Influence of specific growth rate on biomass yield, productivity, and compostion of Candida utilis in batch and continuous culture.

Candida utilis was grown in batch and continuous culture on prickly pear juice as sole carbon and energy source. In batch culture the maximum specific growth rate (mum) and the substrate yield coefficient (Yps) varied according to sugar concentration. When the fermentation was carried out with 1% sugar, mum and Ys were 0.47/h and 42.6%, respectively. The best yields occurred in a chemostat at the pH range of 3.5 to 4.5 and temperature of 30 C. A beneficial effect on Ys was observed when the dilution rate (D) was increased. At a D of 0.55/h, the productivity was 2.38 g/liter per h. The maintenance coefficient attained a value of 0.09 g of sugar/g of biomass per h. Increases of D produced higher protein contents of the biomass. The information obtained indicates that protein production with Candida utilis, using prickly pear juice, should be carried out a high dilution rates where the Ys and protein content of the cell mass are also higher.     

Continuous ethanol production from Jerusalem artichoke tubers. I. Use of free cells of Kluyveromyces marxianus

The Continuous fermentation of Jerusalem artichoke juice to ethanol by free cells of Kluyveromyces marxianus UCD (FST) 55-82 has been studied in a continuous-stirred-tank bioreactor at 35 degrees C and pH 4.6. A maximum yield of 90% of the theoretical was obtained at a dilution rate of 0.05 h(-1). About 95% of the sugars were utilized at dilution rates lower than 0.15 h(-1). Volumetric ethanol productivity and volumetric biomass productivity reached maximum values of 7 g ETOH/L/h and 0.6 g dry wt/L/h, respectively, at a dilution rate of 0.2 h(-1). The maintenance energy coefficient for K. marxianus culture was found to be 0.46 g sugar/g biomass/h/ Oscillatory behavior was following a change in dilution rate from a previous steady state and from batch to continuous culture. Values of specific ethanol production rate and specific sugar uptake were found to increase almost linearly with the increase of the dilution rate. The maximum specific ethanol production rate and maximum specific sugar uptake rate were found to be 2.6 g ethanol/g/ cell/h and 7.9 sugars/g cell/h, respectively. Washout occurred at a dilution rate of 0.41 h(-1).     

Use of waste Chinese cabbage as a substrate for yeast biomass production

The possibility of using waste Chinese cabbage (Brassica campestris) as a substrate for microbial biomass production was investigated. The juice from waste Chinese cabbage contains relatively high amounts of reducing sugars suitable for yeast culture. The cell mass and protein content of four species of yeast, Candida utilis, Pichia stipitis, Kluyveromyces marxianus, and Saccharomyces cerevisiae, were determined when cultured in juice extracted from cabbage waste. Compared to YM broth containing the same level of sugar, all the strains except C. utilis showed higher total protein production in cabbage juice medium (CJM).     

Continuous growth kinetics of Candida utilis in pineapple cannery effluent

Candida utilis was grown on a pineapple cannery effluent as the sole carbon and energy source in a chemostat at dilution rates between 0.10 and 0.62 h(-1) to determine the growth kinetics. The principal sugars in the effluent were sucrose, glucose, and fructose. The cell yield coefficient on carbohydrate varied with dilution rate and a maximum value of 0.63 was observed at a dilution rate of 0.33 h(-1). The steady-state concentrations of carbohydrate, reducing sugar, and chemical oxygen demand (COD) appeared to follow Monod saturation kinetics with increasing dilution rate, although none of the measured parameters represented a pure substrate. The maximum specific growth rate and reducing sugar saturation constant were 0.64 h(-1) and 0.060 g/L, respectively. A maximum cell mass productivity of 2.3 g/L h was observed at a dilution rate of 0.51 h(-1). At this dilution rate, only 68% of the COD was removed. A 95% COD removal was attained at a dilution rate of 0.10 h(-1). Optimal yeast productivity and COD reduction occurred at a dilution rate of 0.33 h(-1).     

