Glucaric acid, my product of interest, overlaps with phosphate present in the media in the HPLC column HPX-87H (biorad). So, I was wondering if there are other known phosphorus sources that I could use.
Hello Wesley!
AFAIK - phosphates would be the only form of phosphorous consumable by microorganisms from growth media.
What could be an options to solve the issue:
1) try another column / method (unless you are following some USP / EP procedures or other standard methods)
2) precipitation of phosphate from sample and keeping the glucaric acid in solution before analysis
3) extraction of glucaric acid into other solvent and analyze the extract
Hope it will help.
Hi Wesley,
Would glycerophosphoinositol be a valid alternative? S. cerevisiae is able to use this compound as phosphate source via import and further intracelular degradation (1). I don't know if it overlaps with glucaric acid in the 87H column, but it's an alternative phosphate source for yeast.
1: Article Production and reutilization of an extracellular phosphatidy...
The extracellular phosphatases of yeast have a very broad substrate specificity. From reading the literature glycerophosphocholine should work. Inositols will not work in yeast because they are missing the phytases. Maybe it is also worth to check beta-glycerophosphate, which is a cheap alternative to glycerophosphocholine.
- Badges
- Science method