Chitosan-coated paper packaging for specialty coffee beans: Coating characterization, bean and beverage analysis.

Cellulose-based packaging has received great attention due to its characteristics of biodegradability, sustainability, and recyclability. Natural polymer coatings are usually applied to the paper surface to enhance the barriers to water vapour and improve the mechanical properties. A chitosan-based coating for paper packaging was developed in this work to store specialty roasted coffee beans, evaluating two samples of chitosan (Sigma® and molasses chitosan), and following the physico-chemical and microbiological characteristics of coffee beans along a period of 60 days. Sensory tests (Ranking Descriptive Analysis and Preference Test) were applied to the beverage prepared with the roasted and ground coffee beans stored in each packaging. Thin chitosan films provided good coverage and adhesion on the paper. Improved mechanical properties and lower water permeability were observed in the chitosan-coated papers. The physicochemical and microbiological characteristics of the coffee beans were not influenced by the packaging along 60 days of storage. The molasses chitosan coating resulted in slightly darker roasted beans. In sensory evaluation, there is a clear difference between the chitosan samples, so that molasses chitosan-coated packaging had higher scores compared to Sigma® chitosan treatment for flavor and global impression in the preference analysis of the beverage. The molasses chitosan-coated packaging had three to four more consumers attributing the highest scores for the beverage prepared with the roasted beans stored in this type of packaging.

Maize treatment with yeast cells induces resistance against Fusarium rot.

Maize is one of the most cultivated species and requires agrochOPemicals for nutrition and pathogen control. Fusarium verticillioides is responsible for damaging plants and stored grains. Plants naturally exposed to stresses have defense mechanisms that are triggered by chemical or biological agents, known as induced systemic resistance. In this study, the yeast Torulaspora globosa (strain CCA6S01) was evaluated as an immune response promoter in maize against F. verticillioides. The treatments started 4 days after maize emergence and consisted of control (saline solution), acetylsalicylic acid (ASA, 100 mg/L), yeast cells (1 × 105 cells/mL), or yeast metabolites (cell-free filtrates). After running the treatments, the plants were inoculated with 1 mL of a F. verticillioides suspension at a concentration of 1 × 106 spores/mL. The application of yeast cells provided similar results to ASA treatment, a known inducer of plant resistance. Yeast cells provided maize plants with fewer rot symptoms and higher activities of enzymes related to plant resistance. Thus, we concluded that T. globosa (strain CCA6S01) might be used in agriculture practice as a plant protection agent. It can help to decrease the application of fungicides in the field and maintain plant productivity under stress.

Citrus essential oil: would it be feasible as antimicrobial in the bioethanol industry?

Essential oils (EOs) extracted from Citrus peels contain 85%-99% volatile components (a mixture of monoterpenes, sesquiterpenes, and their oxygenated derivatives) and 1%-15% non-volatile compounds. Citrus EOs have been long known for their antimicrobial properties, owing to which these EOs have a diverse range of applications. However, no studies have reported the applicability of Citrus EOs for the control of bacterial and yeast contaminants in the bioethanol industry. In this regard, the present review aimed to explore the feasibility of Citrus EOs in this industry. The Web of Science database was searched for reports that described the association of Citrus EOs with the most common microorganisms in the bioethanol industry to evaluate the efficacy of these EOs as antimicrobial agents in this context. The objective of the review was to suggest a novel antimicrobial that could replace sulfuric acid and antibiotics as the commonly used antimicrobial agents in the bioethanol industry. Citrus EOs exhibit antibacterial activity against Lactobacillus, which is the main bacterial genus that contaminates this fermentation process. The present report also confirms the selective action of these EOs on the contaminating yeasts and not/less on ethanol-producing yeast Saccharomyces cerevisiae, however further studies should be conducted to investigate the effects of Citrus EOs in yeast-bacterium co-culture.

Phosphate solubilization and indole acetic acid production by rhizosphere yeast Torulaspora globosa: improvement of culture conditions for better performance in vitro.

