Process conditions for a rapid in situ transesterification for biodiesel production from oleaginous yeasts

BACKGROUND: Microbial oils produced by diverse microorganisms are being considered as alternative sources of triglycerides for biodiesel production. However, the standalone production of biodiesel from microorganisms is not currently economically feasible. In case of yeasts, the use of low-value nutrient sources in microbial production and the implementation of cost-efficient downstream processes could reduce costs and make microbial lipids competitive with other commodity-type oils in biodiesel production. Industrial biodiesel synthesis from oleaginous seeds is currently based on a multistep process. However, a simple process called in situ transesterification (ISTE), which takes place within the biomass without a previous lipid extraction step, is receiving increasing interest. In this work, the optimal conditions for an ISTE process to obtain biodiesel from previously selected oleaginous yeast (Rhodotorula graminis S1/S2) were defined using the response surface methodology (RSM). RESULTS: Using the RSM approach, the optimal conditions for the maximum yield with minimum reaction time included a methanol-to-biomass ratio of 60:1, 0.4 M H2SO4, and incubation at 70°C for 3 h. The optimized in situ process yield was significantly higher (123%) than that obtained with a two-step method in which fatty acids from saponifiable lipids were first extracted and then esterified with methanol. The composition of the fatty acid methyl ester mixture obtained from R. graminis S1/S2 by ISTE met Uruguayan standards for biodiesel. CONCLUSION: The characteristics achieved by the optimized method make microbial oil a potential alternative for biodiesel production from yeast at an industrial scale.

Saccharomyces cerevisiae populations and other yeasts associated with indigenous beers (chicha) of Ecuador

ABSTRACT Chicha, a type of beer made mainly with maize or cassava, is a traditional fermented beverage of the Andean region. There have only been a few studies on yeasts associated with chicha fermentation, and the species diversity occurring during the production of this beverage is not known. The objective of this study was to determine the biodiversity of yeasts in chicha, and to characterize the Saccharomyces cerevisiae populations associated with the production of chicha de jora, seven-grain chicha, chicha de yuca, and chicha de morocho in Ecuador. The molecular diversity of S. cerevisiae populations was determined by restriction polymorphism mitochondrial profiles. The beverages were characterized based on their physicochemical parameters. Twenty-six species were identified, and the most prevalent species were S. cerevisiae and Torulaspora delbrueckii. Other yeast species were isolated at low frequencies. Among 121 isolates of S. cerevisiae, 68 different mtDNA molecular profiles were identified. These results showed that chichas are fermented by a high number of different strains of S. cerevisiae. Some other species provided a minor contribution to the fermentation process. The chicha presented generally similar physicochemical parameters to those observed for other traditional fermented beverages, and can be considered as an acid fermented beverage.

Selection of starter cultures for the production of sour cassava starch in a pilot-scale fermentation process

ABSTRACT Sour cassava starch (Polvilho azedo) is obtained from a spontaneous fermentation conducted by microorganisms from raw materials and fermentation tanks. This product is traditionally used in the baking industry for the manufacture of biscuits and Brazilian cheese breads. However, the end of fermentation is evaluated empirically, and the process occurs without standardization, which results in products of inconsistent quality. Predominant microbiota from a cassava flour manufacturer was isolated in order to select starter cultures for the production of sour cassava starch in a pilot-scale fermentation process. Lactic acid bacteria and yeasts were isolated, enumerated and grouped by Restriction Fragment Length Polymorphism, and PCR fingerprinting, respectively. One isolate of each molecular profile was identified by sequencing of the rRNA gene. LAB were prevalent throughout the entire process. Lactobacillus brevis (21.5%), which produced the highest values of acidity, and Lactobacillus plantarum (13.9%) were among the most frequent species. Pichia scutulata (52.2%) was the prevalent yeast and showed amylolytic activity. The aforementioned species were tested as single and mixed starter cultures in a pilot-scale fermentation process for 28 days. L. plantarum exhibited better performance as a starter culture, which suggests its potential for the production of sour cassava starch.

