Inhibition of Aspergillus carbonarius growth and Ochratoxin A production using lactic acid bacteria cultivated in an optimized medium.

The Gram-positive bacteria lactic acid bacteria (LAB) are used in the food industry but are also known for inhibiting certain food spoilage microorganisms, especially fungi. Sources of nitrogen (N) for culture media are generally organic and expensive. Many attempts have been made to formulate economical culture media with alternative N sources obtained from agricultural and industrial byproducts. This study describes the design and optimization of an inexpensive culture medium for Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) MZ809351 strain B31. The culture medium was optimized using statistical experimental designs to identify the factors with the most significant effects on biomass concentration to reduce the overall cost, aiming to obtain a biomass concentration similar to that obtained with the reference LAB culture medium (de Man, Rogosa and Sharpe; MRS). Sodium acetate and magnesium sulfate were the most significant factors (p < 0.005), and their contents were reduced by 22 % and 40 %, respectively, without affecting biomass concentration. Malt germ extract (MGE) was used as an alternative nitrogen source to replace meat extract (ME) and proteose peptone (PP). Through these experiments, the composition of a culture medium that is less expensive than MRS broth was defined, which produced a biomass concentration (3.8 g/L) similar to that obtained with MRS medium. The inhibitory effects of two LAB strains isolated from the Ivory Coast and Mexico on the growth and production of ochratoxin A (OTA) in an ochratoxigenic fungus was tested. The minimum cellular concentration of the LAB to prevent the development of Aspergillus carbonarius Ac 089 and the production of OTA was determined in a model assay in Petri dishes. The conditions to inhibit the germination of A. carbonarius Ac 089 and the production of OTA were found. Using the optimized medium and a ratio of 2 × 104 LAB/spore (1 × 108 CFU/mL) strain B7 (L. plantarum MZ809351) and 2 × 103 LAB/spore (1 × 107 CFU/mL) strain B31 (L. plantarum MN922335) completely inhibited the growth of the fungus. A ratio of 2 × 105 LAB/spore (1 × 109 CFU/mL) was required to inhibit OTA production with strains B7 and B31. This study indicates the potential of cultivating LAB in an optimized and inexpensive culture medium for use as a biological control agent against ochratoxigenic fungi in food.

Solid-state culture of Azospirillum brasilense: a reliable technology for biofertilizer production from laboratory to pilot scale.

A biofertilizer of Azospirillum brasilense was produced in solid-state culture (SSC) from laboratory to pilot scale. Similar operation conditions (continuous aeration and mild intermittent mixing) and two dimensionless numbers with similar L/D ratio and a similar working volume were applied to reach a scale-up factor of 75. An innovative bioreactor with rotating helical ribbons (15 kg wet matter) was used at pilot scale. A mathematical model was proposed and validated to evaluate the respirometry trends at laboratory and pilot scale exhibiting similar behavior. The cell viability was (1.3 ± 0.4) × 109 and (1.3 ± 0.3) × 109 colony-forming units per gram of initial dry mass at laboratory and pilot scale, at 36 and 43 h, respectively. A. brasilense maintains its viability twelve months of storage at 4 and 30 °C. This is the first report of A. brasilense being cultivated in SSC under controlled conditions. SSC processes involving unicellular microorganisms with tolerance to agitation are a promising technology to produce biofertilizers.

Effect of stirring on growth and cellulolytic enzymes production by Trichoderma harzianum in a novel bench-scale solid-state fermentation bioreactor.

A novel bench-scale stirred bioreactor for solid-state fermentation was used to determine the effect of the stirring rate on growth and enzymes production by Trichoderma harzianum PBLA. Lab-scale static tubular bioreactors were first used to assess the effect of bioreactor diameter on heat accumulation, growth, and production of cellulases and xylanases. The increased diameters (1.8-4.2 cm) led to increases in temperature up to 36 °C (at a rate of 1.08 °C/cm), which negatively affected the growth and enzyme production. Afterward, in the bench-scale bioreactor operated at rates up to 3.0 rpm, maximum xylanases production (107 ±â€¯0.3 U/g dm) was attained at rates of 0.5 and 1.0 rpm, reaching a maximum of 34 ±â€¯0.3 °C. Cellulases production was reduced (up to 79%) due to stirring. Therefore, the production of xylanases by T. harzianum can be performed in this cross-flow stirred SSF bioreactor at rates up to 1.0 rpm, avoiding heat accumulation and damage on metabolic activity.

Review on technological and scientific aspects of feruloyl esterases: A versatile enzyme for biorefining of biomass.

With increasing focus on sustainable energy, bio-refining from lignocellulosic biomass has become a thrust area of research. With most of the works being focused on biofuels, significant efforts are also being directed towards other value added products. Feruloyl esterases (EC. 3.1.1.73) can be used as a tool for bio-refining of lignocellulosic material for the recovery and purification of ferulic acid and related hydroxycinnamic acids ubiquitously found in the plant cell wall. More and more genes coding for feruloyl esterases have been mined out from various sources to allow efficient enzymatic release of ferulic acid and allied hydroxycinnamic acids (HCAs) from plant-based biomass. A sum up on enzymatic extraction of HCAs and its recovery from less explored agro residual by-products is still a missing link and this review brushes up the achieved landmarks so far in this direction and also covers a detailed patent search on this biomass refining enzyme.

Rapid mineralisation of the Organic Fraction of Municipal Solid Waste.