The rhizosphere yeast Torulaspora globosa is known to produce indole acetic acid (IAA) and to solubilize minerals. Due to the prospective use of this yeast as a biostimulant for agricultural applications, this work aimed to optimize the cultural conditions for both IAA production and phosphate solubilization. For phosphate solubilization, the temperature (20, 25 and 30 °C), initial medium pH (3.0, 5.0, and 7.0), and shaker speed (without mixing, 100 rpm, 150 rpm, and 200 rpm) were considered using the one-factor-at-a-time (OFAT) design. Temperature of 25 °C, initial medium pH 7.0, and static cultures were the conditions of greatest phosphate solubilization, with 40% of the total phosphorus content solubilized from calcium phosphate (419.86 mg L-1) after 48 h. By using the response surface methodology, the maximum IAA production (217.73 µg mL-1) was obtained with the highest initial pH 7.0, the lowest nitrogen, and glucose concentrations (5 g L-1 and 10 g L-1, respectively) and the lowest agitator speed (100 rpm). Further tests indicated that nitrogen affected significantly IAA production and the absence of nitrogen in the medium promoted higher IAA production (457 µg mL-1). The results obtained here may contribute to the scaling up for industrial and agricultural applications of a yeast-based product with T. globosa.

Saccharomyces cerevisiae strains used industrially for bioethanol production.

Fuel ethanol is produced by the yeast Saccharomyces cerevisiae mainly from corn starch in the United States and from sugarcane sucrose in Brazil, which together manufacture ∼85% of a global yearly production of 109.8 million m3 (in 2019). While in North America genetically engineered (GE) strains account for ∼80% of the ethanol produced, including strains that express amylases and are engineered to produce higher ethanol yields; in South America, mostly (>90%) non-GE strains are used in ethanol production, primarily as starters in non-aseptic fermentation systems with cell recycling. In spite of intensive research exploring lignocellulosic ethanol (or second generation ethanol), this option still accounts for <1% of global ethanol production. In this mini-review, we describe the main aspects of fuel ethanol production, emphasizing bioprocesses operating in North America and Brazil. We list and describe the main properties of several commercial yeast products (i.e., yeast strains) that are available worldwide to bioethanol producers, including GE strains with their respective genetic modifications. We also discuss recent studies that have started to shed light on the genes and traits that are important for the persistence and dominance of yeast strains in the non-aseptic process in Brazil. While Brazilian bioethanol yeast strains originated from a historical process of domestication for sugarcane fermentation, leading to a unique group with significant economic applications, in U.S.A., guided selection, breeding and genetic engineering approaches have driven the generation of new yeast products for the market.

Sequential process of solid-state cultivation with fungal consortium and ethanol fermentation by Saccharomyces cerevisiae from sugarcane bagasse.

Solid-state cultivation (SSC) is the microbial growth on solid supports, producing a nutrient-rich solution by cell enzymes that may be further used as a generic microbial medium. "Second-generation" ethanol is obtained by fermentation from mainly the acid hydrolysates of lignocellulosic wastes, generating several microbial growth inhibitors. Thus, this research aimed at evaluating the feasibility of ethanol fermentation from sugarcane bagasse hydrolysate after SSC with vinasse as the impregnating solution by a consortium of A. niger and T. reesei as opposed to the conventional method of acid hydrolysis. Fermentation of the hydrolysate from SSC leading to the yield of 0.40 g g-1, i.e., about 78% of maximum stoichiometric indicating that the nonconventional process allowed the use of two by-products from sugarcane processing in addition to ethanol production from glucose release.

From baker's yeast to genetically modified budding yeasts: the scientific evolution of bioethanol industry from sugarcane.

The peculiarities of Brazilian fuel ethanol fermentation allow the entry of native yeasts that may dominate over the starter strains of Saccharomyces cerevisiae and persist throughout the sugarcane harvest. The switch from the use of baker's yeast as starter to selected budding yeasts obtained by a selective pressure strategy was followed by a wealth of genomic information that enabled the understanding of the superiority of selected yeast strains. This review describes how the process of yeast selection evolved in the sugarcane-based bioethanol industry, the selection criteria and recent advances in genomics that could advance the fermentation process. The prospective use of genetically modified yeast strains, specially designed for increased robustness and product yield, with special emphasis on those obtained by the CRISPR (clustered regularly interspaced palindromic repeats)-Cas9 (CRISPR-associated protein 9) genome-editing approach, is discussed as a possible solution to confer higher performance and stability to the fermentation process for fuel ethanol production.