The isolation of pentose-assimilating yeasts and their xylose fermentation potential

ABSTRACT For the implementation of cellulosic ethanol technology, the maximum use of lignocellulosic materials is important to increase efficiency and to reduce costs. In this context, appropriate use of the pentose released by hemicellulose hydrolysis could improve de economic viability of this process. Since the Saccharomyces cerevisiae is unable to ferment the pentose, the search for pentose-fermenting microorganisms could be an alternative. In this work, the isolation of yeast strains from decaying vegetal materials, flowers, fruits and insects and their application for assimilation and alcoholic fermentation of xylose were carried out. From a total of 30 isolated strains, 12 were able to assimilate 30 g L-1 of xylose in 120 h. The strain Candida tropicalis S4 produced 6 g L-1 of ethanol from 56 g L-1 of xylose, while the strain C. tropicalis E2 produced 22 g L-1 of xylitol. The strains Candida oleophila G10.1 and Metschnikowia koreensis G18 consumed significant amount of xylose in aerobic cultivation releasing non-identified metabolites. The different materials in environment were source for pentose-assimilating yeast with variable metabolic profile.

Effect of ultraviolet radiation on physiological and biochemical properties of yeast Saccharomyces cerevisiae during fermentation of ultradispersed starch raw material

Background: Study of correlation between pretreatment of yeast with ultraviolet radiation and efficiency of further fermentation of wort made of ultrafine grain particles to ethanol. Results: We investigated three races of industrial yeast Saccharomyces cerevisiae (native and irradiated by ultraviolet). Physiological properties during fermentation of starchy wort were tested in all variants. It was shown that activation of the yeast by ultraviolet radiation allows to further increase the ethanol yield by 25% on average compared with the native yeast races when using thin (up to micro- and nano-sized particles) or standard grain grinding. Conclusions: Using mechanical two-stage grinding of starchy raw materials and ultraviolet pretreatment of yeast, the efficiency of saccharification of starch and fermentation of wort to ethanol was increased.

Use of homologous recombination in yeast to create chimeric bovine viral diarrhea virus cDNA clones

Abstract The open reading frame of a Brazilian bovine viral diarrhea virus (BVDV) strain, IBSP4ncp, was recombined with the untranslated regions of the reference NADL strain by homologous recombination in Saccharomyces cerevisiae, resulting in chimeric full-length cDNA clones of BVDV (chi-NADL/IBSP4ncp#2 and chi-NADL/IBSP4ncp#3). The recombinant clones were successfully recovered, resulting in viable viruses, having the kinetics of replication, focus size, and morphology similar to those of the parental virus, IBSP4ncp. In addition, the chimeric viruses remained stable for at least 10 passages in cell culture, maintaining their replication efficiency unaltered. Nucleotide sequencing revealed a few point mutations; nevertheless, the phenotype of the rescued viruses was nearly identical to that of the parental virus in all experiments. Thus, genetic stability of the chimeric clones and their phenotypic similarity to the parental virus confirm the ability of the yeast-based homologous recombination to maintain characteristics of the parental virus from which the recombinant viruses were derived. The data also support possible use of the yeast system for the manipulation of the BVDV genome.

Isolation and characterization of yeasts from fermented apple bagasse as additives for ruminant feeding

Abstract Solid-state fermentation can be used to produce feeds for ruminants, which can provide an enriched population of yeasts to improve ruminal fermentation. Fermentation of apple bagasse was performed to obtain a yeast-rich product, with the objective of isolating, identifying, and characterizing yeast strains and testing their capability to enhance in vitro ruminal fermentation of fibrous feeds. Yeasts were isolated from apple bagasse fermented under in vitro conditions, using rumen liquor obtained from cannulated cows and alfalfa as a fibrous substrate. A total of 16 new yeast strains were isolated and identified by biochemical and molecular methods. The strains were designated Levazot, followed by the isolate number. Their fermentative capacity was assessed using an in vitro gas production method. Strain Levazot 15 (Candida norvegensis) showed the greatest increase in gas production (p < 0.05) compared with the yeast-free control and positively affected in vitro ruminal fermentation parameters of alfalfa and oat straw. Based on these results, it was concluded that the Levazot 15 yeast strain could be potentially used as an additive for ruminants consuming high-fiber diets. However, further studies of effects of these additives on rumen digestion, metabolism, and productive performance of ruminants are required.

