Although esterases have been reported in peas, their properties
have not been studied. The purpose of this investigation was to
determine the substrate and inhibitor specificity of the esterases
present in a water extract of lyophilized peas, and to determine if
pea esterases could be classified according to the criteria established
for animal esterases.
Esterase activity was determined manometrically using the
Gilson differential respirometer. The effect of pH on esterase
activity was determined using phenyl acetate, phenyl propionate,
tripropionin, and tributyrin as substrates. The pH optima appeared
to lie between 6.9 and 7.2, and pH 7 was selected for use in this
study. The acetyl, propionyl, and n-butyryl esters of phenol,
2-naphthol-6-SO₃ Na and glycerol were hydrolyzed by the enzyme
extract. Long chain esters of 2-naphthol-6-SO₃Na, however, were not hydrolyzed. Cholinesterases and lipases did not appear to be
present in the extract since only a very small amount of activity
was observed when the choline esters and triolein were used as
substrates.
Using phenyl propionate and phenyl butyrate as substrates,
esterase activity, based on the original extract, decreased with
dilution. Later work revealed that the esterase(s) which hydrolyzed
phenyl propionate were inhibited by heavy metal ions and activated
by metal complexing agents. Hence, a possible explanation for the
decrease in activity was inhibition by metal ions in the distilled
water.
The effects of the inhibitors parathion, tetraethyl pyrophosphate,
and diisopropyl phosphorofluoridate at concentrations ranging from 10⁻¹ M to 10⁻¹⁰ M on esterase activity were determined.
The data suggested the presence of as many as six esterases in the
aqueous extract of peas, three for which the evidence was quite conclusive.
On the basis of their inhibition by organophosphorus compounds,
all but one of the esterases appeared to be of the B type.
Physostigmine sulfate (10⁻⁵ M) had no effect on esterase
activity with the nine substrates used indicating that the activity was
not due to cholinesterases.
At least one of the esterases which hydrolyzed each of the
substrates was sensitive to 10⁻³ M p-chloromercuribenzoic acid sodium salt suggesting the importance of sulfhydryl groups for
enzyme activity. / Graduation date: 1968
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/26801 |
| Date | 11 August 1967 |
| Creators | Norgaard, Marida Jane |
| Contributors | Montgomery, Morris W. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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