Agro-industrial wastes as sources of bioactive compounds for food and fermentation industries / Resíduos agroindustriais como fontes de compostos bioativos para as indústrias de alimentos e de fermentação

ABSTRACT: Agro-industrial wastes are of great interest because they are important natural sources of bioactive compounds and can generate value-added byproducts. Recent studies have shown that byproducts generated by the food processing industry are rich in bioactive compounds, such as phenolic compounds, organic acids, and carotenoids, among others. The growing interest in replacing synthetic antioxidant and antimicrobial compounds with natural ones has stimulated a search for these bioactive compounds from plant sources, especially from fruit-processing wastes. The study aims to project the agro-industrial wastes as potential natural sources of antioxidants and antimicrobials and the feasible technological applications in food and fermentation industries, especially the bioethanol industry.

RESUMO: Resíduos agroindustriais são de grande interesse por serem importantes fontes naturais de compostos bioativos e assim gerar subprodutos de valor agregado. Estudos recentes mostraram que o processamento de alimentos gera subprodutos ricos em compostos bioativos tais como compostos fenólicos, ácidos orgânicos e carotenóides, entre outros. O crescente interesse em substituir compostos antioxidantes e antimicrobianos sintéticos por naturais tem estimulado a procura por esses compostos bioativos a partir de fontes vegetais, especialmente de resíduos de processamento de frutas. O objetivo desta revisão é destacar os resíduos agroindustriais como potenciais fontes naturais de antioxidantes e antimicrobianos assim como as possíveis aplicações tecnológicas nas indústrias de alimentos e de fermentação, especialmente a indústria do bioetanol.

Volatile phenols are produced by strains of Dekkera bruxellensis under Brazilian fuel ethanol industry-like conditions.

Dekkera bruxellensis is a spoilage yeast in wine and fuel ethanol fermentations able to produce volatile phenols from hydroxycinnamic acids by the action of the enzymes cinnamate decarboxylase (CD) and vinyphenol reductase (VR) in wine. However, there is no information about this ability in the bioethanol industry. This work evaluated CD and VR activities and 4-ethylphenol production from p-coumaric acid by three strains of D. bruxellensis and PE-2, an industrial Saccharomyces cerevisiae strain. Single and multiple-cycle batch fermentations in molasses and sugarcane juice were carried out. Dekkera bruxellensis strains showed similar CD activity but differences in VR activity. No production of 4-ethylphenol by S. cerevisiae in any fermentation system or media was observed. The concentrations of 4-ethylphenol peaked during active growth of D. bruxellensis in single-cycle fermentation but they were lower than in multiple-cycle fermentation. Higher concentrations were observed in molasses with molar conversion (p-coumaric acid to 4-ethylphenol) ranging from 45% to 85%. As the first report on 4-ethylphenol production in sugarcane musts by D. bruxellensis in industry-like conditions, it opens up a new avenue to investigate its effect on the viability and fermentative capacity of S. cerevisiae as well as to understand the interaction between the yeasts in the bioethanol industry.

Conventional and nonconventional strategies for controlling bacterial contamination in fuel ethanol fermentations.

Ethanol bio-production in Brazil has some unique characteristics that inevitably lead to bacterial contamination, which results in the production of organic acids and biofilms and flocculation that impair the fermentation yield by affecting yeast viability and diverting sugars to metabolites other than ethanol. The ethanol-producing units commonly give an acid treatment to the cells after each fermentative cycle to decrease the bacterial number, which is not always effective. An alternative strategy must be employed to avoid bacterial multiplication but must be compatible with economic, health and environmental aspects. This review analyzes the issue of bacterial contamination in sugarcane-based fuel ethanol fermentation, and the potential strategies that may be utilized to control bacterial growth besides acid treatment and antibiotics. We have emphasized the efficiency and suitability of chemical products other than acids and those derived from natural sources in industrial conditions. In addition, we have also presented bacteriocins, bacteriophages, and beneficial bacteria as non-conventional antimicrobial agents to mitigate bacterial contamination in the bioethanol industry.

