的发酵表演适合很好的 silico，它证实了那 EFM 分析是有效的指导 Saccharomyces cerevisiae 的构造设计紧张，增加乙醇收益。

A novel aeration strategy in repeated-batch fermentation for efficient ethanol production from sweet sorghum juice

To improve the efficiency of ethanol production in a batch fermentation from sweet sorghum juice under a very high gravity(VHG)condition(~290 g/L of total sugar)by Saccharomyces cerevisiae NP01,repeatedbatch fermentation under an aerated condition(2.5 vvm for the first 4 h during every cycle)was done in a5-L fermenter.The average ethanol concentration(P),productivity(Qp)and yield(Yp/s)for five successive cycles were 112.31 g/L,1.55 g/L·h^-1 and 0.44,respectively with 80.97%sugar consumption.To complete sugar consumption,the total sugar of the juice was reduced to a high gravity(HG)level(~240 g/L).The results showed that yeast extract was not necessary for ethanol production,and aeration during every other cycle i.e.,alternating cycles,was sufficient to promote both yeast growth and ethanol production.The average P,Qpand Yp/svalues for eight successive cycles with aeration during alternating cycles were97.58 g/L,1.98 g/Láh and 0.41,respectively with 91.21%sugar consumption.The total fatty acids in the yeast cells under the aerated condition were^50%higher than without aeration,irrespective the initial sugar concentration,whereas the ergosterol contents under aeration condition were^29%to 49%higher than those without aeration.

High Ethanol Production by Marine-Derived Yeasts-<i>Saccharomyces cerevisiae</i>under Stress Pressures

For practical applications of bioethanol, the uses of both highly concentrated biomass materials and their effective fermentation by yeasts are indispensable in order to produce ethanol at low costs. However, as the saccharified products of those biomass generally contain abundant sugars, the yeasts are affected by the compounds and are inclined to decrease their physiological activities. In the process of fermentation, ethanol is gradually produced by the yeasts in the culture;the concentrated metabolic product also damages itself, and inhibition of the fermentation frequently occurs. The application of yeasts with high fermentative activities under stress pressures such as sugars and ethanol is thus desired for bioethanol production. In this study, various types of high-fermentative yeasts under stress pressures were isolated mainly from coastal waters in Japan and characterized. All yeast strains with high fermentative activities under 20% v/v ethanol were found to be Saccharomyces cerevisiae. The HK21 strain isolated from Tokyo Bay and identified as S. cerevisiae had the highest fermentation activity under 30% w/v sorbitol and under 20% v/v ethanol, and it produced approx. 70 g/l (9% v/v) ethanol from the 15% w/v glucose solution at 25 oC within 5 days.

Effect of magnetic field on the fermentation kinetics of <i>Saccharomyces cerevisiae</i>

Published literature has shown conflicting results regarding the effects of magnetic fields on the fermentation kinetics or cellular growth of various Saccharomyces cerevisiae strains. Here, two sets of experiments were conducted to characterize the role of magnetic fields on cell growth and ethanol production during fermentation. The first experiment was completed for 25 h at a 2% dextrose loading rate under influence of homogeneous and non-homogeneous static magnetic fields on the order of 100 and 200 mT, respectively. The second experiment was completed for 30 h at a 6% dextrose loading rate under the influence of a non-homogeneous static magnetic field on the order of 200 mT. It was found that homogeneous magnetic fields have no significant effect on the yeast cell growth, while non-homogeneous static magnetic fields produced an increase (~ 8% over the control) in peak ethanol concentration with 2% dextrose loading.

Feeding a Saccharomyces cerevisiae fermentation product improves udder health and immune response to a Streptococcus uberis mastitis challenge in mid-lactation dairy cows