The effect of pH, C/N ratio, addition of a microbial consortium (MC) and temperature upon mineralisation of Organic Fraction of Municipal Solid Waste (OFMSW) was studied; mineralisation was measured through the CO2 production rate and total CO2 formation. Through this process up to 432.9mg of CO2g(-1) initial dry matter (IDM) after 2days of treatment was obtained. It was found that under a slightly acidic pH (5-6) and C/N of 30, the mineralisation process was accelerated. Moreover, temperature (27-50°C) had no effect on the total CO2 produced. The highest CO2 production rate (5.28d(-1)) was observed at 27°C, C/N ratio of 30 and 8% of microbial consortium; it is at least 3.52 times higher than that reported (1.5d(-1)). The highest release of reducing sugars was determined at 50°C, possibly due to an increase in hydrolytic enzymes. Results suggest the potential use of rapid mineralisation of OFMSW for further friendly environmental processes.

Mass transfer and hydrocarbon biodegradation of aged soil in slurry phase.

Addition of toluene into slurry phase laboratory microcosm is proposed in order to increase desorption rate of hydrocarbons and as an alternative to improve bioavailability of hydrocarbon in aged soils. Our studies showed that toluene has a positive effect on desorption of total petroleum hydrocarbons (TPH). Addition of 14,000 mg toluene/kg of soil, in highly polluted soil, increased the consumption rate of hydrocarbons three times in comparison to control without solvent. In 30 days the initial TPH concentration in soil, 292,000 mg/kg, diminished 45%. Although toluene was able to dissolve complex organic compounds such as asphaltene fraction, it probably yielded a highly toxic toluene-hydrocarbons phase. The inhibitory effect of toluene-TPH was also studied. A substrate inhibition model was used: the k(m) and k(i) constants were 57 and 490 mg TPH/L liquid phase, respectively. Experimental data were well described when the proposed model included sequential desorption and biodegradation phenomena. Damköhler number evaluation showed that rate of mass transfer was the limiting step in overall biodegradation in nonsolvent control. When high concentration of toluene was added, then bioreaction was the limiting step, but inhibitory effect should be considered. However, toluene addition at low concentrations facilitates the biodegradation of aromatic compounds.

Production of pectinases by Aspergillus niger in solid state fermentation at high initial glucose concentrations.

Exopectinase (exo-p) and endopectinase (endo-p) production by Aspergillus niger CH4 in solid state culture was studied at initial glucose concentrations of 100, 250, 350 and 450 g/l. The highest activity of exo-p (35 U/g) was produced at 72 and 120 h in the medium containing 100 and 250 g glucose/l, respectively. The maximum endo-p activity (9 U/g) was produced at 72 h in the medium with 250 g glucose/l. The reduction in pectinase production at 350 and 450 g/l initial glucose concentration was due neither to repression of the synthesis of the enzyme nor to the glucose consumption rate of the strain but due to a drastic drop in pH of the medium.

Symmetric branching model for the kinetics of mycelial growth.

A mathematical model, linking microscopic to macroscopic parameters of the kinetics of mycelial growth is presented. The model consists of two parts: (a) a microscopic description, based on the assumption that growth of a mycelium can be represented approximately by the growth of a symmetric binary tree, where the branching level (microscopic state variable) is logarithmically related to the number of tips and segments; and (b) a macroscopic description which makes use of the microscopic description in order to define the parameters related to the evolution of biomass (macroscopic state variable) as a function of time. The latter uses a distribution of arrested tips in a population of mycelia, in order to estimate the fraction of non-growing biomass in terms of a power law function with coefficient, n, of the biomass concentration. The microscopic description explains the fact that the germ tube specific growth rate of Aspergillus nidulans measured in a growth chamber, is about the double the specific growth rate of this organism, when measured in shake flasks. It predicts that the length of the hyphal growth unit of the mycelium of Geotrichum candidum would be approximately the double the germ tube length measured at the time just before the first branching event. It also allows the derivation of useful expressions for predicting macroscopic parameters, such as the maximal specific growth rate, the initial amount of biomass, and the amount of biomass before the branching process starts. Those estimates are done in terms of microscopic quantities, i.e., the amount of germinated spores, the diameters of the spores and hyphae, the average rate of tip extension, and the average internodal segment length. Estimation of coefficient n by fitting the macroscopic description to a growth curve of A. niger gives an indication on the degree of skewness of the distribution of arrested mycelia. Estimated macroscopic parameters are in relative good agreement with measured average segment length.

Importance of medium pH in solid state fermentation for growth of Schwanniomyces castellii.

Utilization of soluble starch by Schwanniomyces castellii in a solid state fermentation system was highest in unbuffered medium when initial and final pH of the medium were 6.5-7.0 and 4.0-4.6, respectively. An economic strategy involving the use of urea as a sole nitrogen source in medium with initial pH of 6.5 allowed maximum substrate utilization in the absence of buffer and without any contamination in column fermenter.

Heat transfer simulation in solid substrate fermentation.

A mathematical model was developed and tested to simulate the generation and transfer of heat in solid substrate fermentation (SSF). The experimental studies were realized in a 1-L static bioreactor packed with cassava wet meal and inoculated with Aspergillus niger. A simplified pseudohomogeneous monodimensional dynamic model was used for the energy balance. Kinetic equations taking into account biomass formation (logistic), sugar consumption (with maintenance), and carbon dioxide formation were used. Model verification was achieved by comparison of calculated and experimental temperatures. Heat transfer was evaluated by the estimation of Biot and Peclet heat dimensionless numbers 5-10 and 2550-2750, respectively. It was shown that conduction through the fermentation fixed bed was the main heat transfer resistance. This model intends to reach a better understanding of transport phenomena in SSF, a fact which could be used to evaluate various alternatives for temperature control of SSF, i.e., changing air flow rates and increasing water content. Dimensionless numbers could be used as scale-up criteria of large fermentors, since in those ratios are described the operating conditions, geometry, and size of the bioreactor. It could lead to improved solid reactor systems. The model can be used as a basis for automatic control of SSF for the production of valuable metabolites in static fermentors.