Antibiotic substances from yeasts

47 Pages·02.163 MB·English

by  Nakamura, Mitsuru

ANTIBIOTIC SUBSTANCES PROM YEASTS
A Thesis
Presented to
the Faculty of the Department of B acteriology
U niversity of Southern C alifo rn ia
In P a rtia l F u lfillm en t
of the Requirement fo r the Degree of
M aster of Science
by
M itsuru Nakamura
June 1950
UMI Number: EP55015
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This thesis, written by
MITSURU ..NAKAMURA
under the guidance of h%3.... Faculty Committee,
and approved by all its members, has been
presented to and accepted by the Council on
Graduate Study and Research in partial fulfill­
ment of the requirements for the degree of
MASTER OF SCIENCE
Date 1M M ..1950
Faculty Committee
TABLE OF CONTENTS
CHAPTER PAGE
I . THE PROBLEM ......................................... 1
Statem ent of the p r o b le m ............................... 1
Importance of the s t u d y .................................... 1
I I . REVIEW OF THE LITERATURE.......................... 3
I I I . METHODS AND RESULTS............................... 15
EXPERIMENTS................................................ 15
Screening of y easts fo r a n tib io tic a c ti­
v ity by ad ju stin g thep H ............................. 16
Best carbon source fo r a n tib io tic pro­
duction by Torula u t il i s . . . . . . 19
Best nitrogen source fo r a n tib io tic
production by T. u t i l i s .............................. 20
E ffect of shaking the cu ltu re on a n tib i­
o tic production .................................... 22
E x tractio n of the a n tib io tic substance
from the m edium ........................................ 23
Use of sy n th etic media . . . . . . . . 26
O btaining a b a c te ria l spectrum fo r the
a n tib io tic . . . . . 31
T esting fo r heat s ta b ility of the
a n tib io tic . . . . . 31
Toxicity te s ts on r a t s .................................... 34
H i
CHAPTER PAGE
XV. SUMMARY AND CONCLUSIONS' . . . . . . . . . . . . 35
BIBLIOGRAPHY ............................................................................................. 38
LIST OP TABLES
TABLE PAGE
I. The Composition of U niversal y east Medium*. . 15
II. Yeasts Screened fo r A n tib io tic A ctiv ity . . . 15
I I I . A n tib io tic A ctiv ity of F iltra te s of Y easts
Grown a t Varying PH Levels A gainst S. aureus 17
IV. A n tib io tic A ctiv ity of F iltra te s of Y easts
Grown a t Varying PH Levels A gainst E. o o li 17
V. Further Studies on the E ffect of PH on the
Production of A n tib a c te ria l Substances . . 18
VI. E ffect of Carbon Source on A n tib io tic
Production by Torula u t il i s . . . . . . . 20
V II. E ffect of N itrogen Source on A n tib io tic
Production by Torula u t i l i s . ..................... 22
V III. Measurement of In h ib itio n Produced by E ther
E x tract of T. u t il i s medium . . . . . . . 24
IX. Compositions of S ynthetic Media Used fo r
A n tib io tic Production • • • • • • • • • • 27
X. End-Products of Yeast Ferm entation of Sugar in
R elative Proportions U sually Present . . . 50
XI. P ro p erties of E n d -P ro d u c ts ...................................... . 50
X II. B a cterial Spectrum Obtained from an A n tib io tic
Produced by T. u t il i s on S ynthetic Medium 52
CHAPTER I
THE PROBLEM
A n tib io tics are playing an im portant ro le in the fie ld
of m edical sciences today. Many a n tib io tic s have been is o ­
la te d and described from b a c te ria , molds, and streptom yces,
but very l i t t l e work has been reported on the p o s s ib ility of
producing an a n tib io tic substance from y e a sts.
Statem ent of the problem , The purpose of th is study
was to survey ten yeasts fo r the production of an a n tib io tic
p rin c ip le . If a y east was found th a t formed an a n tib a c te ria l
agent, the aim was to determ ine the optimum conditions fo r
maximum a n tib io tic production, and to is o la te the agent so