Background:We aimed to characterize the protective effects and the molecular mechanisms of action of a Saccharomyces cerevisiae fermentation product(NTK)in response to a mastitis challenge.Eighteen mid-lactation multiparous Holstein cows(n=9/group)were fed the control diet(CON)or CON supplemented with 19 g/d NTK for 45 d(phase 1,P1)and then infected in the right rear quarter with 2500 CFU of Streptococcus uberis(phase 2,P2).After 36-h,mammary gland and liver biopsies were collected and antibiotic treatment started until the end of P2(9 d post challenge).Cows were then followed until day 75(phase 3,P3).Milk yield(MY)and dry matter intake(DMI)were recorded daily.Milk samples for somatic cell score were collected,and rectal and udder temperature,heart and respiration rate were recorded during the challenge period(P2)together with blood samples for metabolite and immune function analyses.Data were analyzed by phase using the PROC MIXED procedure in SAS.Biopsies were used for transcriptomic analysis via RNA-sequencing,followed by pathway analysis.Results:DMI and MY were not affected by diet in P1,but an interaction with time was recorded in P2 indicating a better recovery from the challenge in NTK compared with CON.NTK reduced rectal temperature,somatic cell score,and temperature of the infected quarter during the challenge.Transcriptome data supported these findings,as NTK supplementation upregulated mammary genes related to immune cell antibacterial function(e.g.,CATHL4,NOS2),epithelial tissue protection(e.g.IL17C),and anti-inflammatory activity(e.g.,ATF3,BAG3,IER3,G-CSF,GRO1,ZFAND2A).Pathway analysis indicated upregulation of tumor necrosis factorα,heat shock protein response,and p21 related pathways in the response to mastitis in NTK cows.Other pathways for detoxification and cytoprotection functions along with the tight junction pathway were also upregulated in NTK-fed cows.Conclusions:Overall,results highlighted molecular networks involved in the protective effect of NTK prophylactic supplementation on udder health during a subclinical mastitic event.

Effects of Saccharomyces cerevisiae fermentation products on performance and rumen fermentation and microbiota in dairy cows fed a diet containing low quality forage

Background： A possible option to meet the increased demand of forage for dairy industry is to use the agricultural byproducts, such as corn stover. However, nutritional value of crop residues is low and we have been seeking technologies to improve the value. A feeding trial was performed to evaluate the effects of four levels of Saccharomyces cerevisiae fermentation product（SCFP; Original XP; Diamond V） on lactation performance and rumen fermentation in mid-lactation Holstein dairy cows fed a diet containing low-quality forage. Eighty dairy cows were randomly assigned into one of four treatments： basal diet supplemented with 0, 60, 120, or 180 g/d of SCFP per head mixed with 180, 120, 60, or 0 g of corn meal, respectively. The experiment lasted for 10 wks, with the first 2 weeks for adaptation.Results： Dry matter intake was found to be similar（P 〉 0.05） among the treatments. There was an increasing trend in milk production（linear, P ≤ 0.10） with the increasing level of SCFP supplementation, with no effects on contents of milk components（P 〉 0.05）. Supplementation of SCFP linearly increased（P 〈 0.05） the N conversion, without affecting rumen pH and ammonia-N（P 〉 0.05）. Increasing level of SCFP linearly increased（P 〈 0.05） concentrations of ruminal total volatile fatty acids, acetate, propionate, and butyrate, with no difference in molar proportion of individual acids（P 〉 0.05）. The population of fungi and certain cel ulolytic bacteria（Ruminococcus albus, R. flavefaciens and Fibrobacter succinogenes）increased linearly（P 〈 0.05） but those of lactate-utilizing（Selenomonas ruminantium and Megasphaera elsdeni） and lactate-producing bacteria（Streptococcus bovis） decreased linearly（P ≤ 0.01） with increasing level of SCFP. The urinary purine derivatives increased linearly（P 〈 0.05） in response to SCFP supplementation, indicating that SCFP supplementation may benefit for microbial protein synthesis in the rumen.Conclusions： The SCFP supplementation was effective in maintaining milk persistency of mid-lactation cows receiving diets containing low-quality forage. The beneficial effect of SCFP could be attributed to improved rumen function; 1）microbial population shift toward greater rumen fermentation efficiency indicated by higher rumen fungi and cel ulolytic bacteria and lower lactate producing bacteria, and 2） rumen microbial fermentation toward greater supply of energy and protein indicated by greater ruminal VFA concentration and increased N conversion. Effects of SCFP were dose-depended and greater effects being observed with higher levels of supplementation and the effect was more noticeable during the high THI environment.

