Studies of the Proteolytic Activity of the Subcellular Particulates of the Cells of the Leaves of Spinacia oleracea, L.

111 Pages·4.291 MB·English

by  Alscher, Ruth Paula

FORDHAM UNIVERSITY
GRADUATE SCHOOL
May 15 * 19.51
This dissertation prepared under my direction by
Ruth Paula Alscher
entitled STUDIES OF THE PROTEOLYTIC ACTIVITY
OF THE SUBCELLUIAR PARTICULATES OF THE
.......................................CELLS OF THE IEAVES OF SPBTACIA OLBRACSA,L.
has been accepted in partial fulfilment of the requirements for the
Degree of Doctor of Philosophy
Dr. Peter Rieser
(Faculty Adviser)

r
STUDIES ON THE PROTEOLYTIC ACTIVITY
OF THE SOBCELLULAR PARTICULATES OF THE CELLS
OF THE LEAVES OF SPINACIA OLERACEA, L.
BY
RUTH PAULA ALSCHER
A*B*, College of New Rochelle, *45
M.S*, Fordham University, *46
DISSERTATION
SUBMITTED IN PARTIAL fULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY
IN THE DEPARTMENT OF BIOLOGY AT
FORDHAM UNIVERSITY
NEW YORK
1951
L
ProQuest Number: 10992971
All rights reserved
INFORMATION TO ALL USERS
The quality of this reproduction is dependent upon the quality of the copy submitted.
In the unlikely event that the author did not send a complete manuscript
and there are missing pages, these will be noted. Also, if material had to be removed,
a note will indicate the deletion.
uest
ProQuest 10992971
Published by ProQuest LLC(2018). Copyright of the Dissertation is held by the Author.
All rights reserved.
This work is protected against unauthorized copying under Title 17, United States Code
Microform Edition © ProQuest LLC.
ProQuest LLC.
789 East Eisenhower Parkway
P.O. Box 1346
Ann Arbor, Ml 48106- 1346

iii
r
TABLE OF CONTENTS
Page
LIST OF TABLES................................................................. v
LIST OF FIGURES.............................. vi
ACKNOWLEDGMENTS ....................................... v iii
I. INTBODUCTION..................................................... 1
II. MATERIAL AND METHODS . .......................................... 8
III. OBSERVATIONS AND RESULTS......................................... 25
Spinach leaf structure • • • • • • ........................ 25
The effect of pressure on the cells and
the subcellular particulates. • • • • . • • • 31
Spinach brei treated with ether.......................... 31
The cells treated with sodium chloride
solution ........................................................... 33
The spinach b r e i • • • • 35
Spinach filtrates .............................................. . . . 35
The proteolytic activity of frozen
spinach • * • • • * • • « . • • • • » • • • • 36
The effect of storage of spinach filtrates
on proteolytic a ctiv ity ................................... 39
The proteolytic activity of identical
spinach preparations................................................. 39
The proteolytic activity of summer spinach
and winter spinach. ............................................ 44
Spinach preparations centrifuges for
5 minutes . . . 44
Spinach preparations centrifuged for
10 m inutes........................ 51
Proteolytic activity of the yellow-green
and light green granules. • » • • • • • • • • 54
Spinach preparations centrifuged for
20 m inutes................................ 55
Spinach preparations after centrifugation
for 40 minutes.................................. 61
The effect of various incubation periods
on the rate of digestion of casein • • • • • 64
The effect of pH on the pro­
teolytic activity of spinach filtrates. . . . 66
iv
r
Page
IV. DISCUSSION........................................................................... 71
V. SUMMARY.................................................................................. 86
BIBLIOGRAPHY.............................. 90
L
V
r n
LIST OF TABLES
Table Page
I, Measurements of the subcellular
particulates of the cells of the
leaves of Spinacia oleracea . . . . . . . . . SO
II* The proteolytic activity of
identical preparations of
spinach filtrates ......................................... 43
III. Proteolytic activity of summer
and winter sp in ach ......................................................... 45
IV. Percent of digestion of
spinach filtrates incubated
for various periods of tim e..................................... 65
L
vi
r n
LIST OF FIGURES
Figure Page
1* A cross-section view of two
palisade cells ................................ . . . . . . 27
2* A cross-section view of a large
spongy c e ll....................• . 27
5* A cross-section view of smaller
spongy cells ....................................... . . . . 27
4* A surface view of two guard cells
on the upper epidermis .................................... 27
5. Yell ow-green and light green
granules, fragments of chloroplasts
and whole chloroplasts within a
spongy c e ll................................................................. 29
6* Chloroplasts, clumps of chloroplasts
and yellow-green and light green
granules isolated from the cells . ♦ • . . 29
7* A cross-section view of spongy
cells prior to the application of
pressure .................................... . . . . . . . . 29
8* A cross-section view of the same
spongy cells after the application
of pressure .............................. 29
9* Proteolytic activity of filtrates
of fresh and frozen spinach • • • • • . . 38
10. Proteolytic activity of fresh
filtrate and stored filtrate of
spinach • • • « • • • • • • • ....................... 41
11. Proteolytic activity of summer and
winter spinach ................................................. 47
12. Proteolytic activity of spinach
preparations after 5 minutes of
centrifugation.......................................................... 50
vli
Page
15. Proteolytic activity of spinach
preparations after 10 minutes of
centrifugation • • • • • • • • • . . .................. 55
14. Proteolytic activity of yellow-green
and light green granules • • • • • ....................... 57
15. Proteolytic activity of spinach pre­
parations after 20 minutes of
centrifugation .......................... 60
16. Proteolytic activity of clear super­
natant fluid after 40 minutes of
centrifugation .............................. 65
17* Proteolytic activity of spinach
filtrates at various hydrogen ion
concentrations I . . . . . ........................................ 68
18. Proteolytic activity of spinach
filtrates at various hydrogen ion
concentrations II . . . .................. . . . . . . 70
L