Hydrostatic pressure in relation to the synchronous culture of algae

137 Pages·2005·3.58 MB·English

by  Kim, Hubert Lue

HUBERT LUE KIM
B.S .A. University of Toronto, 1951
1 . . A TJn i versity of Ttjr-onto, 1963
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Title of Thesis/~issertation:
Author:
(signature )
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3'0 : LINDA, MICi-GEL, MT310M Ec RITA
A B S T R A C T
Chlorella elllipsoidea cultured using conventional
--
methods under a 14: 10 light-dark regime i n B2ijerir:ck's
medium at 25O +- lo with 700 ft-c illumination resulted ir,
synchronous division w i t h an n number of 4. znder if?cntiezl
conditions of l i g h t intensity, temperature, culture mediun,
and light-dark regime b u t in a closed syskem which was 2esiqnc3
t o study thc effects of hydrastatic pressure on syncl-lrcnous
algae cu? t.ures, synchronous cell divisi-on did nat o c c ~ ~a~r. ii
the n nunbcr was 2 or less a t 1 atn. The xost signific::~t
difference i n the culture conclitions criszs b c c a v c t thz cc,.-
r e l a t i v e l y constant throughout the growth cycle i n thz open
system while the gases vary i n concentrdtion contirico1:~sJ.y
during the cycle i n the closcd systen. In the closrd systcz
period; the reverse is true i n the dark. J.;'.nal:;st?a of the
of hydroc +;aijc . pressure on Che physiol oc;y 2nd morp3oloyy of
successful culturz i n a closcd system. Various niodifica'iior!~
of cul'icr-2 conditi.ons w e r e made in 3tt.empts (on1.y partially
succc~~sful.t)o obtain equal growt5 i n the two s y s t e n ~ , .
The effscts of variations i n some cnvironrnmtal parameters or.
t h e n number, dry weight, protein concentration, and c e l l size
i n the open and closed systems were investigated. With varicc!
growing conditions of light intensity (15-706 ft--c), liqht-
-
dark regimes (14: 10, 16: 8, 18: 6, 6: 18, 8: 16, 10: 1.1) ,
media (Ezij erinck, Burr, Sorokin, Tamiya and Morinura, ar.d
Beijerinck's modified with bicarbonate), C02 concentration
(0-100% saturation), and the addition of organic substznces
(glycolate, ascorba-te, dithiothreitol) the n numher and the
r a t i o s of increase i.n the other growth c r i t e r i a were 4 in the
open systcm but never more than 2 i n the cl.osed system.
The n number and ratios of increase i n dry wcigl:~,
protein content and cell size were 4 i n the open sl-stem zt
atmospheric O2 concentration but only 2 or less a t high O2
(50 and 95%) concentration. These results indicate that hiqh
O2 is probably responsible for the descrepancies i n the n
number and the r a t i o s of increase of the other c r i t e r i a beJi~ccil
the o p c ~an d closed systems. Photosynthetically evclvec!
O2
i n the closed system during the light period a2pcars t o be
.
sufficient t o induce thi s Warburg Effect
It was hypothcsizcd that the Warburs Effect i s
responsible for the observed differences i n ificreases in the
growth c r i ~ t c r i ai n the open and closed sys tcius. E:.:p~l-i~.ents
were carried out t o determine i f photosynti1c:licci11;, e ~ o l ~ ~ - c i l
O2 was inhibiting cell growth. Using the Eiolagical Osyqcn
Monitor algal. samples were exposed t o O2 concentrations
varying from 0 Co 100% saturation a t 25O with 700 ft-c
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illumination. On the basis of these results which have
indicated a decrease in the relative rates of O2 evolution
at high 02 concentrations, it was concluded that O2 evolved
in the cl2scd system could be responsible for the inhibition
of photosynthesis.
Synchronous growth under pressure has not yet been
obtained. However, pressure effects on cell divisicn were
investigated. The n numSer was 4 in cells dividicg \:bile
exposed to pressures of up to 200 atin, but pressures above
335 atm compl~telyi nhibited cell division. A possible
mutagenic cficct of pressure csposu1.e 011 zlqal celis was
considered.
A C K N O W L E D G E M E N T
I wish t o e::press my sincere appreciation t o
Professor William Vidaver f o r i n i t i a l l y suggesting the
research problem. Also, f o r h i s patierit and unceasing
advice, guidance and freely given encouraqe;nent throughout
t h e research and preparation of t h i s thesis. Many thanks
are a l s o gi~:,-n t o Professor Vidaver f o r designing the
pressur? vessels and to Mr. Frank Wick and h i s staff of
t h e Sinlcn Fr~:~,eU.in i~icrstjy Machj.ne Shop f o r th2i.r excellent
work i n t h e construction of t h e pressure vessels.
I aw a l s o grateful t o Dr. H. Speer and D r . L. Druekl,
members of my cjraduate committee, for t h e i r hel-pful suga,estior:s
and criticisms. Many thanks go t o t h e other m a h e r s of t h e
Department of Biol.ogica1 Sciences who have helped i n several
ways to make t h i s work possible. I owe a great deal of
gratitude t o the late Head of the Dcpartwent, Professsr
C. D. Nelson, f o r h i s valuah1.c assistance an3 f o r the ~ 3 n y
fringe benefits accorded t o me a s a graduate student.
My thanks go t o Nr. Gordon Ashby for 11i.s excellent
drawings, for h i s s k i l l f u l technical assistance and helpful
surjgestio;ls; to the stazf of thc Department and my colleaques
who have made t h i s work possible. I wish t o sinccrell- thank
Mrs. Daphnc Chin for typing the thesis. Many thanks go t o
Mr. Ming Won9 for h i s valuable assistance.
To my wife Linda, I owe much for her help and
encouragement throughout this study. Her continuous assis-
tance and understanding were essential t o the con12letion
of t h i s research.
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T A B L E O F C O N T E N T S
Page
................
ACKNOWLZDGEMENT,.................,...... iv
..............................
TABLE OF COhTENTS......... vi
...........................,...........
LIST OF FIGUXES.. x
I. Metabolic Processes Ai-~d
..................
Enzymatic Actjvit~es 9
111. Son12 General Effects Uf I ~ s s s ~ ; l r e
...........,......
On Macromolecul.cs...
.............................
IV. Iblutations 18
. . .. . . . . . . . . . . . . . . . .
CHAPTER 2 -- i4ATERfAJS AND >IET!SODS.. 21
..................
I. Synchronou Cultures 2 1
. . . . .. . . ... . ..
11. Pressure Vesst.1 Systm~.. 2 0
.............
111. Biological Oxygcn Xonitcr 4C
.
. . . .
I. Variatj ons In Groiv.t:l; Parmctcrs.. 44
. . . . . . . . . . . .. . . . . . . . . . . . . . . . .
1. Ned j-a 44
2 . Gases............................. 44
...........,.....
3. Light-dark Rcgjm? 47
...................
4. Liqht Intcnsjty 4 E!
. . . . . . . . . .
5. Nitrztc-cnr ic:hcd Fndi urn. 53
.
G Glycol.c .? t..e. ..S.l..ip.i>.;.er.n.c!.l ..Le.d. ...........
Medi.uni 5 5