Genetic control of protein and glucose-anabolic-enzyme syntheses by <i>Saccharomyces cerevisiae</i>in the fermentation of a Nigerian rice, <i>Oryza sativa</i>variety “Igbimo”

This work aimed at the control of the production of protein and glucose-anabolic-enzyme (GAE) by Saccharomyces cerevisiae during the fermentation of a Nigerian rice, Oryza sativa variety “Igbimo”. The yeast was mutated with ethylmethyl sulphonate. The variants and the parental yeast were separately inoculated into cooked rice and allowed to ferment at 27℃ for 7 days after which protein content and glucose-anabolic-enzyme synthesis were determined using Dinitrosalicylic acid and Biuret reagents techniques. Mutants with varying capacities to form protein and GAE were obtained. Glucose-Anabolic-Enzyme (GAE) activity ranged from 0.25 to 12.06 Units forming five groups (classes 1, 2, 3, 4 and 5) with the average activity of 0.52, 1.52, 2.28, 4.04 and 10.63 Units respectively compared with that (0.38 Unit) of the parent strain. All the mutants synthesized protein although many of them produced it at lower level while others at higher level than the wild-type. The highest (5.92 mg/mL) and lowest (0.10 mg/mL) levels protein producers are mutants 44 and 14 respectively. Mutants No. 4, 7, 22 and 78 formed total protein similar in concentrations (1.46, 1.46, 1.45 and 1.45 mg/mL) to that of the wild yeast (1.46 mg/mL). These three sets of protein concentration have ratios 4.1 (highest), 0.07 (lowest) and 1.0 (normal) to that of the parental yeast.

Protein Enrichment of Potato Peels Using Saccharomyces cerevisiae via Solid-State Fermentation Process

In order to add value to potato peels and also curb their environmental pollution problems, this study investigated the protein enrichment of potato peels with Saccharomyces cerevisiae via Solid-State Fermentation (SSF). SSF is a fermentation process which involves solid matrix and is carried out in absence or near absence of free water. SSF of potato peel mashed was carried out with S. cerevisiae at 30°C, pH of 5.5, moisture adjustment between 40 and 90%, addition of ammonium sulphate and urea salts as nitrogen supplements for the microorganisms for 3 days. The results showed that the percentage crude protein content of all the fermented samples increased significantly when compared with the unfermented sample. 40% moisture content adjustment and ammonium sulphate as nitrogen source gave the best result. The crude protein increased from 12.5% to 21.86%, which is 74.88% increment for ammonium sulphate supplementation, and 12.5% to 18.42%, which is 47% increment for urea supplementation. Therefore, the fermented peels could serve as good source of cheap protein enriched feed for livestock.

Asymmetric Synthesis of (-)-1-Trimethylsilyl-ethanol with Immobilized Saccharomyces Cerevisiae Cells in Water/Organic Solvent Diphasic System

Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied,The effects of shake speed,hydrophobictiy of organic solvent ,volume ratio of water phase to organic phase,pH value of aqueous phase and reaction temperature on the initial reaction rate,maximum yield and enantiomeric excess(ee) of the product were systematically explored,All the above-mentioned factors had significant infuence on the reaction.n-Hexane was found to be the best organic solvent for the reaction.The optimum shake speed,volume ratio of water phase to organic phase,pH value and reaction temperature were 150 r.min^-1,1/2,8 and 30℃ respectively,under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%,which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase ,To our best knowledge,this is the most satisfactory result obtained.

Influence of furfural concentration on growth and ethanol yield of Saccharomyces kluyveri

Furfural is an important inhibitor in ethanol fermentation process using lignocellulosic hydrolysates as raw materials. In order to find out the furfural concentration range in which furfural inhibits the fermentation process, we used one strain Saccharomyces kluyveri selected from soil and cultured in several different furfural content media under low glucose concentration condition. Experiment results showed that microorganism growth was stimulated and dry cell weight decreased when furfural concentration in the medium was 0.25 mg/ml. Furfural had negative effect on cell growth when its concentration was above 1.00 mg/ml. At the same time, the strain growed better and had a higher glucose consumption rate in 5% original glucose concentration condition than in 3% original glucose concentration condition. The results showed that appropriate exaltation of original glucose concentration in stalk hydrolysates will increase the strain resistance to furfural.

