Premium
On‐going Studies on the Structural Stability of E. coli Alkaline Phosphatase by an Undergraduate Biochemistry Laboratory Course
Author(s) -
Hazzard James T.,
Schnoebelen Carly
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.613.5
Subject(s) - denaturation (fissile materials) , chemistry , alkaline phosphatase , tcep , biochemistry , enzyme , chromatography , nuclear chemistry , phosphine , catalysis
Our biochemistry laboratory course engages in a guided‐inquiry exercise in which we study the high degree of structural stability of E. coli alkaline phosphatase (AP) and the roles of two disulfide bonds toward thermal and chemical denaturation. Recently we have compared the denaturation of the enzyme using guanidinium hydrochloride and urea, in the absence and presence of the reducing agent TCEP, using CD and fluorescence spectroscopic techniques. We have also investigated the effect of temperature on the chemical denaturation processes. We also have continued to determine the nature of a chaperonin‐like protein(s) found in the periplasmic lysates that contributes to the dramatic thermal stability of AP, relative to the highly purified commercial enzyme using tandem mass spectrometry. Being a portion of an undergraduate biochemistry laboratory course, these investigations represent data obtained from individual laboratory groups who make oral presentations at the end of their research, providing a more real world approach to the educational experience. This work is supported, in part, by a grant from NSF (DUE 0837398).