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From 2008.igem.org

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External links to articles used to formulate optimization of gene regulation and overall process efficency:

Missouri S&T Biological Sciences; Missouri S&T Chemical and Biological Engineering; Missouri S&T Homepage; SDSC Biology Workbench; NCBI; ATCC

The following papers describe characterization of alcohol oxidase activity in response to ethanol and formed the basis for selecting the AOX1 promoter region from Pichia pastoris.

A complex pathway for the metabolism of methanol exists within some species of the Pichia genus. Alcohol oxidase (AO) appears to be the first and major enzyme produced in this metabolic pathway (1). Transcribed from its gene (AOX1), AO converts methanol to formaldehyde within the yeast’s peroxisome (1). A metabolic pathway for the utilization of ethanol is also present within the yeast. However, if both ethanol and methanol is present, the yeast will utilize the ethanol before consuming the methanol (2). Consequently, the AOX gene will not be expressed to produce the AO enzyme until the ethanol has been consumed.

1. Cregg, James M., K. R. Madden, K. J. Barringer, G. P Thill, and C. A. Stillman. 1989. Functional Characterization of the Two Alcohol Oxidase Genes from the Yeast Pichia pastoris. Molecular and Cellular Biology. 9:1316-1323.

2. Inan Mehmet and Michael M. Meagher. The Effect of Ethanol and Acetate on Protein Expression in Pichia pastoris. 2001. Journal of Bioscience and Bioengineering. 9: 337-341.

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