Spoilage yeasts in the wine industry.

Recombinant protein production is a multibillion-dollar market. The development of a new product begins with the choice of a production host. While one single perfect host for every protein does not exist, several expression systems ranging from bacterial hosts to mammalian cells have been established. Among them, yeast cell factories combine the advantages of being single cells, such as fast growth and easy genetic manipulation, as well as eukaryotic features including a secretory pathway leading to correct protein processing and post-translational modifications. In this respect, especially the engineering of yeast glycosylation to produce glycoproteins of human-like glycan structures is of great interest. Additionally, different attempts of cellular engineering as well as the design of different production processes that are leading to improved productivities are presented. With the advent of cheaper next-generation sequencing techniques, systems biotechnology approaches focusing on genome scale analyses will advance and accelerate yeast cell factories and thus recombinant protein production processes in the near future. In this review we summarize advantages and limitations of the main and most promising yeast hosts, including Saccharomyces cerevisiae, Pichia pastoris, and Hansenula polymorpha as those presently used in large scale production of heterologous proteins.

Recombinant Protein Production in Yeasts

Recombinant DNA (rDNA) technologies (genetic, protein, and metabolic engineering) allow the production of a wide range of peptides, proteins, and biochemicals from naturally nonproducing cells. These technologies, now approx 25 yr old, have become one of the most important technologies developed in the twentieth century. Pharmaceutical products and industrial enzymes were the first biotech products on the world market made by means of rDNA. Despite important advances in rDNA applications in mammalian cells, yeasts still represent attractive hosts for the production of heterologous proteins. In this review we summarize advantages and limitations of the main and most promising yeast hosts.

Recombinant protein production in yeasts.

One stop mycology

UNLABELLED This experimental work was aimed to evaluate the effects of repeated high-pressure homogenization (HPH) treatments at 100 MPa on the inactivation and regrowth of Zygosaccharomyces bailii inoculated in apricot and carrot juices. Thus, the spoilage yeast was inoculated in both the juices at level of about 5 log CFU/g and the 2 systems were treated with a lab-scale Panda homogenizer for 8 passes at 100 MPa. Microbiological and chemico-physical analyses were performed immediately after the treatment and during the juice storage at room temperature. Microbial data highlighted that yeast inactivation increased with the number of passes applied. Eight passes at 100 MPa allowed yeast inactivation higher than 2.5 log CFU/mL regardless of the juice considered. On the contrary, the juice type affected the yeast fate (growth or death) over the storage at 25 degrees C. In fact, Z. bailii was able to attain the spoilage threshold (6 log CFU/mL) in apricot juice, although with growth kinetics dependent of the survivor levels after HPH treatment. In carrot juice this microorganism was unable to recover over the storage in the most severely treated samples. The HPH treatment had a significant effect on apricot juice pH and viscosity, while no significant effect was observed in carrot juice. The viscosity measurements showed that the application of one pass at 100 MPa resulted in the triplication of apricot viscosity index. No further significant viscosity increase (P > 0.05) was observed increasing the number of passes at 100 MPa. PRACTICAL APPLICATION The results obtained in the present study and the proposed technology could be exploited by the industries of the beverage sector to increase the shelf life of these kinds of products. Moreover, from a technological point of view, the increase of viscosity, following the high-pressure homogenization treatment, represents a tool to expand the product gamma without the use of gelling additives or thermal treatments, which are detrimental for the sensorial and nutritional properties of this kind of products.

Adel K. Makdesi

Papers

Evaluation of media for enumerating heat-stressed, benzoate-resistant Zygosaccharomyces bailii.