Enhanced production of N-acetyl-glucosaminidase by marine Aeromonas caviae CHZ306 in bioreactor.

N-Acetyl-glucosaminidases (GlcNAcases) are exoenzymes found in a wide range of living organisms, which have gained great attention in the treatment of disorders related to diabetes, Alzheimer's, Tay-Sachs', and Sandhoff's diseases; the control of phytopathogens; and the synthesis of bioactive GlcNAc-containing products. Aiming at future industrial applications, in this study, GlcNAcase production by marine Aeromonas caviae CHZ306 was enhanced first in shake flasks in terms of medium composition and then in bench-scale stirred-tank bioreactor in terms of physicochemical conditions. Stoichiometric balance between the bioavailability of carbon and nitrogen in the formulated culture medium, as well as the use of additional carbon and nitrogen sources, played a central role in improving the bioprocess, considerably increasing the enzyme productivity. The optimal cultivation medium was composed of colloidal α-chitin, corn steep liquor, peptone A, and mineral salts, in a 5.2 C:N ratio. Optimization of pH, temperature, colloidal α-chitin concentration, and kLa conditions further increased GlcNAcase productivity. Under optimized conditions in bioreactor (i.e., 34 °C, pH 8 and kLa 55.2 h-1), GlcNAcase activity achieved 173.4 U.L-1 after 12 h of cultivation, and productivity no less than 14.45 U.L-1.h-1 corresponding to a 370-fold enhancement compared to basal conditions.

Bioproduction of α-terpineol and R-(+)-limonene derivatives by terpene-tolerant ascomycete fungus as a potential contribution to the citrus value chain.

AIMS: The aims of this article were to select fungal species with high tolerance and high growth rate in mediums supplemented with limonene and citrus essential oils (CEOs), and to test the bioconversion capability of the chosen isolates for the bioproduction of aroma compounds. METHODS AND RESULTS: Based on the use of predictive mycology, 21 of 29 isolates were selected after assaying R-(+)-limonene and CEO tolerance (10 g l-1 ). With a dendrogram divisive coefficient of 0·937, the subcluster two with isolates Aspergillus niger LBM 055, Penicillium sp. LBM 150, Penicillium sp. LBM 151 and Penicillium sp. LBM 154 gathered the highest tolerance and mycelia growth speed. Ultrastructural analysis indicated that culture media containing limonene had no visible toxic activity that could promote morphological changes in the fungal cell wall. The biomass of A. niger LBM055 was distinctive in liquid media supplemented with R-(+)-limonene (0·57 ± 0·07 g) and it was selected to prove bioconversion capacity, under static and agitated conditions, and converted up to 98% of limonene, yielding a wide variety of products that were quantified by GC-FID. It was obtained at molecular weights less than limonene (64-100%), between limonene and α-terpineol (12-72%) and greater than α-terpineol (2-48%). CONCLUSIONS: Aspergillus niger LBM 055, Penicillium sp. LBM 150, Penicillium sp. LBM 151 and Penicillium sp. LBM 154 showed to the highest tolerance and growth rate in mediums supplemented with R-(+)-limonene and orange and lemon essential oils. Particularly, A. niger LBM055, showed limonene bioconversion capability and produced different molecular weights compounds such us α-terpineol. SIGNIFICANCE AND IMPACT OF THE STUDY: Different bioproducts can be obtained by changing operative condition with the same fungus, and this bioprocess aspect is a significant approach to be adopted on industrial scale leading to the creation of new natural flavours and fragrance compositions.

Optimized Production of a Redox Metabolite (pyocyanin) by Pseudomonas aeruginosa NEJ01R Using a Maize By-Product.

Pseudomonas aeruginosa metabolizes pyocyanin, a redox molecule related to diverse biological activities. Culture conditions for the production of pyocyanin in a defined medium were optimized using a statistical design and response surface methodology. The obtained conditions were replicated using as substrate an alkaline residual liquid of cooked maize and its by-products. The untreated effluent (raw nejayote, RN) was processed to obtain a fraction without insoluble solids (clarified fraction, CL), then separated by a 30 kDa membrane where two fractions, namely, retentate (RE) and filtered (FI), were obtained. Optimal conditions in the defined medium were 29.6 °C, 223.7 rpm and pH = 6.92, which produced 2.21 µg mL-1 of pyocyanin, and by using the wastewater, it was possible to obtain 3.25 µg mL-1 of pyocyanin in the retentate fraction at 40 h. The retentate fraction presented the highest concentration of total solids related to the maximum concentration of pyocyanin (PYO) obtained. The pyocyanin redox behavior was analyzed using electrochemical techniques. In this way, valorization of lime-cooked maize wastewater (nejayote) used as a substrate was demonstrated in the production of a value-added compound, such as pyocyanin, a redox metabolite of Pseudomonas aeruginosa NEJ01R.

Production of volatile compounds by yeasts using hydrolysed grape seed oil obtained by immobilized lipases in continuous packed-bed reactors.

