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Wheat invertases : characterization of cell wall-bound and soluble forms
H B Krishnan1, J T Blanchette, T W Okita
1Institute of Biological Chemistry and Graduate Program in Plant Physiology, Washington State University, Pullman, Washington 99164-6340.
Wheat coleoptiles contain two invertase forms: soluble and cell wall-bound. Light increases cell wall invertase activity, but not soluble invertase.
Area of Science:
- Biochemistry
- Plant Physiology
Background:
- Wheat coleoptiles possess two distinct invertase enzymes: soluble and cell wall-bound.
- These enzymes exhibit differences in pH optima, chromatography, kinetics, thermal stability, and light response.
Purpose of the Study:
- To characterize the soluble invertase from wheat coleoptiles.
- To investigate the effect of light on invertase activity in wheat.
Main Methods:
- Purification of soluble invertase using diethylaminoethyl-cellulose, concanavalin-A Sepharose, and Sephacryl S-300 chromatography.
- Enzyme characterization through polyacrylamide gel electrophoresis under denaturing conditions.
- Assessing invertase activity changes in response to light treatment.
Main Results:
- Soluble invertase was purified 175-fold with 26% recovery.
- The holoenzyme molecular weight was estimated at 158,000, with subunits at 53,000.
- Light illumination increased cell wall invertase activity but did not affect soluble invertase activity.
Conclusions:
- Wheat coleoptiles have biochemically distinct soluble and cell wall-bound invertases.
- Light specifically modulates cell wall-bound invertase activity in wheat coleoptiles.
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