Interaction of Saccharomyces cerevisiae-Lactobacillus fermentum-Dekkera bruxellensis and feedstock on fuel ethanol fermentation.

The alcoholic fermentation for fuel ethanol production in Brazil occurs in the presence of several microorganisms present with the starter strain of Saccharomyces cerevisiae in sugarcane musts. It is expected that a multitude of microbial interactions may exist and impact on the fermentation yield. The yeast Dekkera bruxellensis and the bacterium Lactobacillus fermentum are important and frequent contaminants of industrial processes, although reports on the effects of both microorganisms simultaneously in ethanolic fermentation are scarce. The aim of this work was to determine the effects and interactions of both contaminants on the ethanolic fermentation carried out by the industrial yeast S. cerevisiae PE-2 in two different feedstocks (sugarcane juice and molasses) by running multiple batch fermentations with the starter yeast in pure or co-cultures with D. bruxellensis and/or L. fermentum. The fermentations contaminated with D. bruxellensis or L. fermentum or both together resulted in a lower average yield of ethanol, but it was higher in molasses than that of sugarcane juice. The decrease in the CFU number of S. cerevisiae was verified only in co-cultures with both D. bruxellensis and L. fermentum concomitant with higher residual sucrose concentration, lower glycerol and organic acid production in spite of a high reduction in the medium pH in both feedstocks. The growth of D. bruxellensis was stimulated in the presence of L. fermentum resulting in a more pronounced effect on the fermentation parameters than the effects of contamination by each microorganism individually.

Single and combined effects of acetic acid, furfural, and sugars on the growth of the pentose-fermenting yeast Meyerozyma guilliermondii.

The tolerance of the pentose-fermenting yeast Meyerozyma guilliermondii to the inhibitors released after the biomass hydrolysis, such as acetic acid and furfural, was surveyed. We first verified the effects of acetic acid and cell concentrations and initial pH on the growth of a M. guilliermondii strain in a semi-synthetic medium containing acetic acid as the sole carbon source. Second, the single and combined effects of furfural, acetic acid, and sugars (xylose, arabinose, and glucose) on the sugar uptake, cell growth, and ethanol production were also analysed. Growth inhibition occurred in concentrations higher than 10.5 g l-1 acetic acid and initial pH 3.5. The maximum specific growth rate (µ) was 0.023 h-1 and the saturation constant (ks) was 0.75 g l-1 acetic acid. Initial cell concentration also influenced µ. Acetic acid (initial concentration 5 g l-1) was co-consumed with sugars even in the presence of 20 mg l-1 furfural without inhibition to the yeast growth. The yeast grew and fermented sugars in a sugar-based medium with acetic acid and furfural in concentrations much higher than those usually found in hemicellulosic hydrolysates.

Bioethanol strains of Saccharomyces cerevisiae characterised by microsatellite and stress resistance

Abstract Strains of Saccharomyces cerevisiae may display characteristics that are typical of rough-type colonies, made up of cells clustered in pseudohyphal structures and comprised of daughter buds that do not separate from the mother cell post-mitosis. These strains are known to occur frequently in fermentation tanks with significant lower ethanol yield when compared to fermentations carried out by smooth strains of S. cerevisiae that are composed of dispersed cells. In an attempt to delineate genetic and phenotypic differences underlying the two phenotypes, this study analysed 10 microsatellite loci of 22 S. cerevisiae strains as well as stress resistance towards high concentrations of ethanol and glucose, low pH and cell sedimentation rates. The results obtained from the phenotypic tests by Principal-Component Analysis revealed that unlike the smooth colonies, the rough colonies of S. cerevisiae exhibit an enhanced resistance to stressful conditions resulting from the presence of excessive glucose and ethanol and high sedimentation rate. The microsatellite analysis was not successful to distinguish between the colony phenotypes as phenotypic assays. The relevant industrial strain PE-2 was observed in close genetic proximity to rough-colony although it does not display this colony morphology. A unique genetic pattern specific to a particular phenotype remains elusive.

