Selenium-Enriched Probiotic Saccharomyces boulardii CCT 4308 Biomass Production Using Low-Cost Sugarcane Molasses Medium

Abstract Food supplements have been increasingly investigated. Probiotics have several benefits for human and animal health and selenium (Se) is widely recommended against oxidative stress. In this context, the aim of this study was to develop a low-cost bioprocess to produce a functional food product comprising both probiotic and Se accumulation. Yeast cells of Saccharomyces boulardii CCT 4308 were cultivated using sugarcane molasses as substrate. Optimization studies were performed to evaluate the best medium composition for biomass production and Se-accumulation in batch and fed-batch systems. Optimized conditions were defined with a medium composed of 150 g L-1 sugarcane molasses and 12 g L-1 yeast extract, with feeding of 100 g L-1 sugarcane molasses and 100 μg mL-1 of Se incorporation after 4 h and 10 h of fermentation, respectively, during 48 h in STR (stirred tank reactor). Best biomass production reached 14.52 g L-1 with 3.20 mg Se g-1 biomass at 12 h. Process optimization led to 4.82-fold increase in biomass production compared to initial condition. A final Se-enriched S. boulardii CCT 4308 biomass was obtained, which is comparable to commercial products. An alternative probiotic yeast biomass was efficiently produced as a new food-form of Se supplement in a sustainable process using an inexpensive agro-industrial residue.

Comparison of photoautotrophic and mixotrophic cultivation of microalgae Messastrum gracile (Chlorophyceae) in alternative culture media / Comparação dos cultivos foto-autotrófico e mixotrófico da microalga Messastrum gracile (Chlorophyceae) em meios de cultura alternativos

Abstract Growth and biological conditions of Messastrum gracile were evaluated to compare the effect of photoautotrophic and mixotrophic cultivation on the increase of biomass production and chemical conditions cultured in macrophyte and commercial culture media. The growth rate (k) of M. gracile was different in the culture media, higher in mixotrophic cultivation for Lemna minor culture medium, whilst to Eichhornia crassipes and NPK culture media were higher in photoautotrophic cultivation. Mean lipid contents in photoautotrophic cultivation were 8.2% biomass dry weight, whereas they reached 19% biomass dry weight in mixotrophic cultivation. Protein contents were below 48% biomass dry weight in photoautotrophic cultivation and 30% biomass dry weight in mixotrophic cultivation. Messastrum gracile cultured in macrophyte culture media (E. crassipes and L. minor) and NPK culture medium provided satisfactory results with regard to lipid and protein contents in mixotrophic and photoautotrophic cultivations, respectively. Lipid and protein contents in alternative media were higher or similar to the CHU12 commercial culture medium.

Resumo O crescimento e as condições biológicas da microalga Messastrum gracile foram avaliados para comparar o efeito do cultivo foto-autotrófico e mixotrófico na produção de biomassa e condições químicas em meios de cultura comercial e de macrófitas. A taxa de crescimento (k) de M. gracile foi diferente entre os meios de cultura, sendo maior no cultivo mixotrófico para o meio Lemna minor, enquanto para os meios Eichhornia crassipes e NPK foram maiores no cultivo foto-autotrófico. Os teores de lipídios no cultivo foto-autotrófico foram de 8,2% da biomassa seca, enquanto que no mixotrófico atingiram 19% da biomassa seca. Os teores de proteína em cultivo foto-autotrófico estiveram abaixo de 48% da biomassa seca e 30% de biomassa seca no cultivo mixotrófico. Messastrum gracile cultivada em meios de cultura de macrófitas (E. crassipes e L. minor) e NPK apresentaram resultados satisfatórios em relação aos teores de lipídeos e proteínas nos cultivos mixotróficos e foto-autotróficos, respectivamente. Os teores lipídicos e proteicos em meios alternativos foram maiores ou semelhantes ao meio comercial CHU12.

Bioconversion of sugarcane molasses to poly(3-hydroxybutyrate-co-3- hydroxyvalerate) by endophytic Bacillus cereus RCL 02

Sugarcane molasses, the by-product of sugar industry are rich in nutrients, growth factors and minerals and areefficiently utilized by the microorganisms for growth and production of primary as well as secondary metabolitesof commercial importance. In this communication, we report the utilization of sugarcane molasses as the solesource of carbon for the production of copolymers of polyhydroxyalkanoates (PHAs) with improved materialproperties. The endophytic bacterium Bacillus cereus RCL 02 (MCC 3436) produced 7.8 g/L of PHA whengrown in MS medium with 4% (w/v) sugarcane molasses under batch cultivation. Proton nuclear magneticresonance (1H NMR) analysis revealed that the copolymer so produced contain 12.4 mol% 3-hydroxyvalerate(3HV) along with 87.6 mol% 3-hydroxybutyrate (3HB). The copolyester, P(3HB-co-12.4 mol%-3HV)has been isolated and purified following standard solvent extraction method and partially characterized byFourier-transform infrared (FTIR), X-ray diffraction (XRD), thermogravimetric (TG) and differential scanningcalorimetric (DSC) analysis. The material and thermal properties of the copolyester so produced indicated itspotential for industrial application.