Effect of elevated temperature on the morphology of a chemostat culture of Candida utilis yeasts and on their content of nucleic acids

The effect of elevated temperature (40 degrees C) on a chemostat culture of Candida utilis was studied at different rates of dilution, D = 0.1 and 0.3 hr-1. The cells in the fermenter being in the stationary state at the optimum temperature of 30 degrees C were gradually washed-out 5 hours after the action of this temperature, and the population consisted of non-divided cells. In the majority of such "double" cells, the nucleus was contained in both the parent and the daughter parts. The content of RNA decreased by 31%, that of DNA, by 20%, and that of protein, by 13% (per 1 mg of biomass).     

Two-stage continuous fermentation of Saccharomycopsis fibuligeria and Candida utilis

Biomass production and carbohydrate reduction were determined for a two-stage continuous fermentation process with a simulated potato processing waste feed. The amylolytic yeast Saccharomycopsis fibuligera was grown in the first stage and a mixed culture of S. fibuligera and Candida utilis was maintained in the second stage. All conditions for the first and second stages were fixed except the flow of medium to the second stage was varied. Maximum biomass production occurred at a second stage dilution rate, D(2), of 0.27 h (-1). Carbohydrate reduction was inversely proportional to D(2), between 0.10 and 0.35 h (-1).     

响应面法优化产朊假丝酵母培养及干燥工艺

本研究旨在优化产朊假丝酵母液体培养参数及干燥保护剂配方,提高产朊假丝酵母液体培养数量,制备出高复苏率的活性干酵母。以装液量、转速、温度和培养时间为影响因素,设计单因素试验,并采用Box-Behnken试验设计及响应面分析法优化产朊假丝酵母液体培养方案。在此条件下培养酵母,以L-谷氨酸钠、乳糖、脱脂奶粉为影响因素进行单因素保护试验,并采用响应面分析法优化干燥保护剂的组合。结果表明,产朊假丝酵母最佳培养条件为装液量45 mL/250 m L,转速200 r/min,温度30℃,培养时间24 h。在此条件下,培养的产朊假丝酵母数量可以达到9.34×10~8CFU/mL;最佳保护剂组合1%L-谷氨酸钠、12%脱脂奶粉、6%乳糖,经干燥后,产朊假丝酵母复苏率达到81.9%。此时活菌数7.65×10~8CFU/g,比未加保护剂组提高了3.83倍。     

Kinetics of batch single cell protein production from rice polishings with Candida utilis in continuously aerated tank reactors

Single cell protein was produced from the defatted rice polishings by fermentation with Candida utilis in an aerated 14-L fermentor to optimize bioprocess variables. Maximum values of specific growth rate coefficient (mu, h(-1)), cell mass yield (Y(X/S), g/g) and cell mass productivity (g/Lh) were 0.31, 0.65, and 1.24, respectively under optimized conditions of aeration rate (1 v.v(-1) m(-1)), dissolved oxygen (50%), corn steep liquor (5%), temperature (35 degrees C), and substrate concentration (90 g rice polishings/L) in yeast salt medium (pH 6.0). The kinetic parameters for 50-L fermentor under same conditions were 0.33 h(-1), 0.66 g/g, 1.33 g/Lh, 2.25 g/Lh, 1.23 g/Lh, 0.45 g/g substrate and 0.20 g/g cell h for mu, Y(X/S), Q(X), Q(S), Q(CP), Y(TP/S), and q(CP), respectively and were significantly higher than their respective values reported on C. utilis in batch culture studies. This biomass protein contained 23.6%, 32.75%, 11.50%, 12.95%, 10.5%, and 0.275% true protein, crude protein, crude fiber, ash, cellulose and RNA content respectively. This implied that the fermentation process could be up scaled to manufacture animal feed. Gross metabolizable energy content of dried SCP was 29,711 kcal/kg and indicated that the SCP could serve both as energy as well as a protein source. Yeast can replace expensive feed ingredients currently being incorporated in poultry feed and can reduce cost of poultry ration by 0.33 US dollars-0.51 US dollars/100 kg bag and improve the economics of feed production in our country.     

