Oxidative Metabolism of Glucose by Penicillium chryogenum Q-176

110 Pages·10.513 MB·English

by  Stout, Helen Anne

PURDUE UNIVERSITY
THIS IS TO CERTIFY THAT THE THESIS PREPARED UNDER MY SUPERVISION
by H e le n A nne S t o u t _______________________________________________
e n title d O x id a ti v e me ta b D li fm _ o f _g] n cm se b y P é n ic illiu m
ch ry so g en u m 0-176___________________________________________________________
COMPLIES WITH THE UNIVERSITY REGULATIONS ON GRADUATION THESES
AND IS APPROVED BY ME AS FULFILLING THIS PART OF THE REQUIREMENTS
FOR THE DEGREE OF
Doctor of Philo.sQphg:
lOfessqr in Charge of Thesis
Head of School or Department
19
TO THE LIBRARIAN:----
*Sr-
THIS THESIS IS NOT TO BE REGARDED AS CONFIDENTIAL.
©HAD. SCHOOZi POKM 9—3.«0—Uf

OXIDATIVE METABOLISM OF
GLUCOSE BY PENICILLIUM CHRYSOGENUM Q-176
A Thesis
Submitted to the Faculty
of
Purdue University
by
Helen Anne Stout
In Partial Fulfillment of the
Requirements for the Degree
of
Doctor of Philosophy
September, 1950
ProQuest Number: 27714130
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ACKNOWLEDGMENT
I appreciate deeply the encouragement, stimulation, and
guidance provided by Dr. Henry Koffler throughout this work. Further­
more, I am indebted to the Bristol Laboratories and the National
Institutes of Health for fellowships which enabled me to do this
researche
TABLE OF CONTENTS
Page
ABSTRACT --------------------------------------------------------
INTRODUCTION----------------------------------------------------- 1
REVIEW OF THE LITERATURE------------------------------------------ 3
Oxidative Assimilation of Carbon by Fungi ----------------- 3
Effect of Azide and 2,4-Dinitrophenol upon Metabolism----------- 3
METHODS---------------------------------------------------------- 10
Preparation of Spore Stocks— —— — —— —— ^— — — ———— —— — ——— '— 10
Preparation of Cells---------------------------------- 10
Method of Determining Assimilation— ------------------- 11
Manometric Procedures ------------------ — ----------—-— --- 12
Chemical Analyses— ------- 13
Fate of the Endogenous Respiration during the Utilization of
Glucose---------------------------------- 14
Effect of Various Factors upon Oxidative Assimilation------------ 17
A. Mincing of the Cells---------------------------------- 17
B. Concentration of Glucose in the CultureM edium -----— 19
C. Ratio of Glucose to Ammonium Nitrate in theC ulture Medium 20
D* Concentration of Glucose '■'■■■ ™ ..■ » ». «mi ■mi . , , i. ,u , 21
E# pH------ — ---- 24
F. Starvation of the Cells-------------------------------- 26
G. Age of the Cells--------------------------------------- 28
Sodium Azide- — ---------------------------------- 30
Fermentative Assimilation------------------------------------- 32
Nature of the Assimilated Product------------------------------ 33
A. Cytochemical Evidence----------------------------------- 33
B. Acid Hydrolysis of Dried Cells---------- --------------- 36
Equation for the Utilization of Glucose------------------------ 38
Page
Ability of a Number of Strains of the Pénicillium notatum-
chrysogenum group to Assimilate Glucose— — — — — —— — — —— 1+0
GENERAL DISCUSSION------------------------------------------------ 42
SUMMARY AND CONCLUSIONS— — — — — — — — — — — — — — — — — — — — — 4S
RBFjSEtLNCES— —— - ——i< .i— »i...—i 61
VITA------------------------------------------------------------- 65
LIST OF TABLES
Table Page
1 Per cent glucose oxidized by P» chrysogenum Q—176 when
various concentrations were utilized—— --— — — — — ---- 15
2 Per cent glucose oxidized by P^ chrysogenum Q—176 when
