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Recombinant Saccharomyces cerevisiae expressing P450 in artificial digestive systems : A model for biodetoxication in the human digestive environment

Author: Blanquet, S. · Meunier, J.P. · Minekus, M. · Marol-Bonnin, S. · Alric, M.
Type:article
Date:2003
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Applied and Environmental Microbiology, 5, 69, 2884-2892
Identifier: 237078
doi: doi:10.1128/AEM.69.5.2884-2892.2003
Keywords: Health Nutrition · Physiological Sciences · Digestive system · Disease control · Microbiology · Microorganisms · Biodrugs · Yeast · Cinnamate 4 monooxygenase · Cinnamic acid · cytochrome P450 · Oxygenase · Para coumaric acid · Unclassified drug · Biotechnology · Drug development · Microbiology · Article · Biotransformation · Computer simulation · Detoxification · Digestion · Digestive system · Duodenum · Enzyme activity · Gastrointestinal tract · Ileum · Incubation time · Intestine secretion · Jejunum · Quantitative analysis · Saccharomyces cerevisiae · Small intestine · Stomach secretion · Survival rate · Yeast · Genetic Engineering · Helianthus · Humans · Metabolic Detoxication, Drug · Mixed Function Oxygenases · Models, Biological · Saccharomyces cerevisiae · Trans-Cinnamate 4-Monooxygenase · Saccharomyces cerevisiae

Abstract

The use of genetically engineered microorganisms such as bacteria or yeasts as live vehicles to carry out bioconversion directly in the digestive environment is an important challenge for the development of innovative biodrugs. A system that mimics the human gastrointestinal tract was combined with a computer simulation to evaluate the survival rate and cinnamate 4-hydroxylase activity of a recombinant model of Saccharomyces cerevisiae expressing the plant P450 73A1. The yeasts showed a high level of resistance to gastric and small intestinal secretions (survival rate after 4 h of digestion, 95.6% ± 10.1% [n = 4]) but were more sensitive to the colonic conditions (survival rate after 4 h of incubation, 35.9% ± 2.7% [n = 3]). For the first time, the ability of recombinant S. cerevisiae to carry out a bioconversion reaction has been demonstrated throughout the gastrointestinal tract. In the gastric-small intestinal system, 41.0% ± 5.8% (n = 3) of the ingested transcinnamic acid was converted into p-coumaric acid after 4 h of digestion, as well as 8.9% ± 1.6% (n = 3) in the stomach, 13.8% ± 3.3% (n = 3) in the duodenum, 11.8% ± 3.4% (n = 3) in the jejunum, and 6.5% ± 1.0% (n = 3) in the ileum. In the large intestinal system, cinnamate 4-hydroxylase activity was detected but was too weak to be quantified. These results suggest that S. cerevisiae may afford a useful host for the development of biodrugs and may provide an innovative system for the prevention or treatment of diseases that escape classical drug action. In particular, yeasts may provide a suitable vector for biodetoxication in the digestive environment. Chemicals/CAS: cinnamate 4 monooxygenase, 9077-75-2; cinnamic acid, 4151-45-5, 538-42-1, 621-82-9; cytochrome P450, 9035-51-2; oxygenase, 9037-29-0, 9046-59-7; para coumaric acid, 7400-08-0; Cytochrome P-450 Enzyme System, 9035-51-2; Mixed Function Oxygenases, EC 1.-; Trans-Cinnamate 4-Monooxygenase, EC 1.14.13.11