Caracterización molecular y fisiológicas de levaduras de suelo trumao / Molecular and physiological characterization of yeasts of trumao soil

Las levaduras juegan un importante rol en la naturaleza siendo el mayor reservorio de ellas el suelo. Mediante el método de las diluciones seriadas y posterior siembra en agar Sabouraud se aislaron en cultivo puro 77 cepas de levaduras desde un mismo suelo trumao del sur de Chile, usado como pradera permanente (30 cepas), pradera en rotación (30 cepas) y como control bosque nativo (17 cepas), estas cepas se identificaron molecularmente por PCR-RFLP en conjunto con secuenciación del rDNA de ITS-5.8S, además se realizo una caracterización fisiológica (asimilación fuente de carbono, de nitrógeno y fermentación de azucares) a cada cepa. Mediante las técnicas moleculares las 77 cepas se reunieron en 10 grupos, de estos solamente tres grupos se pudieron identificar a nivel de especie y uno hasta género: Devariomyces hansenii. Pichia fermentan. Kazachstania exigua., Candida sp.

Yeasts plays an important role in nature, It is the largest reservoir of soil them. By the method of serial dilutions and subsequent planting in Sabouraud agar were isolated in pure culture 77 strains of yeast from the same volcanic ash soil of southern Chile, used as permanent pasture (30 strains), rotation pasture (30 strains) and native forest as a control (17 strains), these strains were identified molecularly by PCR-RFLP in conjunction with rDNA sequencing ITS-5.8S, physiological characterization addition was performed to each strain (carbon and nitrogen source assimilation and fermentation of sugars). Using molecular techniques met the 77 strains in 10 groups; only three groups could be identified to species level and one to gender: Devariomyces hansenii; Pichia fermented; Kazachstania exigua; Candida sp.

Ethanol production in Brazil: a bridge between science and industry

ABSTRACT In the last 40 years, several scientific and technological advances in microbiology of the fermentation have greatly contributed to evolution of the ethanol industry in Brazil. These contributions have increased our view and comprehension about fermentations in the first and, more recently, second-generation ethanol. Nowadays, new technologies are available to produce ethanol from sugarcane, corn and other feedstocks, reducing the off-season period. Better control of fermentation conditions can reduce the stress conditions for yeast cells and contamination by bacteria and wild yeasts. There are great research opportunities in production processes of the first-generation ethanol regarding high-value added products, cost reduction and selection of new industrial yeast strains that are more robust and customized for each distillery. New technologies have also focused on the reduction of vinasse volumes by increasing the ethanol concentrations in wine during fermentation. Moreover, conversion of sugarcane biomass into fermentable sugars for second-generation ethanol production is a promising alternative to meet future demands of biofuel production in the country. However, building a bridge between science and industry requires investments in research, development and transfer of new technologies to the industry as well as specialized personnel to deal with new technological challenges.

Isolation and characterization of phenol degrading yeasts from wastewater in the coking plant of Zarand, Kerman

Abstract Phenol and phenolic compounds are environmental pollutants present in industrial wastewaters such as coal tar, oil refineries and petrochemical plants. Phenol removal from industrial effluents is extremely important for the protection of environment. Usually, phenol degradation is carried out by physicochemical methods that are costly and produce hazardous metabolites. Recently, phenol biodegradation has been considered. Yeasts are the most important phenol biodegraders. In this study, the phenol-degrading yeast from environmental samples (soil and wastewater) was isolated from the coking plant of Zarand, Kerman. Then total heterotrophic yeasts were counted. The soil samples had higher rates of yeast degrader, in comparison to wastewater samples. After three passages, four yeasts (K1, K2, K7 and K11) that had the highest growth rate were selected for further study. Also, these yeasts were able to remove phenol measured by Gibbs reagent. The effect of four different concentrations of phenol (50, 125, 200 and 275) mg L−1 was measured and three degradation patterns in these yeasts were observed. The hydrophobicity and emulsification activity were measured in all eleven yeasts. Finally, strong yeasts in phenol degrading yeasts were identified by molecular method using amplification of 18S rRNA gene region. The sequencing results showed that these isolated yeasts belonged to Candida tropicalis strain K1, Pichia guilliermondii strain K2, Meyerozyma guilliermondii strain K7 and C. tropicalis strain K11.