培养方式对富硒产朊假丝酵母性能的影响

在摇瓶和5 L发酵罐水平上分别考察亚硒酸钠浓度及其添加方式对高性能(高有机硒含量和高谷胱甘肽含量)富硒产朊假丝酵母制备的影响.结果表明:亚硒酸钠添加质量浓度为15 mg/L时,产朊假丝酵母具有较好的富硒效果,但一次性添加对酵母细胞有较大的毒害作用.采用分批次添加亚硒酸钠的方法获得了较好的制备高性能富硒产朊假丝酵母的培养方式:发酵起始添加L-蛋氨酸10 mmol/L,并在发酵过程的12和15 h分别添加亚硒酸钠10和5 mg/L.在此培养方式下,产朊假丝酵母胞内谷胱甘肽和有机硒含量分别达到172.3 mg/L和1194 μg/g.     

Biomass production from glutamate fermentation wastewater by the co-culture of Candida halophila and Rhodotorula glutinis

In this study, the biomass production and pollutant removal from high-strength glutamate fermentation wastewater (GFW) using yeast isolates was investigated. Following enrichment culture, two species of yeasts, Candida halophila and Rhodotorula glutinis, were isolated from raw GFW with chemical oxygen demand (COD) and ammonia-nitrogen levels of 40 and 16 g l(-1), respectively. The binary mixed yeast culture was cultivated batchwise in 2.5-fold diluted GFW from which 85% of COD and 96% of reducing sugar were removed. The resulting yeast biomass contained 56% crude protein, 36.0% carbohydrate and 0.4% crude lipid. The amino acid composition of mixed yeast cells was balanced and was comparable with that of C. utilis and soybean.     

The use of a mixed culture of Geotrichum candidum,Candida krusei and Hansenula anomala for microbial protein production from whiskey distillery spent wash

Summary The use of a mixed culture of Geotrichum candidum, Hansenula anomala and Candida krusei for microbial protein production and treatment of whiskey distillery spent wash was investigated in both batch and continuous culture. Although capable of assimilating the same substrate constituents in pure culture the organisms used different constituents for growth in mixed continuous culture, allowing a stable mixed population to be established. The relative proportions of the three organisms in the population was dependent on the dilution rate.The mixed culture did not give superior yields or productivities, but did give proportionally greater COD reduction and would be expected to bemore stable and resistant to contamination.At a dilution rate of 0.10 h^–1 the mixed culture gave a biomass yield of 4.8 g l^–1, containing 55% crude protein, with a COD reduction of 31.5%, while in mixed batch culture a biomass yield of 12.5 g l^–1, containing 48% crude protein, was obtained with a COD reduction of 54.9%.     

Ethanol from whey: Continuous fermentation with cell recycle

The production of ethanol from cheese whey lactose has been demonstrated using a single-stage continuous culture fermentation with 100% cell recycle. In a two-step process, an aerobic fed batch operation was used initially to allow biomass buildup in the absence of inhibitory ethanol concentrations. In the anaerobic ethanol-producing second step, a strain of Kluyveromyces fragilis selected on the basis of batch fermentation data had a maximum productivity of 7.1 g ethanol/L/h at a dilution rate of 0.15 h(-1), while achieving the goal of zero residual sugar concentration. The fermentation productivity diminished when the feed sugar concentation exceeded 120 g/L despite the inclusion of a lipid mixture previous shown to enhance batch fermentation productivities.     

Growth characteristics of Candida utilis in a multistage culture system

The effect of increasing ethanol concentration in the feed on the growth and physiological activity of the yeast Candida utilis was studied. At steady-state of continuous culture with constant values of dilution rate, temperature, and pH in all fermenters in series biomass, ethanol and volatile acid concentrations, biomass yield and productivity and respiration activity were measured. In the three-stage system the maximum biomass concentration in the effluent and maximum productivity was achieved between 20 and 25 g ethanol/l in the feed. At higher concentrations, ethanol negatively affects the coupling of oxidative phosphorylation and respiratory control of cells resulting in a decrease in biomass yield and intracellular protein content. It was shown that the presence of acetate in the medium inhibits the respiration activity of yeasts growing on ethanol.     

