A perspective on the use of Pleurotus for the development of convenient fungi-made oral subunit vaccines.

This review provides an outlook of the medical applications of immunomodulatory compounds taken from Pleurotus and proposes this fungus as a convenient host for the development of innovative vaccines. Although some fungal species, such as Saccharomyces and Pichia, occupy a relevant position in the biopharmaceutical field, these systems are essentially limited to the production of conventional expensive vaccines. Formulations made with minimally processed biomass constitute the ideal approach for developing low cost vaccines, which are urgently needed by low-income populations. The use of edible fungi has not been explored for the production and delivery of low cost vaccines, despite these organisms' attractive features. These include the fact that edible biomass can be produced at low costs in a short period of time, its high biosynthetic capacity, its production of immunomodulatory compounds, and the availability of genetic transformation methods. Perspectives associated to this biotechnological application are identified and discussed.

Coupling aerobic biodegradation of methanol vapors with heterologous protein expression of endochitinase Ech42 from Trichoderma atroviride in Pichia pastoris.

Methanol is included among the most hazardous air pollutants, and an effort of vapors biofiltration by using microbial consortiums has been reported. The aim of this work was to couple the methanol vapors biodegradation with the production of recombinant endochitinase (ech42) from Trichoderma atroviride in Pichia pastoris transformed with the pPIC-ech42 plasmid. After carrying out batch experiments at 0.5% (w/v) of methanol concentration, the recombinant P. pastoris Mut(+) strain was selected because it showed methanol biodegradation rates similar to those of wild type GS115 strain (39 g/m(3)h), but 15% higher than the transformed Mut(S) strain. In addition, the recombinant Ech42 protein production was higher in Mut(+) than Mut(S). After various methanol vapor concentrations were evaluated, the maximum recombinant protein recovery was 317 mg/l and the volumetric methanol consumption rate was 88.7 g/m(3)h at 0.5% (w/v) of methanol concentration. This research underlines the promising application of linking methanol vapors biodegradation with the production of recombinant protein with high biotechnological interests.

Overexpression, purification and characterization of the Trichoderma atroviride endochitinase, Ech42, in Pichia pastoris.

The endochitinase gene ech42 from Trichoderma atroviride was cloned and expressed in Pichia pastoris using a constitutive expression system. Over 98% of the recombinant protein was secreted into the culture medium as glycoprotein. A high endochitinase concentration, 186 mg/L with a specific enzyme activity of 14,128 Umg(-1) was produced. The optimal enzyme kinetic parameters for the recombinant protein were identical to those reported for the enzyme isolated from T. atroviride. The recombinant endochitinase possesses suitable features for biotechnological applications, such as activity at acidic pH and thermostability.