L-ascorbic acid producing yeasts learn from plants how to recycle it.

Microorganisms employed in industrial fermentation processes are often subjected to a variety of stresses that negatively affect growth, production and productivity. Therefore, stress robustness is an important property for their application. Reactive Oxygen Species (ROS) accumulation is a common denominator to a lot of these stress factors. Ascorbic acid (L-AA) acts as ROS scavenger, thus potentially protecting cells from harmful oxidative products. We have previously reported the development of Saccharomyces cerevisiae strains able to produce L-AA. This was obtained by expressing the known plant pathway genes and by complementing the missing step with an animal activity. Here, we show that L-AA accumulation inside yeast cells can be improved by expressing the complete biosynthetic plant pathway and, even further, by recycling its oxidation products. These new strains can be seen in a double perspective of exploitation: as novel organisms for vitamin C production and as novel cell factories for industrial processes.

E40, a novel microbial protease efficiently detoxifying gluten proteins, for the dietary management of gluten intolerance.

Gluten proteins are the causative agent of Celiac Disease (CD), a life-long food intolerance characterized by an autoimmune enteropathy. Inadvertent gluten exposure is frequent even in celiac patients complying with a gluten-free diet, and the supplementation of exogenous gluten-digestive enzymes (glutenases) is indeed a promising approach to reduce the risk of dietary gluten boost. Here we describe Endopeptidase 40, a novel glutenase discovered as secreted protein from the soil actinomycete Actinoallomurus A8, and its recombinant active form produced by Streptomyces lividans TK24. E40 is resistant to pepsin and trypsin, and active in the acidic pH range 3 to 6. E40 efficiently degrades the most immunogenic 33-mer as well as the whole gliadin proteins, as demonstrated by SDS-PAGE, HPLC, LC-MS/MS, and ELISA. T lymphocytes from duodenal biopsies of celiac patients showed a strongly reduced or absent release of IFN-γ when exposed to gluten digested with E40. Data in gastrointestinal simulated conditions suggest that no toxic peptides are freed during gluten digestion by E40 into the stomach to enter the small intestine, thus counteracting the intestinal inflammatory cascade to occur in CD patients. E40 is proposed as a novel candidate in Oral Enzymatic Therapy for the dietary management of gluten toxicity.