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STREAMING ORIENTATION OF AMYLOSE
AND AMYLOSE COMPLEXES
by
•»
Donald Zucker
A D issertation Submitted to the
Graduate Faculty in P artial Fulfillm ent of
The Requirements for the Degree of
DOCTOR OF PHILOSOPHY
Major Subject: Biophysioal Chemistry
Approved:
m Charge of Major Work
Head of Major Department,
(.-■
Dean of Graduate College
Iowa State College
1950
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UMI Number: DP13078
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TABLE OF CONTENTS
Page
INTRODUCTION .............................................................................................. 1
REVIEW OF LITERATURE ........................................................................... 4
Preparation and Properties of Amylose ......................... 4
Separation from amylopectln .................................... 4
Subfractionation of amylose .................................... 5
Structure .............................................................................. 6
Moleoular weight ............................................................ 10
Orientation Theory ................................................................... 14
Rotation of molecules in flow fields .............. 14
Polydlsperse systems ................................................... 22
Birefringence ............................................................................... 24
Theory of double refraotion ................................... 24
Measurement of birefringenoe ................................. 28
Dichroism ........................................................................................ 31
Definition .......................................................................... 31
Theory of dichroism ..................................................... 32
Experimental determinations ................................... 34
APPARATUS ................................................................................................... 38
Meehanloal ...................................................................................... 38
Cylinders ............................................................................. 38
Cylinder drive ................................................................. 38
Polarizer and analyzer coupling .......................... 41
Mounting ............................................................................... 43
Optical ............................................................................................. 43
Light source ...................................................................... 43
Polarizer and analyzer ............................................... 44
Apparatus for visual observation ........................ 44
~ r ? 5 3 ^
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Page
Photoelectric Measuring C ircuit ....................................... 44
Purpose .............................................................................. 44
General description of apparatus ......................... 45
Light so u rce....................................................................... 47
High voltage supply ....................................................... 48
Phototube and pream plifier ....................................... 50
Tuned am plifier and output ....................................... 52
C haracteristics ................................................................ 55
Applications ..................................................................... 59
MATERIALS..................................................................................................... 62
Carbohydrate Samples ................................................................ 62
Reagents ............................................................................................ 64
Commercially available m aterials ......................... 64
Laboratory preparations .............................................. 65
EXPERIMENTAL METHODS AND RESULTS ................................................ 67
Birefringence .......................................................... 67
General technique ............................................................ 67
Preparation of solutions ........................................ 71
Effect of amylose concentration ........................... 74
Potassium hydroxide as in itia l solvent . . . . 77
Effect of contamination with
amyl ope o t i n ......................................................................... 79
Nature of the precipitate developed by
flow ............................................................... 83
Correlation of streaming orientation and
intrinsic viscosity .................................................. 88
Dichroism .................. 88
General technique ........................................................ 88
Preparation of the complex................................... 90
Results on various amylose preparations ... 104
DISCUSSION................................................................................................. 113
Blrefringenoe Results ............................................................. 113
Molecular interaction ................................................. 113
Polydispersity .................................................................. 115
Blrefringenoe vs. intrin sic viscosity ............... 126
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Page
Dichroism Results ................................................................. 126
Molecular interaction ............................................ 126
Polydleperelty ............................................................ 129
R ig id ity .......................................................................... 133
SUMMARY AND CONCLUSIONS ............................................................... 135
LITERATURE CITED ............................................................................... 139
ACKNOWLEDGMENTS ................................................................................. 144
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INTRODUCTION
All starohes, regardless of origin, are alike in
that they are polymers of glucose having 1,4 glucosidic
linkages of the ((-configuration. In nearly a ll starches
there are two kinds of such polymers in varying proportions.
One kind appears to have no linkages exoept the 1,4, being
thus a straight chain oompound. The other variety is
branched, having side ohalns attached by 1,6 linkages. The
firs t of these is oalled amylose; the second, amylopectln.
In this thesis amylose is of primary importance, amylopec
tln being of interest only in so far as it Interferes with
the work on amylose.
Prom the preceding, it is obvious that a ll amyloses
are chemically identical exoept in the degree of polymeri
zation (commonly abbreviated d .p .), that is , in the number
of glucose units per molecule. Considerable work has been
done to determine the d .p .'s, or moleoular weights, of vari
ous amyloses. Both ohemioal and physical methods have been
applied. Chemical methods generally are not subjeot to er
ror due to aggregation, but can be seriously affected by
incomplete reaction and side reactions, degradation of the
sample, and the presence of amylopeotin.
Physical methods Include the application of the
ultracentrifuge, determination of osmotic pressure and
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