Determining the Presence and Location of Recombinant Pichia pastoris Cells and Green Fluorescent Protein in the Digestive Tract of Mice
Faculty Mentor Name
Der Thor
Research or Creativity Area
Natural Sciences
Abstract
Pichia pastoris, well known for its high efficiency in producing recombinant proteins, is a species of yeast that is widely used in biotechnology and pharmaceutical research. The goal of this study is to determine whether P. pastoris can colonize and secrete recombinant proteins in the murine gut. Mice were fed this yeast in the form of freeze-dried dough made of standard rodent chow and concentrated P. pastoris. Sections of gastrointestinal tissue samples were collected at 0, 24, 48, and 96 hours post-feeding. Luminal content from these sections was plated on selective media to quantify viable P. pastoris colonies. Enzyme-linked immunosorbent assay (ELISA) was then performed to determine green fluorescent protein (GFP) concentrations.
Results demonstrated that both GFP and P. pastoris were most abundant in the 0-hour collection time. GFP was primarily detected in the cecum and colon, while P. pastoris colonies were most concentrated in the cecum. At the 24 hour collection time, these values were maintained at relatively high levels. At the 48 and 96 hour collection times, both P. pastoris colony counts and GFP concentrations were substantially lower compared to the 0 hour collection time. Overall, the decline in GFP levels correlated with the reduction in viable P. pastoris colonies, showing a direct relationship between yeast presence and protein production.
This research demonstrated that the recombinant P. pastoris is able to survive in the gastrointestinal tract and secrete detectable proteins at significant levels early after consumption. These results suggest that P. pastoris has potential use in future therapeutic applications, such as treating lactose intolerance.
Determining the Presence and Location of Recombinant Pichia pastoris Cells and Green Fluorescent Protein in the Digestive Tract of Mice
Pichia pastoris, well known for its high efficiency in producing recombinant proteins, is a species of yeast that is widely used in biotechnology and pharmaceutical research. The goal of this study is to determine whether P. pastoris can colonize and secrete recombinant proteins in the murine gut. Mice were fed this yeast in the form of freeze-dried dough made of standard rodent chow and concentrated P. pastoris. Sections of gastrointestinal tissue samples were collected at 0, 24, 48, and 96 hours post-feeding. Luminal content from these sections was plated on selective media to quantify viable P. pastoris colonies. Enzyme-linked immunosorbent assay (ELISA) was then performed to determine green fluorescent protein (GFP) concentrations.
Results demonstrated that both GFP and P. pastoris were most abundant in the 0-hour collection time. GFP was primarily detected in the cecum and colon, while P. pastoris colonies were most concentrated in the cecum. At the 24 hour collection time, these values were maintained at relatively high levels. At the 48 and 96 hour collection times, both P. pastoris colony counts and GFP concentrations were substantially lower compared to the 0 hour collection time. Overall, the decline in GFP levels correlated with the reduction in viable P. pastoris colonies, showing a direct relationship between yeast presence and protein production.
This research demonstrated that the recombinant P. pastoris is able to survive in the gastrointestinal tract and secrete detectable proteins at significant levels early after consumption. These results suggest that P. pastoris has potential use in future therapeutic applications, such as treating lactose intolerance.
