Of NATURAl CES II ENGINEERING · turgut bmh

AN ANNUAL PU L1CATION VOLU f 9 JUN 1980

HAC EIIEPE IUllEIlM Of

NATURAl CES II ENGINEERING

A 8U LLETIN PUBLISHED BY FACULTY OF SCIENCES OF HACETTEPE UNIVERSITY

HACETTEPE BUllETIN OF

NATURAL SCIENCES II ENGINEERING AN ANNUAL PUBLICATION VOLUME 9JUNE 1980

EDITOR I TURGUT BMHltAN

EDITORIAL BOARD (HACETTEPE BULLETIN OF NATURAL SCIENCSS AND ENGINEERlXC)

TURGUT BAeKAN (CHAIRMAN OF EDITORIAL BOARD)

BUNA BOZCUK SOLEYMAN GONAY

MANAGING EDITOR ART DIRECTOR I FAHRETTtN SAvelA

PUBLISHED BY THE FACULTY OF SCIENCES OF HACETTEPE UNIVERSITY BeytepeAnkaraTurkey

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Printed by the Faculty Press 1980

VOLUME 9JUNE 1980

HACErrEPE BULLETIN OF

ANDNATURAL SCIENCES ENGINEERING CONTENTS 1 Control of the Cellulase Synhesis in

Cellulomonas Flavigena (Selulaz Sentezinin Kontrolu)

Nazif Kolankaya

11 Grouping some Variables of the New-Born Baby BOIS (Yeni Dogmu7 Erkek 90cuklara Ait Baz~

Degi~kenlerin Grupland~r~lmas~)

Soner Gonen

21 The Use of Prior Distribution in the Design Criterion for ~arameter Estimation (Parame~e Tahmini t~in Duzenleme ~l~ushytunde ~nsel Dag~l~m~n Kullan~m~)

suleyman Gunay

27 Prolon gemen t d une Semi -metr ique (Bir Yar~ Metrigin Geni~letilmesi)

Ahmet Abdik

31 Generalized Distance Function and QuasishyUn iform Spaces (Genelle~tirilmi~ Uzakl~k Fonksiyonlar~

ve Simetrisiz Duzgun Yap~lar)

Yus uf Ayd~n

37A Note on Abels Theorem for Open Riemann SurEaces (A~~k Riemann Yuzeyleri t~in Abel Teoremi)

Turgut Ba~kan

47 Some Relations Between Behavior Spaces (Belirtme uzaylar~ Aras~nda Baz~ Bag~nshy

t~lar)

Turgut Ba~kan

CONTENTS (Cant)

57Remarque sur lBgalite des Fonctions Zeta des Corps Globauyen (Global Cisimlerde Zeta Fonksiyonlar~n~n E~itligi Uzerine)

Mehpare Bilhan

bull j650n Invariant Vector Measures

Uzerine) De~erli Degi~iiZ tJ1~umler

Dogan 90ker ttlmiddotmiddotmiddotmiddot bullbull

710n Vector Property

Valued Integrals With the Darboux

(Darboux ~zellikli Vektor Degerli tntegshyrailer Uzerine)

KAz~m Guner

79 Applications of Generalized Behavior Spaces to Conformal Mappings (Genelle~tirilmi~ Belirtme Uzaylar~n~n

Konform Donu~umlere Uygulanmas~)

cosk un 1ayfur

89L-Serine Dehydratase Synthesis By Pseushydomonas Aeruginosa (Pseudomonas Aeruginosada L-Serin Deshyhidrataz Sentezi)

Atilla Atalay

97 A New F~sh Species From Lake Van (Cypshyrinidae) (Description) (Van Golunde Bulunan Yeni Bir Bal~k Turu (cyprinidae) (Tur Tan~m~)

Mustafa Kuru

l03Key To the Inland Water Fishes of Turkey (PartI) (Turkiye t~ Sular~nda Ya~ayan Bal~klar~n