th a t d e ta ile d stu d ies of i t s spectrum could be obtained.
Importance of the stu d y . D espite the fa c t th a t many
a n tib io tic s have been discovered, re la tiv e ly few are used in
m edicine. Unless the newly discovered agent dem onstrates a
m arkedly d iffe re n t spectrum again st pathogens i t is not
fe a sib le to develop i t com m ercially in com petition w ith
compounds fo r which the fac to rs of production are known.
The fa c to rs c o n tro llin g the production of y easts are
more fu lly understood than fo r any o th er group of organism s.
A good a n tib io tic from th is source could be produced cheaply
and in large q u a n titie s. I t was hoped th a t in searching fo r
2
a new a n tib io tic fronr y east, one m ight be* found th a t would
be e ffe c tiv e ag ain st a wide v a rie ty of organism s.
CHAPTER II
REVIEW OP THE LITERATURE
For many years there has heen a popular theory th a t
abscesses and pimples w ill disappear when y east is ingested
and th a t i t was of im portance also in the treatm ent of other
skin d iseases. Long before s c ie n tis ts re a liz e d th a t y east
contained vitam ins, physicians were using i t fo r therapy
ag ain st skin d iso rd e rs.
The a n tib a c te ria l a c tiv ity of the y east was f i r s t
rep o rted by Sergent (1903), who rep o rted a substance p ro ­
duced by yeast which was toxic to Staphylococcus.aureu s.
B urnett (1905) reported the use of yeast as a d isin fe c ta n t.
Hayduck (1909) m entions in h is paper th a t Lange, Henneberg,
and Hayduck have d etected the presence in c e rta in cereals
of substances th a t are poisonous to yeast when dissolved
to g eth er w ith sugar in d is tille d w ater. He was able to
o b tain a sim ila r substance from d i s t i l l e r ’s y east when i t
was dried rap id ly a t high tem peratures, and ex tracted w ith
w ater containing 0.1 per cent hydrochloric acid . He found
th a t no such poisonous e x tra c t is obtainable from yeast
th a t has been dried slow ly, owing probably to the actio n of
try p tic enzymes upon the poisonous p ro te in . He ex tracted a t
both 25° C. and a t 50° G., but found th a t an e x tra c tio n a t
50° C. gave b e tte r re s u lts . At 50° C., the e x tra c tio n is
4
completed in two hours. His experim ents in d icated th a t n e ith e r
su lfu ric acid nor a c e tic acid can replace hydrochloric acid
in th is e x tra c tio n process. To prepare a poisonous e x tra c t
from brewer*s y east, he found th a t i t was necessary to remove
some of i ts lime by d ilu te acid previous to the e x tra c tio n
proper. Calcium s a lts prevent the poisonous e ffe c t. If the
y east was weakened by storage before being d ried , a le ss
poisonous e x tra c t re su lte d . The a ctio n of th is so lu tio n was
in te n si fied by d ia ly s is .
L ater in the same year, Fernbaeh (1909), using approxi­
m ately the same procedure as Hayduck (1909), found an activ e
substance th a t could k i ll b a c te ria . He ex tracted a m acera­
tio n of d essicated y east w ith d ilu te (1:1000) hydrochloric
acid fo r 19 hours a t 37° C ., and f ilte r e d the so lu tio n . A fter
n e u tra liz a tio n , the f i l t r a t e exh ib ited marked to x ic ity to the
yeast i ts e l f as w ell as to various o th er stra in s and to
b a c te ria . In h is experim ents he used one Bacterium c o ll and
one Staphylococcus aureus pyogenes s tra in . The so lu tio n
proved to be b a c te ric id a l to both of these, though somewhat
more inten se ag ain st the la tte r . He found the toxin to be
v o la tile . Upon d is tillin g the f i l t r a t e under dim inished
pressure such th a t the tem perature did not ris e above 40° C.,
the to x ic ity was tra n sfe rre d to the d is tilla te w hile the
re sid u a l liq u id was in a c tiv e . No reactio n s were obtained fo r
aldehydes—notably form aldehyde--to which b a c te ric id a l actio n