Bioethanol strains of Saccharomyces cerevisiae characterised by microsatellite and stress resistance.

Strains of Saccharomyces cerevisiae may display characteristics that are typical of rough-type colonies, made up of cells clustered in pseudohyphal structures and comprised of daughter buds that do not separate from the mother cell post-mitosis. These strains are known to occur frequently in fermentation tanks with significant lower ethanol yield when compared to fermentations carried out by smooth strains of S. cerevisiae that are composed of dispersed cells. In an attempt to delineate genetic and phenotypic differences underlying the two phenotypes, this study analysed 10 microsatellite loci of 22 S. cerevisiae strains as well as stress resistance towards high concentrations of ethanol and glucose, low pH and cell sedimentation rates. The results obtained from the phenotypic tests by Principal-Component Analysis revealed that unlike the smooth colonies, the rough colonies of S. cerevisiae exhibit an enhanced resistance to stressful conditions resulting from the presence of excessive glucose and ethanol and high sedimentation rate. The microsatellite analysis was not successful to distinguish between the colony phenotypes as phenotypic assays. The relevant industrial strain PE-2 was observed in close genetic proximity to rough-colony although it does not display this colony morphology. A unique genetic pattern specific to a particular phenotype remains elusive.

Deterioration and fermentability of energy cane juice / Deterioração e fermentabilidade do caldo de cana energia

ABSTRACT: The main interest in the energy cane is the bioenergy production from the bagasse. The juice obtained after the cane milling may constitute a feedstock for the first-generation ethanol units; however, little attention has been dedicated to this issue. In order to verify the feasibility of the energy cane juice as substrate for ethanol production, the objectives of this research were first to determine the microbiological characteristics and deterioration along the time of the juices from two clones of energy cane (Type I) and second, their fermentability as feedstock for utilization in ethanol distilleries. There was a clear differentiation in the bacterial and yeast development of the sugarcane juices assayed, being much faster in the energy canes than in sugarcane. The storage of juice for 8 hours at 30oC did not cause impact in alcoholic fermentation for any sample analyzed, although a significant bacterial growth was detected in this period. A decrease of approximately seven percentage points in the fermentative efficiency was observed for energy cane juice in relation to sugarcane in a 24-hour fermentation cycle with the baking yeast. Despite the faster deterioration, the present research demonstrated that the energy cane juice has potential to be used as feedstock in ethanol-producing industries. As far as we know, it is the first research to deal with the characteristics of deterioration and fermentability of energy cane juices.

RESUMO: O principal interesse na cana energia reside na produção de bioenergia a partir do bagaço. O caldo obtido após a moagem da cana pode se constituir em substrato para as unidades de produção de etanol de primeira geração, no entanto, pouca atenção tem sido dispensada a esta questão. O presente trabalho avaliou o caldo de cana energia obtido de dois clones Tipo I como substrato para a produção de etanol, com base na determinação das suas características microbiológicas e deterioração ao longo do tempo, em comparação com o caldo de cana-de-açúcar (variedade RB867515). Foi observada uma clara diferenciação quanto ao crescimento bacteriano e de leveduras nas amostras de caldo analisadas, sendo o crescimento mais rápido no caldo de cana energia que no caldo de cana-de-açúcar. A manutenção do caldo por 8 horas a 30oC não causou impacto sobre a fermentação etanólica para quaisquer das amostras analisadas, apesar do crescimento significativo de bactérias. Houve um decréscimo de aproximadamente sete pontos percentuais na eficiência da fermentação com caldo de cana energia em um ciclo fermentativo de 24 horas com a levedura da panificação, em relação ao caldo da cana-de-açúcar. Apesar de a deterioração do caldo da cana energia ter sido mais rápida que a apresentada pelo caldo de cana-de-açúcar, o presente trabalho demonstrou que o caldo de cana energia tem potencial para ser utilizado como substrato nas indústrias produtoras de etanol. Do que se tem conhecimento, esse é o primeiro trabalho que trata das características de deterioração e fermentabilidade do caldo de cana energia.