Tan~ Anahtar~) K~s~mI

Mustafa Kuru

CONTENTS (Cant)

113Key to the Inland Water F~shes of Turkey Part II (Turkiye t~ Sular~nda YasectaYf Bal~klar~n

tmiddot ~n~ Apa~~ar~) K~s~mII I Mustafa Kuru j

123Key to the Inland Water F~shes of Turkey Part III Cyprinidae (lurkJlJe t~ Sular~nda Yasectayan Bal~klar~n

~h~~~stitar~ K~s~m III Cyprinidae

Mustafa Kuru

r

HACETTEPE BULLETIN OF

NATURAL SCIENCES AND ENGINEERING VOLUME 9 JUNE 1980

CONTROL OF THE CELLULASE Sn1HES1S tN CELLULOMONAS FLAVIGENA

SELULAZ SENTEZNN KONTROLU)

NKolankayaX

SUMMARY

In this study effect of some external factors which control the cellulase enzyme synthesis in Cellushylomnas flavigena A~CC 482 was investigated The synshythesis of enzyme was found to be repressed by readilymetabolized carben sources added into Cflavigena culshytures grewing en cellulose In add~~ion tc the carbon sources the concentrations of dissolved oxygen and yeast-extract in culture media were also effective on cell~~ase fermation 24-DNP at final concentration of 5xlO M in culture medium increased differantial rate ef the enzyme synthesis The existence of an inverse relatienship between growth rate and enzyme productionof Cflavigen was a comman observation af the study

INTRODUCTION

Synthesis ef cellulases by fungi and bacteria is considered te be induced by cellulose substrates Howshyever since cellulose is high-molecular weight composhyund its penatration inte cell is problematical Iccorshyding to the cncept of JACOB and MONOD(1961) inducer is an exegeneus substance that penatrates into cell and derepresses the synthesis of enzyme protein The data obtained in studies dealing with cellulase formation suggests that there was inverse relationship between metabolic rate and cellulase production of bacteria

and fungi(YAMANE etal1970HULMB and STHANKS 1971)

2

In present work control ef the cellulase syntheshy

sis in Cflavigena cultures was studied

X Hacettepe University Science Faculty Departmentof General Biology Ankara Turkey

MJTERIALS AND METHODS

I Organism and Culture Conditions

Cellulemonas flavigena ATCC 482 was provided from American Type Culture COllectien USA Steck cultushyres were maintained in slants made of stock-culture

Agar(Difce) Working basal medium was consisting of O 1K~40O~CI005MgSO~ and OO~ yeast-extract(Dir8o) Inte the basal med1llm water-soluble derivati shyve er cellulese CMC(Carboxymethylcellulese) and otshyher carben sources were added after they were sterili shyzedseparatelyErlenmeyer flsks containing 50 or 100 ml er sterile culture medium were ineculated with bacteshyrial suspension prepeared by suspending the bacterial cells in slant tubes with 10 ml of sterile saline soshylution at ratio ef 01 ml suspensien per 100 ml ef medium Culturatiens were carried out with Pgycretheshyrm Incubator Shaker(New Brunswick Co) at 30 C with dirferent peried ef growth time Disselved oxygen conshycentratioDs in culture flasks centaining 50 ml ef meshydium were measured with the aid er Oxygen electrode(YSI odel) fer each 8hakinlaquo 8peods(100-300 rpm)

lad the rate or Oxygen tran8fer into mediua was reushynd as suggested by ARNOLD and STEELE(1958)

IIleurbullbull of Grethbullbull~O1UltfS Activitr

The grewth or Crlavigena was measured by reading eptical deasity et culture fluids at 600 nm by meaDS er spectreaio 20 model ef Besch aad Leb spectrophetoshymeter