Lipases CAL-B, TLL, and RML were used in the synthesis of free fatty acids of grape seed oil as heterogeneous substrate. The best enzyme was used to optimize the reaction variables temperature, enzyme content, and molar ratio of water:oil in batch reactions using experimental planning. The ideal conditions to produce free fatty acids using pure RML were 45 °C, 12:1 substrate molar ratio, and 15% enzyme, resulting in 66% of oil hydrolysis and a productivity of 0.54 mol L-1 min-1 in 4 h of reaction at 180 rpm. Repeated batches of reaction were performed testing the operational stability of RML, results showing that this enzyme could be used for at least 20 cycles keeping more than 80% of its initial activity, suggesting its potential use in industrial processes. The synthesis of free fatty acids was then evaluated in continuous reactions using packed-bed reactor (PBR). The highest productivity in the continuous process was 6.85 mol L-1 min-1, using only RML, showing an operational stability higher than 80% of its initial conversion capacity after 11 days of operation, at a flow rate of 0.13 mL min-1 at 45 °C. We evaluated the use of this hydrolyzed oil as substrate for lactone bioproduction using Galactomyces geotrichum UFMG-CM-Y3276, G. geotrichum UFMG-CM-Y3558, and Geotrichum klebahnii UFMG-CM-Y3014 screened for their oil-hydrolysis ability. Volatile compounds were qualitatively identified in GC-MS as γ-octalactone and γ-nonalactone.

Influencia del hidrolizado de proteínas en el comportamiento bioproductivo en gallinas de la línea L1 White Leghorn / Influence of protein hydrolysates on bioproduction performance in L1 line White Leghorn hens

RESUMEN El objetivo de la presente investigación fue evaluar la influencia del hidrolizado de proteínas (HP) como suplemento nutricional en el comportamiento bioproductivo en gallinas White Leghorn. Se utilizaron gallinas de reproductores ligeros con 39 semanas de edad, durante seis semanas. Se realizó un diseño completamente aleatorizado con dos tratamientos de 320 gallinas cada uno (tres réplicas de 40 gallinas). El T1, recibió diariamente 2 mL de HP por ave y al T2, no se le ofertó el producto. Se controlaron las variables bioproductivas (producción total de huevos, porcentaje de postura, consumo total de pienso, conversión, viabilidad y la mortalidad por causas), los indicadores de la incubación (huevos a planta, los porcentajes de incubación, incubabilidad y pollitos de primera) y los de calidad (consumo de pienso por pollito de primera y pollitos de primera por gallina). Se observaron diferencias significativas en el porcentaje de postura y significativa para la conversión masal y en los huevos a planta, porcentaje de incubación y porcentaje de incubabilidad y con mejor comportamiento en los indicadores de calidad en las gallinas que recibieron HP. Se concluye que el empleo de HP mejora el porcentaje de postura y la conversión masal y los indicadores relativos a la incubación, porcentaje de huevos aptos a planta, porcentaje de incubación y de incubabilidad de gallinas de la línea ligera, adicionalmente, su uso reduce aproximadamente en 100 gramos el consumo de pienso necesario para obtener un pollito de primera.

ABSTRACT The objective of this study was to evaluate the influence of protein hydrolysates (PH), as a nutritional supplement, in the bioproduction performance of White Leghorn hens. Thirty-nine-week old light line hens were assessed during six weeks. Two treatments (T) were designed involving 320 hens each (three replications of 40 hens each). In T1, each bird received 2 ml of PH daily; the birds in T2 were not offered the product. The following variables were controlled: bioproduction (total egg production, egg-laying percentage, total feed intake, mass conversion, viability, and caused mortality); incubation indicators (eggs to plant, incubation percentage, hatchability, and top quality chicks) and; quality (feed intake by top quality chicks, and top quality chicks per hen). The hens that received the PH showed significant differences in the laying, incubation, and hatchability percentage, as well as mass conversion, and the egg to plant ratio. They showed better performance in quality indicators. It is concluded that the use of PH in light line hens improves their bioproduction performance, as well as the indicators of to incubation, egg to plant ratio, and hatchability; its use also reduces by approximately 100 grams the necessary food intake to obtain top quality chicks.

An overview of biotechnological production of propionic acid: from upstream to downstream processes

The increasing demand for propionic acid (PA) production and its wide applications in several industries, especially the food industry (as a preservative and satiety inducer), have led to studies on the low-cost biosynthesis of this acid. This paper gives an overview of the biotechnological aspects of PA production and introduces Propionibacterium as the most popular organism for PA production. Moreover, all process variables influencing the production yield, different simple and complex carbon sources, the metabolic pathway of production, engineered mutants with increased productivity, and modified tolerance against high concentrations of acid have been described. Furthermore, possible methods of extraction and analysis of this organic acid, several applied bioreactors, and different culture systems and substrates are introduced. It can be concluded that maximum biomass and PA production may be achieved using metabolically engineered microorganisms and analyzing the most significant factors influencing yield. To date, the maximum reported yield for PA production is 0.973 g·g-1, obtained from Propionibacterium acidipropionici in a three-electrode amperometric culture system in medium containing 0.4 mM cobalt sepulchrate. In addition, the best promising substrate for PA bioproduction may be achieved using glycerol as a carbon source in an extractive continuous fermentation. Simultaneous production of PA and vitamin B12 is suggested, and finally, the limitations of and strategies for competitive microbial production with respect to chemical process from an economical point of view are proposed and presented. Finally, some future trends for bioproduction of PA are suggested.