Comparison of photoautotrophic and mixotrophic cultivation of microalgae Messastrum gracile (Chlorophyceae) in alternative culture media

Abstract Growth and biological conditions of Messastrum gracile were evaluated to compare the effect of photoautotrophic and mixotrophic cultivation on the increase of biomass production and chemical conditions cultured in macrophyte and commercial culture media. The growth rate (k) of M. gracile was different in the culture media, higher in mixotrophic cultivation for Lemna minor culture medium, whilst to Eichhornia crassipes and NPK culture media were higher in photoautotrophic cultivation. Mean lipid contents in photoautotrophic cultivation were 8.2% biomass dry weight, whereas they reached 19% biomass dry weight in mixotrophic cultivation. Protein contents were below 48% biomass dry weight in photoautotrophic cultivation and 30% biomass dry weight in mixotrophic cultivation. Messastrum gracile cultured in macrophyte culture media (E. crassipes and L. minor) and NPK culture medium provided satisfactory results with regard to lipid and protein contents in mixotrophic and photoautotrophic cultivations, respectively. Lipid and protein contents in alternative media were higher or similar to the CHU12 commercial culture medium.

Resumo O crescimento e as condições biológicas da microalga Messastrum gracile foram avaliados para comparar o efeito do cultivo foto-autotrófico e mixotrófico na produção de biomassa e condições químicas em meios de cultura comercial e de macrófitas. A taxa de crescimento (k) de M. gracile foi diferente entre os meios de cultura, sendo maior no cultivo mixotrófico para o meio Lemna minor, enquanto para os meios Eichhornia crassipes e NPK foram maiores no cultivo foto-autotrófico. Os teores de lipídios no cultivo foto-autotrófico foram de 8,2% da biomassa seca, enquanto que no mixotrófico atingiram 19% da biomassa seca. Os teores de proteína em cultivo foto-autotrófico estiveram abaixo de 48% da biomassa seca e 30% de biomassa seca no cultivo mixotrófico. Messastrum gracile cultivada em meios de cultura de macrófitas (E. crassipes e L. minor) e NPK apresentaram resultados satisfatórios em relação aos teores de lipídeos e proteínas nos cultivos mixotróficos e foto-autotróficos, respectivamente. Os teores lipídicos e proteicos em meios alternativos foram maiores ou semelhantes ao meio comercial CHU12.

Optimization of lipids production by Cryptococcus laurentii 11 using cheese whey with molasses

This study aimed the optimization of culture condition and composition for production of Cryptococcus laurentii 11 biomass and lipids in cheese whey medium supplemented with sugarcane molasses. The optimization of pH, fermentation time, and molasses concentration according to a full factorial statistical experimental design was followed by a Plackett-Burman experimental design, which was used to determine whether the supplementation of the culture medium by yeast extract and inorganic salts could provide a further enhancement of lipids production. The following conditions and composition of the culture medium were found to optimize biomass and lipids production: 360 h fermentation, 6.5 pH and supplementation of (g L-1): 50 molasses, 0.5 yeast extract, 4 KH2PO4, 1 Na2HPO4, 0.75 MgSO4•7H2O and 0.002 ZnSO4•H2O. Additional supplementation with inorganic salts and yeast extract was essential to optimize the production, in terms of product concentration and productivity, of neutral lipids by C. laurentii 11. Under this optimized condition, the production of total lipids increased by 133% in relation to control experiment (from 1.27 to 2.96 g L-1). The total lipids indicated a predominant (86%) presence of neutral lipids with high content of 16- and 18- carbon-chain saturated and monosaturated fatty acids. This class of lipids is considered especially suitable for the production of biodiesel.

Optimization of biomass production with copper bioaccumulation by yeasts in submerged fermentation

The objective of this work was to study the production of biomass with copper bioaccumulation in submerged fermentation using sugarcane molasses. Candida pelliculosa BARU 05 isolated from Baru (Dipteryx alata) was selected for its good capacity to accumulate the copper. Fermentation was carried out using the medium composed by sugarcane molasses at 5 °Brix enriched with (g/L) CuSO4.5H2O 0.1; yeast extract, 10.0; (NH4)2SO4, 5.0 ; KH2PO4, 5.0 MgSO4, 0.5, inoculum 10 percent of total volume (100 ml), pH 6.0, and incubation at 30 °C, 120 rpm for 120 h. After three steps of optimization an uptake of 95.04 percent and 13.397 g/L biomass were obtained. The kinetics of copper bioaccumulation and biomass production was followed in a 10- liter bioreactor in a batch and fed-batch fermentation which showed copper accumulation of 91.98 and 100 percent, respectively, and biomass production of 38.85 g/L (24 h) and 57.54 g/L (48 h), respectively.