Culture supernatants were used as enzyme seurce in enzyme assays The method described by NISIZAWA et al(197l) was used fer measuring the cellulase activishyty as CMCase In experiments a unit CMCase activity was described as the ameunt ef enzyme preducing ~g reduciag sugar in a iAute at the experimental cendishytiens The ameunts er reducing sugars i enzyme and incubatien selutions were measured by DNS(Dinitresali shycilic acid) precedure(MILLER1959) and- calculated as glucese en standard curve prepeared with glucose Sinshyce the enzyme synthesis is occured during exponantioshy

3

I

I

nal growth phase of Cflavigena the differanta1 rashytes of enzyme synthesis were found usinr the equationoffered by WELKER and CAMPBELL(1963) for amylase proshyducing organism as follows

RES1LTS

IEffect of Carbon Sources on Cellulase Synthesis

When the growth and cellulase synthesis in Cf1ashyvigena were considered itwas observed that the orgashynism produced the enzyme throughout logarithmic phaseof growth in presence of 10 CMC as carbon source(Fig 1) Reducing sugar in the culture was also increased during the logarithmic growth phase The CMCase formashytion in terms of differantial rates has been effected by CMC concentrations Synthesis of the enzyme was inshycreased up to the O~ CMC and then slightly decreased (Fig2)While the addition of glucose and cellobiose were activating the groth in cultures grown on CHC these two sugars were both repressing the cellulase formation (Fig 3 and 4)Same phenomenon was observed with the other carbonsources which were added into the culture media at 24 hr of culturation(Table 1)The most activily repressing carbon sources were of that on which Cflavigena was shown to grow with hi~shyher growth rates

IIEffect of Physiological Conditions on Cellulase

Growth of Cflavigena was found to be proportionshyal to the concentration of dissolved oxygen in culture medium(Fig5)But the formation of cellulase was neshygatively correlated wit the oxygen concentration Amoshyunt of yeast-extract in cplture medium has also affected growth and cellulase formation in culturesThshyere was an inrease in the growth of organism doe to the increase in amount of yeast extract concentration up to 020 in the medium In contrast to increase in growth enzyme formation in culture was decreased (Fag 6) 24 DNP has influenced the enzyme synthesis and growth of the organism inversly(T~ble 2)Presence of 24 DNP in culture medium at 5xlO-M concentration caused decshyrease in growth but the increase in CMCase synthesisof Cflavigena

4

TABLE IEffect of various carbon sources on synthesis of cellulase in Cf1avigena cultures

Carbon Sourcex CMCase Growth Rate xx

(AU~0D600nm)

cMC(Control) 2132 8 Lactose 853 18 Na-acetate 725 20 Mannose 720 21 Glycerol 720 25 Fructose 450 50 Ga1actese 265 60

X Carbon seurces were added into media at 24hr ef culturation to give final concentration of 002 Before the addition of carbon sources 05 CMC was

sole source of carbon in media

XX Growth rates for each carbon sources were found by grawing Cflavigena in medium containing re1avant carben seurce at 01 concentration

I I I I I I

--ll--X f

01- - 120

~o--o-o 0 I- oE I

600 cOmiddot6 f- - 100 u 0 0

~

0

01 E u0 ~o--~400 ~ 04 f- - eo t -- ----

E e ~

u Ill ---

vi ~ _ rio ~

200 0middot2 - 40- -~-- x- 1

0 I I I I I

20 30 40 SO 60 70

Growth PHiod (hrJ

FigIGrowth and Cellulase Formation in Cflavigena Or~anism was ~rown in basal-medium containing 10 CMC XCMCase activity (IVml)oGrowth (0D600 )Reducing sugar- ()agml) nm

5

Ii I iI I

-E c o g 50 o o ltJ-J ltJ 40 --U lt

~

~ 30 x j

u ~ u

01 02 0 06 08 10 CMC

Fig2Differantial rates of CMCase synthesis in the presence of var~ous concentration ofoCMC in medishyumCells wer~~rown for 24 hrs at 30 C 150rpm

020

016 o

40

30

20

10

~ ----------

o ~

E CI c o ~ 012 ~

)