Fermentation of xylose to produce ethanol by recombinant Saccharomyces cerevisiae strain containing XYLA and XKS1

Fermentation of the pentose sugar xylose to produce ethanol using lignocellulosic biomass would make bioethanol production economically more competitive. Sac- charomyce cerevisise, an efficient ethanol producer, cannot utilize xylose because it lacks the ability to convert xylose to its isomer xylulose. In this study, XYLA gene encoding xylose isomerase (XI) from Thermoanaerobacter tengcongensis MB4T and XKS1 gene encoding xylulokinase (XK) from Pichia stipitis were cloned and functionally coexpressed in Saccharomyces cerevisiae EF-326 to construct a recombinant xylose-utilizing strain. The resulting strain S. cerevisiae EF 1014 not only grew on xylose as sole carbon source, but also produced ethanol under anaerobic conditions. Fermentations performed with different xylose concentrations at different temperatures demonstrated that the highest ethanol produc- tivity was 0.11 g/g xylose when xylose concentration was pro- vided at 50 g/L. Under this condition, 28.4% of xylose was consumed and 1.54 g/L xylitol was formed. An increasing fermentation temperature from 30℃ to 37℃ did not im- prove ethanol yield.

Influences of Lactiplantibacillus plantarum and Saccharomyces cerevisiae fermentation on the nutritional components,flavor property and lipid-lowering effect of highland barley

Highland barley is a well-known cereal in Qinghai-Tibet Plateau area with high nutritional value,which has been reported to be a health-promoting grain for the obesity and the diabetes.Fermentation by certain microbiota can improve the flavor property and nutritional characteristics.In the present study,Lactiplantibacillus plantarum and Saccharomyces cerevisiae were singly or jointly applied to ferment highland barley,and the profile of volatile substances and lipid-lowering effects of the respective extracts were analyzed.Results indicated that either L.plantarum or S.cerevisiae or co-fermentation could consume the polysaccharides of highland barley to provide energy,and dramatically increase the contents of total protein and polyphenol.Gas chromatography-mass spectrometry(GC-MS)analysis revealed that the presence of S.cerevisiae was critical for production of the pleasant flavors,especially for the ethyl ester substances including hexadecanoic acid ethyl,hexanoic acid ethyl ester and so on.Meanwhile,we found that fermented highland barley extracts by L.plantarum exhibited stronger lipid-lowering effects in Caenorhabditis elegans than that by S.cerevisiae,while the co-fermentation not only emitted pleasant odors but also exerted high hypolipidemic function.In all,co-fermentation by L.plantarum and S.cerevisiae was proposed to be a promising processing to improve the flavor and functional properties of highland barley.

Bioethanol Production from Rice Straw Enzymatically Saccharified by Fungal Isolates, Trichoderma viride F94 and Aspergillus terreus F98

Egypt faces a high population growth rate nowadays, which demands for an increase in agricultural production efficiency. Consequently, agricultural field residues will increase. Rice straw is one of the main agriculture residues in Egypt. So this study was performed on rice straw as a resource for production of bioethanol. Eight microbial isolates, five yeasts and three fungi were isolated from rice straw. Yeast isolates were selected for their ability to utilize different sugars and cellulose. Chipped and grinded rice straw was subjected to different pretreatment methods physically through steam treatment by autoclaving and different doses of gamma γ irradiation (50 and 70 Mrad). Autoclaved pretreated rice straw was further enzymatically treated throughout solid state fermentation process by different fungal isolates;F68, F94 and F98 producing maximum total reducing sugars of 12.62, 13.58, 17.00 g/L, respectively. Bioethanol production by separate microbial hydrolysis and fermentation (SHF) process of rice straw hydrolysate was performed by the two selected fungal isolates;Trichoderma viride F94 and Aspergillus terreus F98 and two yeast isolates (Y26 and Y39). The two yeast isolates have been identified by 18S, RNA as Candida tropicalis Y26 and Saccharomyces cerevisiae Y39. SHF processes by F94 and Y26 produced 45 gallon/ton rice straw while that of F98 and Y39 produced 50 gallon/ton rice straw.