A strain of Meyerozyma guilliermondii isolated from sugarcane juice is able to grow and ferment pentoses in synthetic and bagasse hydrolysate media.

The search for new microbial strains that are able to withstand inhibitors released from hemicellulosic hydrolysis and are also still able to convert sugars in ethanol/xylitol is highly desirable. A yeast strain isolated from sugarcane juice and identified as Meyerozyma guilliermondii was evaluated for the ability to grow and ferment pentoses in synthetic media and in sugarcane bagasse hydrolysate. The yeast grew in xylose, arabinose and glucose at the same rate at an initial medium pH of 5.5. At pH 4.5, the yeast grew more slowly in arabinose. There was no sugar exhaustion within 60 h. At higher xylose concentrations with a higher initial cell concentration, sugar was exhausted within 96 h at pH 4.5. An increase of 350 % in biomass was obtained in detoxified hydrolysates, whereas supplementation with 3 g/L yeast extract increased biomass production by approximately 40 %. Ethanol and xylitol were produced more significantly in supplemented hydrolysates regardless of detoxification. Xylose consumption was enhanced in supplemented hydrolysates and arabinose was consumed only when xylose and glucose were no longer available. Supplementation had a greater impact on ethanol yield and productivity than detoxification; however, the product yields obtained in the present study are still much lower when compared to other yeast species in bagasse hydrolysate. By the other hand, the fermentation of both xylose and arabinose and capability of withstanding inhibitors are important characteristics of the strain assayed.

Industrial PE-2 strain of Saccharomyces cerevisiae: from alcoholic fermentation to the production of recombinant proteins.

Saccharomyces cerevisiae is the most important microorganism used in the ethanol fermentation process. The PE-2 strain of this yeast is widely used to produce alcohol in Brazil due to its high fermentation capacity. The aim of the present study was to develop an expression system for recombinant proteins using the industrial PE-2 strain of S. cerevisiae during the alcoholic fermentation process. The protein chosen as a model for this system was CaneCPI-1, a cysteine peptidase inhibitor. A plasmid containing the CaneCPI-1 gene was constructed and yeast cells were transformed with the pYADE4_CaneCPI-1 construct. To evaluate the effect on fermentation ability, the transformed strain was used in the fermentation process with cell recycling. During the nine-hour fermentative cycles the transformed strain did not have its viability and fermentation ability affected. In the last cycle, when the fermentation lasted longer, the protein was expressed probably at the expense of ethanol once the sugars were exhausted. The recombinant protein was expressed in yeast cells, purified and submitted to assays of activity that demonstrated its functionality. Thus, the industrial PE-2 strain of S. cerevisiae can be used as a viable system for protein expression and to produce alcohol simultaneously. The findings of the present study demonstrate the possibility of producing recombinant proteins with biotechnological applications during the ethanol fermentation process.

Characteristics of Saccharomyces cerevisiae yeasts exhibiting rough colonies and pseudohyphal morphology with respect to alcoholic fermentation

Among the native yeasts found in alcoholic fermentation, rough colonies associated with pseudohyphal morphology belonging to the species Saccharomyces cerevisiae are very common and undesirable during the process. The aim of this work was to perform morphological and physiological characterisations of S. cerevisiae strains that exhibited rough and smooth colonies in an attempt to identify alternatives that could contribute to the management of rough colony yeasts in alcoholic fermentation. Characterisation tests for invasiveness in Agar medium, killer activity, flocculation and fermentative capacity were performed on 22 strains (11 rough and 11 smooth colonies). The effects of acid treatment at different pH values on the growth of two strains ("52" -rough and "PE-02" smooth) as well as batch fermentation tests with cell recycling and acid treatment of the cells were also evaluated. Invasiveness in YPD Agar medium occurred at low frequency; ten of eleven rough yeasts exhibited flocculation; none of the strains showed killer activity; and the rough strains presented lower and slower fermentative capacities compared to the smooth strains in a 48-h cycle in a batch system with sugar cane juice. The growth of the rough strain was severely affected by the acid treatment at pH values of 1.0 and 1.5; however, the growth of the smooth strain was not affected. The fermentative efficiency in mixed fermentation (smooth and rough strains in the same cell mass proportion) did not differ from the efficiency obtained with the smooth strain alone, most likely because the acid treatment was conducted at pH 1.5 in a batch cell-recycle test. A fermentative efficiency as low as 60% was observed with the rough colony alone.