~

oC( u Z u

o

_ Eltract (difcD )middotf hilt

Fig6Effect of yeast-extract on the growth and rate of CMease synthesis of Cflavigena oGrowth of organism Rate of CMease synthesis Culturatishyons were cal)ried out in presence of 05roCMC for 24 hr at 30 C

6

CMC as e Act (~U~00600nml s- QI C

~ --~

~ - ~ 0 0 o n bull 0

11

L

s-

-0 7

o bull

J J I I

o I o bull I II

bull 0 o bulll7gt o

I I I o 0 0 0 N C7 ~

o 0 0 0

Growth (00 600nm 1

Fig3Effect of plllcose on the growth and rate of CMCshyase synthesis of Cflavipena 002 glucose was added into culture growing on 05 CMC at 10hr of culturation 0 growth of controL~)Growth of control plus glucose 0 Rate of CMCase synthesis of contro~---Rate of CMCase synthesis ef centrol plus glucose

7

----- -shy- -shy~- _- - 0--- --0

0--- 0

I I 0

I I 1

1

+ -E20

c Cgt Cgt o o ~ 16 gt ltI

( 12 u 2 8 u

4

20 30 40 so (rowlh Penod lhr )

10

-030 E

c g g

o o

020 j e

010

Effect ef cellobiose on the growth and rate ofFir-4 OMOBae synthesis ef Oflavigena 002 cellobishyose was added inte culture growing on O5CMO at 10hr of culturatioB ---oGrowth of control--~Growth of control pshylus cellobiose---oRate of OMOase synthesis of control~Rate of CMOase synthesis of control plus cellebiose

8

020

018

~

E c c c

0016 o

~ 014 ~

012

010

o X o

gt - 4~

100 1~ 0 200 2~0 300

S~aking rale ( r p ml

60

~o ~O

~40 N~ 40 E

~ c 0

0 -s 0

30 3D 0

s J ltI-JN ~

0 C( 20 20

~ Ir U

10 10 u 20

Fig5Effect of dissolved o concentrations on the growth and CMCaae syn~hesis of CflavigenaXO transfer rate into medium oGrowth of organism ARate of CMCase synthesis Culturation was ca5rshyied out in presence of 05CMC for 24 hr at 30 C

Table 2 Effect of 24 DNP on thexgrowth and cellulase synthesis of Cflavigena bull

24DNP Growth CMCaae

(1101) (0Dmiddot 600nm) ( [1 0D60nm)

000 (Control) 016 2120

5xlO-5 012 3160

X Culturations were uBrried out in presence of 05 CMC for 48 hr at 30 C

9

DISCUSSION

On the basis of our present data limitation in metabolism of the Cflavigena is resulted in induction of cellulase synthesis It is apparent that the cellushylase synthesis in Cflavigena cells is more effectiveshyly repressed by the easily assimilated carbon sources (Fig34 and Table 1) Induction of some depolymerases in culture of bacteria and fungi was shown to be posshysible when the growth of organisms was slowed down by limiting the consumption of carbon sources(HSU andVAshyU~HN 1969 LABONAK and PAVLOVSKAYA1975 HULME and STRANKS 1971) Comparativ-ely recently initiation of the cellulase synthesis by lactose in Trichoderma li~shynorum was regarded as a result of slow consumption 0 sugar by the organism (LABONAK and PAVLOVSKAYA 1978) Induction of cellulase formation of Cflavigena by celshylulase is probably dependent on slow degradation of cellulose by the enzyme already present in small amoushynts in the cell Therefore cellulase may be semi-conshysitutive in Cflavigena as in Pseudomonas fluorescens (YAMANE etal1978)

In experiments the cellulase formation of Cflashyvigena was enhanced due to dissolved 0 and yeast exshytract concentrations wnich are not optfmal for growth (Fig56) Restriction in ATP bynthesis and on growth of Cflavigena was also resulted in increase in cellushylase formation(Table 2) As proposed by HULME and STRshyANKS 1970) for celluloytic microorganisms limitation in metabolismof Cflavigena may be the main mechanism for its cellulase synthesis

