A new year has come! And at the same time there is plenty of work that has to be done, now that all the materials neccesary are available. Our goals for this semester are:
- Successfully grow Phanerochaete chrysosporium in both rich and basal medium to observe non-granulated vs. pelletized cultures.
- Perform a protein expression screen to observe differential expression related to pelletization.
- Identify proteins that are being over-expressed or inhibited by the basal medium.
- Perform enzyme kinetics assays (if neccesary) to characterize unknown proteins.
- Explore which variables must be changed in order to reduce the size of the pellets to the optimum diameter in which the mass transfer and enzyme kinetics become efficient for possible industrial applications of filamentous fungi.
Last semester we were able to design the experimental setup, perform literature search, and ordering materials and equipment. The orders where received during the last weeks of the semester, thus, we are now well poised to perform the experimental setup. The initial steps of fungal growth and media preparation have already started and some weeks are needed to have a culture ready for media inoculation. After this steps, the observational and molecular assays will be perform to comply with the semester goals.
This semester we will be testing how the white rot fungi grows in the liquid media described by Jimenez-Tobón et al. (Enzyme and Microbial Technology, 1997). This involves digital imagery and measurement of pellet size. Also, the protein assays will be performed using SDS-PAGE to observe, by protein staining, the protein expression patterns of both rich and basal cultures. Identification of proteins of interest will be performed by LC-MS-MS in collaboration with the University of Wisconsin at Madison. Future tests will be performed in accordance with the initial results.