Fermentative and growth performances of Dekkera bruxellensis in different batch systems and the effect of initial low cell counts in co-cultures with Saccharomyces cerevisiae.

Dekkera bruxellensis is a multifaceted yeast present in the fermentative processes used for alcoholic beverage and fuel alcohol production - in the latter, normally regarded as a contaminant. We evaluated the fermentation and growth performance of a strain isolated from water in an alcohol-producing unit, in batch systems with/without cell recycling in pure and co-cultures with Saccharomyces cerevisiae. The ethanol resistance and aeration dependence for ethanol/acid production were verified. Ethanol had an effect on the growth of D. bruxellensis in that it lowered or inhibited growth depending on the concentration. Acid production was verified in agitated cultures either with glucose or sucrose, but more ethanol was produced with glucose in agitated cultures. Regardless of the batch system, low sugar consumption and alcohol production and expressive growth were found with D. bruxellensis. Despite a similar ethanol yield compared to S. cerevisiae in the batch system without cell recycling, ethanol productivity was approximately four times lower. However, with cell recycling, ethanol yield was almost half that of S. cerevisiae. At initial low cell counts of D. bruxellensis (10 and 1000 cells/ml) in co-cultures with S. cerevisiae, a decrease in fermentative efficiency and a substantial growth throughout the fermentative cycles were displayed by D. bruxellensis. Due to the peculiarity of cell repitching in Brazilian fermentation processes, D. bruxellensis is able to establish itself in the process, even when present in low numbers initially, substantially impairing bioethanol production due to the low ethanol productivity, in spite of comparable ethanol yields.

Effects of single and combined cell treatments based on low pH and high concentrations of ethanol on the growth and fermentation of Dekkera bruxellensis and Saccharomyces cerevisiae.

The alcoholic fermentation in Brazil displays some peculiarities because the yeast used is recycled in a non-aseptic process. After centrifugation, the cells are treated with acid to control the bacterial growth. However, it is difficult to manage the indigenous yeasts without affecting the main culture of Saccharomyces cerevisiae. This work evaluated how the cell treatment could be modified to combat contaminant yeasts based on the differential sensitivities to low pH and high concentrations of ethanol displayed by an industrial strain of S. cerevisiae and three strains of Dekkera bruxellensis, which are common contaminant yeasts in Brazilian fermentation processes. The tests were initially performed in rich medium with a low pH or a high concentration of ethanol to analyse the yeast growth profile. Then, the single and combined effects of low pH and ethanol concentration on the yeast cell viability were evaluated under non-proliferative conditions. The effects on the fermentation parameters were also verified. S. cerevisiae grew best when not subjected to the stresses, but this yeast and D. bruxellensis had similar growth kinetics when exposed to a low pH or increased ethanol concentrations. However, the combined treatments of low pH (2.0) and ethanol (11 or 13 %) resulted in a decrease of D. bruxellensis cell viability almost three times higher than of S. cerevisiae, which was only slightly affected by all cell treatments. The initial viability of the treated cells was restored within 8 h of growth in sugar cane juice, with the exception of the combined treatment for D. bruxellensis. The ethanol-based cell treatment, in despite of slowing the fermentation, could decrease and maintain D. bruxellensis population under control while S. cerevisiae was taking over the fermentation along six fermentative cycles. These results indicate that it may be possible to control the growth of D. bruxellensis without major effects on S. cerevisiae. The cells could be treated between the fermentation cycles by the parcelled addition of 13 % ethanol to the tanks in which the yeast cream is treated with sulphuric acid at pH 2.0.