Abstract This manuscript reports the valorization of two-phase olive mill waste (TPOMW) as raw material and carbon source for solid state fermentation using Yarrowia lipolytica as biocatalyst. Due to its chemical characteristics, a combination of different raw materials (TPOMW and wheat bran, WB) was evaluated and two distinct nitrogen sources were applied as supplementation for lipase production. A TPOMW/WB ratio of 1:1 and supplementation with ammonium sulfate was chosen as the best condition. The productivity in 24 h reached 7.8 U/g*h and, after four days of process, only decreased about 35%. Process pH ranged from 5.5 - 5.9, remaining in an acid range. Thus, the successful use of TPOMW, a watery solid by-product with high content of lipids, as raw material for Yarrowia lipolytica growth and lipase production provided an environmental friendly alternative to valorize such waste.

Saponins, molecules classified as triterpenic or steroidal glycosides, are metabolites distributed in all the plant kingdom that can be used for the production of foods, cosmetics, and pharmaceuticals, as well as in soil bioremediation. Saponins are normally extracted from natural resources with water, ethanol and/or methanol, and then concentrated by liquid-liquid partitioning with n-butanol. An alternative concentration method is with a foam column, by which the saponins can be concentrated via preferential adsorption at a gas-liquid interface. Therefore, the objective of this work was the use of a foam column for the concentration of saponins from juá and sisal, evaluating parameters such as: initial working volume in the column, saponin concentration in the extracts from juá and sisal, air flow rate, pH, Raschig rings loading and operation time. When a gradient air flow rate and 25 g of Raschig rings were used, 82.6% of the jua saponins loaded onto the system were recovered in a 3.46-fold concentrated solution after 9 h of operation. Regarding sisal saponins, a concentration factor of 1.98 was observed with 90.5% of saponin recovery during 4.5 h of operation.

In this work, the production of a biosurfactant synthesized by Yarrowia lipolytica using different renewable resources as carbon source was investigated. Crude glycerol, a biodiesel co-product, and clarified cashew apple juice (CCAJ), an agroindustrial residue, were applied as feedstocks for the microbial surfactant synthesis. The microorganism was able to grow and produce biosurfactant on CCAJ and crude glycerol, achieving maximum emulsification indexes of 68.0% and 70.2% and maximum variations in surface tension of 18.0 mN.m-1and 22.0 mN.m-1, respectively. Different organic solvents (acetone, ethyl acetate and chloroform - methanol) were tested for biosurfactant extraction. Maximum biosurfactant recovery was obtained with chloroform - methanol (1:1), reaching 6.9 g.L-1for experiments using CCAJ and 7.9 g.L-1for media containing crude glycerol as carbon source.The results herein obtained indicate that CCAJ and the co-product of biodiesel production are appropriate raw materials for biosurfactant production by Y. lipolytica.

This work deals with the development of a digital image analysis procedure to characterize microbial flocs obtained in three different WWTP: a bench-scale Sequencing Batch Reactor (SBR) dealing with phenol and nitrogen biological removal, a municipal treatment unit (Ilha do Governador, Rio de Janeiro, Brazil) and an industrial wastewater treatment plant (Ciba - Estrada do Colégio, Rio de Janeiro, Brazil). The developed procedure permits to obtain its morphological parameters like equivalent diameter, compactness, roundness and porosity properties as well as the fractal dimension. This procedure was validated and lead to identify the major relationships between the analysed morphological parameters. A minimum of 300 flocs should be included in the image analysis and a significant influence of the sample dilution step on the mean size of the flocs was verified. The porosity parameter positively correlated with the fractal dimension of microbial aggregates indicating the that highly porous flocs are very irregular.

The methylotrophic yeast Pichia pastoris was used to produce the recombinant Pisum sativum defensin (rPsd1), a small peptide from pea seeds that has a high level of antifungal activity. The plasmid rPsd1/pPIC9 was integrated into the yeast genome and methanol was used to induce expression and secretion of the recombinant Psd1, at 30º C in a fed-batch mode. The effects of different pH conditions and process scale-up were evaluated using a Monod-type model where dissolved oxygen was considered the limiting substrate. Parameter estimation showed that the process could be improved by expressing rPsd1 in a 1000 mL bioreactor at pH 4. Structural and functional analyses revealed that the recombinant Psd1 is very similar to the native one.