Energia metabolizável da levedura de cana-de-açúcar submetida a diferentes temperaturas e tempos de secagem para frangos de corte e poedeiras / Metabolizable energy of sugar cane yeast submitted to different temperatures and drying times for broilers and layers hens

Objetivou-se determinar a temperatura e o tempo de secagem por rolos rotativos, aos quais a, levedura de cana-de-açúcar é submetida que permitam seu melhor aproveitamento energético por galinhas poedeiras e frangos de corte. Para isso foram realizados três ensaios de metabolismo para determinar os valores de energia metabolizável aparente (EMA), aparente corrigida para nitrogênio (EMAn) e os coeficientes de metabolizabilidade aparente da matéria seca (CMMS) e da energia bruta (CMEB). O primeiro ensaio foi conduzido com galinhas poedeiras (E1), o segundo com frangos de corte (E2) em crescimento e o terceiro com frangos de corte em diferentes idades (E3)...

This study aimed to determine the temperature and drying time through rotative rolls, that sugar cane yeast is subjected to in order to allow best energy utilization by laying hens and broilers. Three metabolism trials were conducted to determine the values of apparent metabolizable energy (AME) and apparent corrected for nitrogen balance (AMEn), coefficient of apparent metabolizable dry matter (CAMDM) and gross energy (CAMGE). The first experiment was conducted with laying hens (E1), the second with broilers (E2) in growth and the third with broilers at different ages (E3)...

The ICY1 gene from Saccharomyces cerevisiae affects nitrogen consumption during alcoholic fermentation

Background Saccharomyces cerevisiae is the main microorganism responsible for alcoholic fermentation. In this process, the consumption of nitrogen is of great importance since it is found in limiting quantities and its deficiency produces sluggish and/or stuck fermentations generating large economic losses in the wine-making industry. In a previous work we compared the transcriptional profiles between genetically related strains with differences in nitrogen consumption, detecting genes with differential expression that could be associated to the differences in the levels of nitrogen consumed. One of the genes identified was ICY1. With the aim of confirming this observation, in the present work we evaluated the consumption of ammonium during the fermentation of strains that have deleted or overexpressed this gene. Results Our results confirm the effect of ICY1 on nitrogen uptake by evaluating its expression in wine yeasts during the first stages of fermentation under low (MS60) and normal (MS300) assimilable nitrogen. Our results show that the mRNA levels of ICY1 diminish when the amount of assimilable nitrogen is low. Furthermore, we constructed strains derived from the industrial strain EC1118 as a null mutant in this gene as well as one that overexpressed it. Conclusions Our results suggest that the expression of ICY1 is regulated by the amount of nitrogen available in the must and it is involved in the consumption of ammonium, given the increase in the consumption of this nitrogen source observed in the null mutant strain.

The diversity, extracellular enzymatic activities and photoprotective compounds of yeasts isolated in Antarctica

The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island) and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano) soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia), Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7 percent produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4ºC and 20ºC, indicating that they could be metabolically active in the sampled substrates.

Leveduras no rúmen de caprinos e bovinos de corte criados em pastagem tropicais / Yeasts in the rumen of beef steers and goats raised on tropical pasture

Yeasts suplemented in the rumen have been produced benefic interations in the digestion and in the health of the ruminants. This study aimed to quantify, to isolate and, to identify aerobic fungi and yeasts naturally present in the rumen of goats and cattle raised on tropical pastures of the North of Minas Gerais State, Brazil. Samples of 15mL of ruminal juice from 18 hibrid goats and 31 crossbred Nellore steers were used. The physico-chemical characteristics of the samples were evaluated and mycological culture, quantification, and identification of the aerobic fungi were performed. The results indicated the absence of yeasts in the ruminal fluid of steers. However, theses microorganisms were cultured from ruminal juice for all evaluated goats, at an average concentration of 3.2 x 10VCFU/mL. The species Pichia membranifaciens was the most frequently identified yeast, suggesting its participation in the ruminal microbiot of theses small ruminants.