Optimizing the continuous production of Candida utilis and Saccharomycopsis fibuliger on potato processing wastewater.

The yeasts Candida utilis and Saccharomycopsis fibuliger were propagated as a source of single-cell protein in a continuous, mixed, aerobic, single-stage cultivation on blancher water generated during potato processing. A series of steady-state experiments based on a two-level factorial design, half-replicate modified with an intermediate experiment, was performed to determine the effect of pH, 3.8 to 4.8; dissolved oxygen, 42 to 80% saturation; dilution rate, 0.17 to 0.31 h(-1); and temperature, 27 to 32 degrees C on the amount of carbon consumed, the rate of carbon consumption (R(c)), the amount of reducing sugar consumed, the rate of sugar consumption (R(g)), the amount of protein produced, the rate of protein production (R(p)), the yield from carbon, and the yield from reducing sugar. The results were analyzed by stepwise multiple regression and Fisher's least significant difference test. Analyses showed that high dilution rates resulted in increased R(c), R(g), and R(p) and indicated that a rate of 0.31 h(-1) was below the critical dilution rate. A temperature of 32 degrees C increased the amount of carbon consumed by 34%. A pH of 4.3 to 4.8 increased the amount of protein produced. The yield from carbon was constant, and the relatively high yield from reducing sugar indicated that other substrates were consumed. Dissolved oxygen was in excess at 42% saturation and above. Since C. utilis predominated the mixed cultures and amylase production appeared to be limited, a single-stage fermentation lacked efficiency. The experimental design allowed preliminary optimization of major environmental variables with relatively few experiments and provided a basis for future kinetic studies.     

Biomass production of yeast isolate from salad oil manufacturing wastewater

Conversion of oil-rich salad oil manufacturing wastewater (SOMW) into protein source for animal feed through biomass production of yeast isolate was investigated in this study. Five species of yeasts, including Rhodotorula rubra, Candida tropicalis, C. utilis, C. boidinii, Trichosporon cutaneum, were isolated from SOMW following enrichment culture. Of them, C. utilis was chosen as the sole biomass producer in the study due to its greatest oil uptake rate, 0.96 kg oil kg(-1) biomass d(-1), and highest specific growth rate, 0.25 h(-1). The cells of C. utilis contained 26% protein, 9% crude lipid, 55% carbohydrate and balanced amino acid compositions. The initial N:C ratio in SOMW drastically influenced oil reduction efficiency, biomass production and protein content of C. utilis, and therefore a range between 1:6 and 1:8 was recommended in consideration of these three factors simultaneously.     

High-cell-density cultivation of yeasts on disaccharides in oxygen-limited batch cultures

Many facultatively fermentative yeast species exhibit a "Kluyver effect": even under oxygen-limited growth conditions, certain disaccharides that support aerobic, respiratory growth are not fermented, even though the component monosaccharides are good fermentation substrates. This article investigates the applicability of this phenomenon for high-cell-density cultivation of yeasts. In glucose-grown batch cultures of Candida utilis CBS 621, the onset of oxygen limitation led to alcoholic fermentation and, consequently, a decrease of the biomass yield on sugar. In maltose-grown cultures, alcoholic fermentation did not occur and oxygen-limited growth resulted in high biomass concentrations (90 g dry weight L(-1) from 200 g L(-1) maltose monohydrate in a simple batch fermentation). It was subsequently investigated whether this principle could also be applied to Kluyveromyces species exhibiting a Kluyver effect for lactose. In oxygen-limited, glucose-grown chemostat cultures of K. wickerhamii CBS 2745, high ethanol concentrations and low biomass yields were observed. Conversely, ethanol was absent and biomass yields on sugar were high in oxygen-limited chemostat cultures grown on lactose. Batch cultures of K. wickerhamii grown on lactose exhibited the same growth characteristics as the maltose-grown C. utilis cultures: absence of ethanol formation and high biomass yields. Within the species K. marxianus, the occurrence of a Kluyver effect for lactose is known to be strain dependent. Thus, K. marxianus CBS 7894 could be grown to high biomass densities in lactose-grown batch cultures, whereas strain CBS 5795 produced ethanol after the onset of oxygen limitation and, consequently, yielded low amounts of biomass. Because the use of yeast strains exhibiting a Kluyver effect obviates the need for controlled substrate-feeding strategies to avoid oxygen limitation, such strains should be excellently suited for the production of biomass and growth-related products from low-cost disaccharide-containing feedstocks. (c) 1996 John Wiley & Sons, Inc.     