Development of a Low Cost Bioprocess for Endotoxin Production by Bacillus thuringiensis var israelensis Intended for Biological Control of Aedes aegypti

Aedes aegypti is the vector of Dengue disease, responsible for 20,000 deaths/year worldwide. Bacillus thuringiensis var israelensis - Bti releases selective and effective toxins (crystal proteins) against A. aegypti larvae. We present a low cost bioprocess for toxin production, accomplished by a selected Brazilian strain Bti (BR-LPB01) and employment of low cost substrates. Soybean meal and sugarcane molasses lead to high toxic effectiveness after 2L bioreactor fermentation (LD50=26ng/mL), near to the reference strain IPS82 (LD50=17.3 ng/mL). The pH ranged between 5.8 and 7.0 during the exponential growth period and between 7.0 and 8.4 during the stationary phase, with low activity. Thus, control of foam and pH 7.0 were started and proved to be crucial for high activity. It was verified that the fermentation could be discontinued after 20 hours, when the highest activity was present.

A dengue é transmitida pelo Aedes aegypti, doença responsável por 20.000 mortes/ano no mundo. Bacillus thuringiensis var israelensis libera toxinas seletivas e eficazes (proteínas cristal) contra larvas de A. aegypti. Propõe-se um bioprocesso de baixo custo para a produção da toxina, pelo emprego de uma cepa brasileira selecionada de Bti (BR-LPB01) e de substratos de baixo custo. Farelo de soja e melaço de cana levaram a eficácia tóxica alta após fermentação em biorreator 2L (DL50=26ng/mL), valor próximo a estirpe de referência IPS82 (DL50=17,3 ng/mL). O pH variou entre 5,8 e 7,0 durante o período de crescimento exponencial e entre 7,0 e 8,4 durante a fase estacionária, com baixa atividade larvicida. Assim, controles de espuma e de pH 7,0 foram iniciados e demonstraram serem cruciais para alta atividade. Verificou-se que a fermentação deve ser interrompida após vinte horas, quando se obtém a maior atividade.

Evaluación de la capacidad probiótica in vitro de una cepa nativa de Saccharomyces cerevisiae / In vitro evaluation of the probiotic capacity of a native strain of saccharomyces cerevisiae

En el presente estudio se evaluó la capacidad probiótica in vitro de una cepa nativa de Saccharomyces cerevisiae (A) y se comparó con una cepa comercial (B) utilizada como probiótico. Para esto se determinó la concentración de melaza de caña (10, 20 y 30 por ciento (p/v)) que permitiera obtener la mayor cantidad de biomasa de las cepas, así mismo se determinaron parámetros cinéticos. La concentración de melaza que arrojó mejores resultados fue 20por ciento (p/v) y se encontró diferencia en la producción de biomasa para la cepa en estudio A (28g/L) y la cepa control B (3g/L) en medio melaza. Se realizaron pruebas in vitro como resistencia a sales biliares, tolerancia a rangos de pH y jugos gástricos, donde no se observaron diferencias entre la cepa A y B al medir el crecimiento. La reducción del colesterol en presencia de sales biliares después de 12 horas de incubación fue de 54 por ciento para la cepa A y 58 por ciento para la B. Por último se realizó una prueba de adherencia en células Caco-2, encontrando adherencia a estas por parte de ambas cepas. De acuerdo con los resultados anteriores se demostró que la cepa A tiene propiedades probióticas in vitro que pueden ser corroboradas con posteriores estudios in vivo que confirmen su utilización como probiótico en nutrición animal.

The in vitro probiotic capacity of a native strain of Saccharomyces cerevisiae (A) was evaluated and compared with a commercial strain (B) used as a probiotic. The effect of the concentration of sugarcane molasses (10, 20 and 30% (w/v)) on the biomass production was investigated and kinetic parameters were determined. The best molasses concentration was 20% (w/v) and differences in biomass production on molasses medium between strain A (28 g/L) and control strain B (3 g/L) were observed. In vitro tests such as tolerance to bile salts, pH and gastric juices were carried out, and no differences in growth between strain A and B were found. Cholesterolreduction on presence of bile salts after 12 hours of incubation was of 54% for strain A and 58% for strain B. Both strains showed adherence to Caco-2 cells. Results reveal that strain A possesses in vitro probiotic propertiesthat can be verified with further in vivo studies to confirm its suitability as probiotic in animal nutrition.