High density process to cultivate Lactobacillus plantarum biomass using wheat stillage and sugar beet molasses

Background: Owing to the growing interest in biofuels, the concept of a biorefinery where biomass is converted to a variety of useful products is gaining ground. We here present how distillery waste is combined with a by-product from a sugar production, molasses, to form a medium for the growth of Lactobacillus plantarum with yields and biomass densities comparable with conventional industrial media. Such approach enables a cost-effective utilization of the problematic wastewater from ethanol and a by-product from sugar production. It is the first approach that attempts to find low-cost media for the production of Lactobacillus plantarum biomass. Results: This study suggests that sieved wheat stillage enriched by adding 1.77 g/l yeast extract and 10 percent molasses (v/v), with NH4OH used for pH adjustment, may be used as a media for large-scale cultivation of L. plantarum. Such composition of the medium permits a high density of lactic acid bacteria (1.6 x 10(10) cfu/ml) to be achieved. Conclusions: The use of a fermentation medium consisting of distillery wastewater and molasses to obtain value-added products (such as LAB biomass and lactic acid) is a possible step for classical ethanol production to move towards a biorefinery model production in which all by and waste products are utilized to increase produced values and reduce waste production. This enables a cost-effective utilization of the problematic wastewater from ethanol and sugar production.

Optimization of killer assays for yeast selection protocols / Optimización de la actividad killer para protocolos de selección de levaduras

A new optimized semiquantitative yeast killer assay is reported for the first time. The killer activity of 36 yeast isolates belonging to three species, namely, Metschnikowia pulcherrima, Wickerhamomyces anomala and Torulaspora delbrueckii, was tested with a view to potentially using these yeasts as biocontrol agents against the wine spoilage species Pichia guilliermondii and Pichia membranifaciens. The effectiveness of the classical streak-based (qualitative method) and the new semiquantitative techniques was compared. The percentage of yeasts showing killer activity was found to be higher by the semiquantitative technique (60%) than by the qualitative method (45%). In all cases, the addition of 1% NaCl into the medium allowed a better observation of the killer phenomenon. Important differences were observed in the killer capacity of different isolates belonging to a same killer species. The broadest spectrum of action was detected in isolates of W. anomala NPCC 1023 and 1025, and M. pulcherrima NPCC 1009 and 1013. We also brought experimental evidence supporting the importance of the adequate selection of the sensitive isolate to be used in killer evaluation. The new semiquantitative method proposed in this work enables to visualize the relationship between the number of yeasts tested and the growth of the inhibition halo (specific productivity). Hence, this experimental approach could become an interesting tool to be taken into account for killer yeast selection protocols.

En este trabajo se presenta un nuevo ensayo semicuantitativo que optimiza la detección de actividad killer en levaduras. Se evaluó la actividad killer de 36 cepas pertenecientes a las especies Metschnikowia pulcherrima, Wickerhamomyces anomala y Torulaspora delbrueckii, en vista del potencial uso de estas levaduras como agentes de biocontrol frente a las especies contaminantes de vinos Pichia guilliermondii y Pichia membranifaciens. Se comparó la efectividad de la técnica clásica basada en estrías (método cualitativo) con la del nuevo método semicuantitativo. El porcentaje de levaduras que mostraron actividad killer fue más alto cuando se utilizó el método semicuantitativo (60%) que con el método cualitativo (45%). En todos los casos, el agregado de 1% de NaCl en el medio permitió una mejor observación del fenómeno killer. Se observaron importantes diferencias en la capacidad killer de diferentes cepas dentro de la misma especie. Se detectaron dos cepas de W. anomala (NPCC 1023 y 1025) y dos cepas de M. pulcherrima (NPCC 1009 y 1013) con un amplio espectro de acción, ya que fueron capaces de inhibir el desarrollo de las tres levaduras sensibles evaluadas. La evidencia experimental demuestra la importancia de una adecuada selección de la cepa sensible al evaluar la actividad killer. El nuevo método semicuantitativo propuesto en este trabajo permite visualizar la relación entre el número de levaduras sembradas y el halo de inhibición del crecimiento (productividad específica). En conclusión, este método resulta una herramienta interesante para ser tenida en cuenta en los protocolos de selección de levaduras killer.