Evaluation of Escherichia coli and Candida utilis as a model continuous, competitive mixed culture system

The yeast Candida utilis and the bacterium Escherichia coli B/r were evaluated as a candidate experimental, continous, competitive mixed culture system under ammonia-nitrogen limited conditions at 30 degrees C. High dilution rates favored yeast dominance, while low dilution rates favored bacterial dominance. The hydrogen ion concentration was also demonstrated to be an effective manipulative variable for control of the yeast-bacterial mixed culture. Through trial-and-error manipulation of the pH for the mixed culture operating at constant dilution rate, it was possible to locate a metastable equilibrium point and to operate in the vicinity of that point for more than 24h. The reproducible emergence of a variant E. coli was also noted in this study.     

Utilization of statistics and experimental design in data collection and analysis

Application of experimental design techniques to Pirt's yield model shows that it is important to collect data at the lowest and highest specific growth rates. In the fed-batch fermentation process, values of specific growth rate can be varied from the maximum value at the start of the process to very low values near the end of the experiment. Candida utilis was cultivated using batch followed by fed-batch culture with glucose as the main source of carbon and energy. Values of substrate concentration, oxygen consumption, carbon dioxide evolution, liquid volume, flow rate cell concentration, and nitrogen concentration, which was an indirect measure of biomass, were measured. Least-squares estimates of the true biomass energetic yield and maintenance coefficient were obtained using a multivariate statistical analysis procedure referred to as the covariate adjustment procedure. Methods of selecting the best estimates using covariate adjustment are illustrated. The results show that useful parameter estimates with relatively short confidence intervals can be obtained using these statistical methods.     

Influence of glycerol production on the aerobic and anaerobic growth of the wine yeast Candida stellata

Candida stellata is frequently found in wine fermentations and may be used as a yeast starter in beverage production. In order to acquire additional knowledge on the physiology of C. stellata, a study on sugar metabolism in aerobic and anaerobic conditions was carried out. We found that under anaerobic conditions the low growth rate and biomass yield of C. stellata were due to the diversion of carbon flux from ethanol to glycerol. C. stellata had lower ADHI (alcohol dehydrogenase) activity (3-4 fold) and higher GPDH (glycerol-3-phosphate dehydrogenase) activity (40 and 15 times higher in anaerobiosis and aerobiosis respectively) than that of a Saccharomyces cerevisiae control strain. In aerobic sugar-limited chemostat culture C. stellata exhibited lower maximum biomass concentration [5.23 gl(-1) (dry weight)] than other respirofermentative yeasts at very low dilution rates (up to D = 0.042 h(-1)). While glycerol was constantly produced, ethanol and sugar residue appeared at D = 0.042 h(-1) and D = 0.065 h(-1) respectively. The tendency of C. stellata to form glycerol is probably the main cause of its very low growth and fermentation rates.     

A model for continuous fermentations with amylolytic yeasts

A two-stage, associative fermentation process is more effective for continuous yeast biomass production from starch than a single-stage mixed culture fermentation process. By operating two stages, competition for the same growth limiting substrate is reduced leading to efficient starch utilization. In this article, a mathematical model has been proposed for continuous, two-stage fermentation with a pure culture, amylolytic yeast in the first stage and a mixed culture second stage with a faster growing, nonamylolytic yeast. The model parameters were determined for Saccharomycopsis fibuligera and Candida utilis in continuous, single-stage, pure cultures. In the two-stage model, the effects of changes in dilution rate on biomass, amylase, reducing sugar, and starch concentration, and ratio of stage volumes on microbial composition are discussed and compared with experimental data.