基于Lachancea thermotolerans-Saccharomyces cerevisiae双酵母混合发酵酸啤酒的工艺研究

酸啤酒正在被越来越多的消费者接受,也正在成为啤酒企业多元化布局的方向之一。该研究在不添加外源风味物质的前提下,基于啤酒双酵母混合发酵体系,将耐热拉钱斯氏酵母(Lachancea thermotolerans)与酿酒酵母(Saccharomyces cerevisiae)组合按照10∶1同时接种进行啤酒发酵,耐热拉钱斯氏酵母的发酵过程可产生一定量的乳酸(1.5~1.8 g/L),降低酒液pH值(3.3~3.4),得到酸味柔和的新型酸啤酒。相比于单一菌株发酵,双酵母混合发酵酸啤酒体系的风味物质产量都有所提高,产生了更多乙酸异丁酯和乙酸异戊酯等酯类物质,增强了酸啤酒的水果香气,果香馥郁、爽口协调,对啤酒的质量产生了积极影响,并且减少了糖化锅酸化等工艺操作步骤,实现更快速的发酵和稳定的过程控制,促进了酸啤酒的大规模生产应用。

Use of radiation in strains of Saccharomyces cerevisiae:A new technique for industrial applications

During the industrial fermentation process in the production of fuel ethanol, yeasts are subject to several stressing conditions. The survival and the permanence of strains introduced in the process correlate with the capability of these yeasts in resisting to physical and chemical stresses, as well as their recovering ability to compete with contaminating micro-organisms commonly present in this industrial process. We aim at the selection of Saccharomyces cere visiae strains having this capability and ability. In this sense, cultivations of strains with industrial interest were irradiated with gammas ray at a wide dose interval. Growing curves for the strains were analyzed by means of their relative growth, a new concept here introduced, which allows a better understanding of the growing and recovering processes following radiative stress. It was found that gamma radiation could be used as an alternative method to quantify growing capabilities of S. cerevisiae strains under stressing conditions. It was also shown that this radiological method could be utilized as an additional procedure to select best robust industrial strains. This radiological method simplifies traditional analysis of strain viability, by avoiding the great number of necessary and consecutive fermentation assays.

酒精酵母Saccharomyces cerevisiae 4-S的高密度培养及其酒精发酵性能

实验比较了分批培养、分批补料及连续流加培养对酒精酵母细胞生长的影响。结果显示,分批补料流加或连续流加(流加速率0.133 g.L-1.m-1)有利于酒精酵母菌Saccharomyces cerevisiae4-S的快速生长和得到较高的细胞浓度,分批补料流加或连续流加培养44 h,发酵液中酒精酵母细胞浓度分别达到71.82(细胞干重-DCW)g.L-1和82.01(DCW)g.L-1,比分批培养分别提高了155.68%和191.95%。同时,对高密度酒精酵母的酒精发酵性能和循环利用研究结果显示,在细胞浓度为82.01(DCW)g.L-1,糖浓度为350 g.L-1,发酵36h发酵液中乙醇浓度达到16.23%,比普通酵母浓度28.09(DCW)g.L-1批发酵时间缩短12 h,乙醇浓度提高了24.4%,酵母的有效重复利用次数达4批次。

Increasing isobutanol yield by double-gene deletion of PDC6 and LPD1 in Saccharomyces cerevisiae

As a new biofuel, isobutanol has received more attentions in recent years. Because of its high tolerance to higher alcohols, Saccharomyces cerevisiae has potential advantages as a platform microbe to produce isobutanol. In this study, we investigated integration effects of enhancing valine biosynthesis by overexpression of ILV2 and BAT2 with eliminating ethanol formation by deletion of PDC6 and decreasing acetyl-Co A biosynthesis by deletion of LPD1 on isobutanol titers. Our results showed that deletion of LPD1 in strains overexpressing BAT2 and ILV2 increased isobutanol titer by 5.3-fold compared with control strain. Additional deletion of PDC6 in lpd1Δ strains carrying overexpressed BAT2 and ILV2 further increased isobutanol titer by 1.5 fold. Overexpression of BAT2 and ILV2 in lpd1Δ strains and pdc6Δ strains decreased ethanol titers. Glycerol titers of the engineered strains did not have greater changes than that of control strain, while their acetic acid titers were higher, perhaps due to the imbalance of cofactors in isobutanol synthesis. Our researches suggest that double-gene deletion of PDC6 and LPD1 in strains overexpressing BAT2 and ILV2 could increase isobutanol production dramatically than single-gene deletion of PDC6 or LPD1. This study reveals the integration effects of overexpression of ILV2/BAT2 and double-gene deletion of LPD1 and PDC6 on isobutanol production, and helps understanding future developments of engineered strains for producing isobutanol.