Kinetic characteristics of beta-cyclodextrin production by cyclodextrin glycosyltransferase from newly isolated Bacillus sp. C26

The kinetic characteristics of beta-cyclodextrin production by a cyclodextrin glycosyltransferase (CGTase) produced by Bacillus sp. C26, a new isolate from a soil sample was investigated. Considering highest yield and initial production rate of beta-cyclodextrin, among the starches examined, soluble starch, tapioca starch, sago starch, corn starch and rice starch, tapioca starch was the best substrate for this CGTase. The optimum temperature for tapioca starch gelatinization prior to its use as a substrate for beta-cyclodextrin production was 65ºC. The yield and initial production rate of beta-cyclodextrin increased with increasing starch concentration up to 6 percent and an enzyme concentration up to 48 U/g-starch. The kinetic parameters of Vmax and Km of beta-cyclodextrin production from tapioca starch by CGTase were 1.59 mg/mL/h and 22.3 mg/mL, respectively. Considering high initial production rate and high yield of beta-cyclodextrin, the optimum reaction temperature was at 50ºC. This study provided the necessary kinetic information that may be useful to define the most suitable condition for industrialized production of beta-cyclodextrin with the high yield and productivity.

Proteins in a DNA world: expression systems for their study / Proteínas en el mundo del DNA: Sistemas de expresión para su estudio

Every day, new proteins are discovered and the need to understand its function arises. Human proteins have a special interest, because to know its role in the cell may lead to the design of a cure for a disease. In order to obtain such information, we need enough protein with a high degree of purity, and in the case of the human proteins, it is almost impossible to achieve this by working on human tissues. For that reason, the use of expression systems is needed. Bacteria, yeast, animals and plants have been genetically modified to produce proteins from different species. Even "cell-free" systems have been developed for that purpose. Here, we briefly review the options with their advantages and drawback, and the purification systems and analysis that can be done to gain understanding on the function and structure of the protein of interest.

Cada día, nuevas proteínas son descubiertas y surge la necesidad de caracterizarlas, siendo las de origen humano las que presentan un mayor interés. Conocer su función nos ayudará a entender padecimientos y diseñar una posible cura. Sin embargo, obtener suficiente cantidad de proteínas humanas en cantidad para llevar a cabo los análisis pertinentes, presenta una gran dificultad. Por tal razón, es necesario el uso de sistemas de expresión de proteínas heterólogas. Bacterias, levaduras, animales y plantas han sido modificados genéticamente para expresar proteínas de otras especies, e incluso sistemas in vitro han sido desarrollados para producir proteínas. En este artículo se revisan brevemente las opciones con sus ventajas y desventajas, así como las estrategias de purificación y los análisis que se pueden llevar a cabo para avanzar en el conocimiento de la función y estructura de la proteína de interés.

Citric acid production from glucose by yeast Candida oleophila ATCC 20177 under batch, continuous and repeated batch cultivation

The effect of air saturation on citric acid production in batch, repeated batch and chemostat cultures has been studied. It was shown that, under continuous fermentation (chemostat mode), the highest concentration of citric acid equal of 98 g/l was produced at 20% of air saturation. In contrary to continuous fermentation, displaying an optimum at 20%, 80% air saturation yielded higher values in repeated batch fermentation process. 167 g/l citric acid were produced continuously with the fill and drain technique at 4.85 days, at 80% air saturation, compared with 157.6 g/l achieved within 5.4 days at 20%. Under repeated batch fermentation, the formation rate of the generic product (Rj) as well as the specific citric acid productivity (mp) reached a maximum of 1.283 g/(l x hr) at 4.01 days and of 0.0375 g/(g x hr) at 4.58 days, respectively. The glucose consumption rate (Rx) reached a maximum value of 3.33 g/(l x hr) entering stationary phase after 2.56 days at a glucose concentration of 131.2 g/l.

Improvement of SCP production and BOD removal of whey with mixed yeast culture

This research emphasizes on single cell protein (SCP) production and Biochemical Oxygen Demand (BOD) removal from whey with mixed yeast culture. For this purpose, 11 yeast strains were isolated from dairy products (M1-M11) and the strains were identified by morphological and physiological properties. These yeast strains were tested for their ability to reduce the BOD and to produce SCP from whey. Among these strains, K. lactis (M2) had the most SCP production from whey with the yield of 11.79 g/l. Ammonium sulphate as nitrogen source had an increasing effect on biomass yield. The mixed culture of the isolated yeast strains with Saccharomyces cerevisiae was used in order to increase the biomass yield and BOD removal. The highest biomass yield (22.38 g/l) and reduction of initial BOD from 30000 to 3450 mg/l were obtained with the mixed culture of K. lactis (M2) and S. cerevisiae.