In the last years, tannase has been the subject of a lot of studies due to its commercial importance and complexity as catalytic molecule. Tannases are capable of hydrolyzing complex tannins, which represent the main chemical group of natural anti-microbials occurring in the plants. The general outline of this work includes information of the substrates, the enzyme, and the applications. This review considers in its introduction the concepts and history of tannase and explores scientific and technological aspects. The "advances" trace the route from the general, molecular, catalytic, and functional information obtained under close to optimal conditions for microbial production through purification, description of the enzyme properties, and the commercial applications to the "perspectives" including expression studies, regulation, and potential uses; aspects related to the progress in our understanding of tannin biodegradation are also included.

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Structural and functional catalytic characteristics of cross-linked enzyme aggregates (CLEA) are reviewed. Firstly, advantages of enzyme immobilization and existing types of immobilization are described. Then, a wide description of the factors that modify CLEA activity, selectivity and stability is presented. Nowadays CLEA offers an economic, simple and easy tool to reuse biocatalysts, improving their catalytic properties and stability. This immobilization methodology has been widely and satisfactorily tested with a great variety of enzymes and has demonstrated its potential as a future tool to optimize biocatalytic processes.

Differential scanning calorimetry (DSC), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) were used to determine morphological, structural and surface changes (biodegradation) on thermo-oxidized (80°C, 15 days) low-density polyethylene (TO-LDPE) incubated with Aspergillus niger and Penicillium pinophilum fungi, with and without ethanol as cosubstrate for 31 months. TO-LDPE mineralization by fungi was also evaluated. Significantly morphological and structural final changes on biologically treated TO-LDPE samples were observed. Decreases to three units on crystallinity and crystalline lamellar thickness (0.4 -1.8 Å), and increases in small-crystals content (up to 3.2%) and mean crystallite size (8.4 -14 Å) were registered. An oxidation decrease (almost twice) on samples without ethanol with respect to the control was observed, while in those with ethanol it was increased (up to 2.5 times). Double bond index increased more than twice from 21 to 31 months. The higher TO-LDPE changes and fungi-LDPE interaction was observed in samples with ethanol, suggesting that ethanol favors the TO-LDPE biodegradation, at least in case of P. pinophilum, probably by means of a cometabolic process. Mineralization of 0.50 % and 0.57 % for A. niger, and of 0.64 % and 0.37 % for P. pinophilum were obtained, for samples with and without ethanol, respectively. A model to explain morphological and structural changes on biologically treated TO-LDPE is also proposed.

A study was made to compare the production of pectinase by Aspergillus niger CH4 in solid-state (SSF) and submerged (SmF) fermentations. Production of endo-(endo-p) and exo-pectinase (exo-p) by SSF was not reduced when glucose, sucrose or galacturonic acid (up to 10%0) were added to a culture medium containing pectin. Moreover, both activities increased when concentrations of the carbon sources were also increased. In SmF, these activities were strongly decreased when glucose or sucrose (3%) was added to culture medium containing pectin. The addition of galacturonic acid affected endo-p activity production to a lesser extend than exo-p. Final endo-p and exo-p activities in SSF were three and 11 times higher, respectively, than those obtained in SmF. The overall productivities of SSF were 18.8 and 4.9 times higher for endo-p and exo-p, respectively, than those in SmF. These results indicate that regulatory phenomena, such as induction-repression or activation-inhibition, related to pectinase synthesis by A. niger CH4 are different in the two types of fermentation.

The performance of biofilters inoculated with the fungus Scedosporium apiospermum was evaluated. This fungus was isolated from a biofilter which operated with toluene for more than 6 months. The experiments were performed in a 2.9 L reactor packed with vermiculite or with vermiculite-granular activated carbon as packing material. The initial moisture content of the support and the inlet concentration of toluene were 70% and 6 g/m 3 , respectively. As the pressure drop increased from 5-40 mm H 2 O a strong initial growth was observed. Stable operation was maintained for 20 days with a moisture content of 55% and a biomass of 33 mg biomass/g dry support. These conditions were achieved with intermittent addition of culture medium, which permitted a stable elimination capacity (EC) of 100 g/m 3 reactor h without clogging. Pressure drop across the bed and CO 2 production were related to toluene elimination. Measurement of toluene, at different levels of the biofilter, showed that the system attained higher local EC (200 g/m 3 r h) at the reactor outlet. These conditions were related to local humidity conditions. When the mineral medium was added periodically before the EC decreases, EC of approximately 258 g/m 3 r h were maintained with removal efficiencies of 98%. Under these conditions the average moisture content was 60% and 41 mg biomass/g dry support was produced. No sporulation was observed. Evaluation of bacterial content and activities showed that the toluene elimination was only due to S. apiospermum catabolism.

Tannase production by Aspergillus niger Aa-20 was studied in submerged (SmF) and solid-state (SSF) fermentation systems with different tannic acid and glucose concentrations. Tannase activity and productivity were at least 2.5 times higher in SSF than in SmF. Addition of high tannic acid concentrations increased total tannase activity in SSF, while in SmF it was decreased. In SmF, total tannase activity increased from 0.57 to 1.03 IU/mL, when the initial glucose concentration increased from 6.25 to 25 g/L, but a strong catabolite repression of tannase synthesis was observed in SmF when an initial glucose concentration of 50 g/L was used. In SSF, maximal values of total tannase activity decreased from 7.79 to 2.51 IU when the initial glucose concentration was increased from 6.25 to 200 g/L. Kinetic results on tannase production indicate that low tannase activity titers in SmF could be associated to an enzyme degradation process which is not present in SSF. Tannase titers produced by A. niger Aa-20 are fermentation system-dependent, favoring SSF over SmF.

In the last years, tannin biodegradation has been the subject of a lot of studies due to its commercial importance and scientific relevance. Tannins are molecules of low biodegradation and represent the main chemical group of natural anti-microbials occurring in the plants. Among the different kinds of tannins, ellagitannins represent the group less studied manly due to their diversity and chemical complexity. The general outline of this work includes information on tannins, their classification and properties, biodegradation, ellagic acid production, and potential applications. In addition, it describes molecular, catalytic, and functional information. Special attention has been focused on the biodegradation of ellagitannins describing the possible role of microbial enzymes in the production of ellagic acid.