various concentrations were utilized------ — ---— --- 16
3 Effect of mincing time upon the oxidation of glucose by
P* chrysogenum Q—176— — — 18
4 Effect of concentration of glucose in the culture medium
upon the oxidative assimilation of glucose by P^
5 Effect of the glucose to ammonium nitrate ratio of the
culture medium upon the oxidative assimilation of
glucose by P * chrysogenum Q—176— — — — — — — 21
6 Effect of glucose concentration upon the oxidative
assimilation of glucose by P*_ chrysogenum Q-176-------- 22
7 Per cent glucose carbon assimilated by P^ chrysogenum Q-176
at various concentrations of glucose — 23
8 Change in pH after incubation of resting cells of P*
chrysogenum Q-176 with glucose for five hours-— -----— 24
9 Comparison between values obtained when carbon was
determined and those obtained when it was calculated— — 26
10 Effect of pH upon the oxidative assimilation of glucose by
11 Per cent glucose carbon assimilated by P^ chrysogenum Q-176
at various pH values— — — — — — — — — — — — — — — 27
12 Fermentative assimilation of glucose by chrysogenum Q-176 33
13 Increase in reducing sugar upon acid hydrolysis of dried
cells of P^ chrysogenum Q-176------------------------- 37
14 Extent of oxidative assimilation of glucose by cells of
P. chrysogenum Q-176 subjected to acid hydrolysis-— -- 37
15 Increase in reducing sugar upon acid hydrolysis of dried
cells of P^ chrysogenum Q-176------------------------- 37
16 Fate of the glucose carbon oxidized by resting cells of
P. chrysogenum Qfl76------------------ 40
Ability of various strains of the Pénicillium notatum-
chrysogenum group to assimilate glucose----------
LIST OF FIGURES
Figure Page
1 Rate at which various concentrations of glucose are
oxidized by P. chrysogenum Q-176------- ----- ---- -— — 14a
2 Effect of glucose concentration upon the oxidative
assimilation of glucose for 5 hours by chrysogenum
Q—176— --- --- — ------------------------ — -------- 14a
3 Effect of pH upon the oxidative assimilation of 4xlO”^M
glucose for 5 hours by P^ chrysogenum Q-176------------ 25a
4 Effect of starvation upon the oxidative assimilation of
4xlO“^M glucose for 5 hours by P^ chrysogenum Q-176---- 25a
5 Effect of age upon the oxidative assimilation of 4x10"*%!
glucose for 5 hours by P^ chrysogenum Q-176--------- — 28a
6 Effect of various concentrations of DNP upon the uptake
of oxygen by cells of chrysogenum Q-176 incubated
with 8x10“3m glucose---------------------------------- 28a
7 Effect of 10"% DNP upon the uptake of oxygen by cells of
P. chrysogenum Q-176 incubated with and without
4x10”% glucose --- —---------------------- 29a
8 Effect of 10”% DNP upon the change in dry weight of cells
of P. chrysogenum Q-176 incubated with and without
4xlcF% glucose— -— ----— ---------------------------- 29a
9 Effect of 10”% DNP upon the utilization of 4x10"%
glucose by cells of chrysogenum Q-176--------------- 30a
10 Effect of various concentrations of azide upon the uptake
of oxygen by cells of P^ chrysogenum Q-176 incubated
with 6x10”% glucose----- 30a
11 Effect of 3x10”% azide upon the uptake of oxygen by cells
of P. chrysogenum Q-176 incubated with and without
12 Effect of 3x10”% azide upon the change in dry weight of
cells of PjL chrysogenum incubated with and without
4x10”% glucose— ----- — ------------------ 31b
13 Effect of 3x10"% azide upon the utilization of 4x10”%
glucose by chrysogenum Q-176----------------------- 31b
14 Minced, resting cells of P«_ chrys ogenum Q-176 oxidized
with sodium metaperiodate and stained with sulfite-
decolorized basic fuchsm— ——— — — —— —— — ——— — ————— 34a