Manuscript Received in April 1980 REFlRENCES

1 ARNOLDBH and STEELER1958 Oxygen supply and demand in aerobic fermantations In Biochemical Engineering EdRSteele New York McMillan

2 HSUEJ and VAUGHNRH1969 Producticnand catashybolite repression of the consititutive polygalactshyronic acid tran-eliminase of Aeromonas liguifacienshyce JBacteriol~8172-181

3HJLMEMA and ~ANKSDW1970 Induction and the regulation of production of cellulase by fungiNature 226469-470

4H~JLMEMA and STRANKSDW1971 Regulation of celshylulase preduction by Myrothecium verruceria grow on non-cellulosic substrates J Gen Microbiol bullsect2 11-5shy155

J

10

5 JACOBF and MONODG196l Genetic reGulatorymechanism in the synthesis of proteinsJ MolBioLl 518

6 LABONAKAG and PAVIJOVSKAYAZhI1975 Derepressshyion of cellulase synthesis in Trichoderma ligno~lm during limited conaumpt Lon of readly assimilated cashyrbon sources Mi crobiology(Trans Russ i an ) 44 25-28

7 LABONAK AG and PAVLOVSKAYAZhI1978 CoI1Gti tu t igt ve cellulase synthesis in Trichoderma lignorumMicrobiology(TransRussian) 46341-45

8 MILLERGL1959 Use of dinitro salicilic acid reashygent for determination of reducing sugar AnalChemmiddot21426-428

9 NISIZAWATSJZllKIHNAKAYAMAM and NSIZAWAK 1971 Inductive formation of cellulase by sophorose in Trichoderma virideJBiochem70375-385

10WELKERNE and CAMPBELLLLI963Induced biosynshythesis ofoC-amylase by growing cultures of Bacillus stearothermo~hlis JBacteriol 61196-1201

11YAMANEK S~UKIHHIROTINIM OZAWAH and NISIZAWAK 1970Effect of nature and supply of carbon sources on cellulase formation in fseudomonas fluoreijcens yar cellulos JBiochem sectZ9-l8

OZET Bu ~a11qmada Cellulomonas flavigena ATCC482 susectu

lizerinde sellilaz sentezini kontrol eden d1q etkenler araqt1r1lmaya ~al1q1ld1 Selliloz var11~1ndan liretilen CflaviBena kliltiirlerinin ureme ortam1na eklenecek koshylay metabolize edilen karbon kaynaklar1n1n sellilaz senshytezini bask1lad1klar1 saptand1 Eklenen karbon kaynakshylar1 d1~1nda sellilaz sentezi uzerinde ortamdaki oksishyjen ve maya-ekstresi konsantrasyonlar1 de etkin olma~4 ta idi Ayr1ca 2-4-DNPnin kliltlir ortamlar1nda 5xlO konsantrasyonunda bulunmas1 durumunda enzim sentezinin differasiyel h1z1n1 artt1rd1~1 ~ozlendi Genel olarak Cflavigena kultlirlerinde organizman1n ureme h1Z1 ile enzim uretimi araS1nda ters bir iliqkinin var oldu~u saptand1

89

L-Serine Dehytlratarc Syntlcis ny

Pncudomonas aeruqdnos a

P3etlCO~10nas 2eru0inosu da

L-Serin f)ehirlratu~ scntezi

Atilla itu1lti

sm ~fS

L-scrine dehyCratase (L-serine hylrltrlyase

EC ~bull 2ll3proluction was stucliecl in Pseudomonas

aeruginos a Qfv~D-lil68 Imiddotla=dnwn L-serine dehydratltl3e

activily Has obtained when 05 ~ nI Cl was supplied1an nitrogen source in the basal rtediu~l containing 1 ~~

0cef e~dracO3 ~~ I IIP0 o lncl 001 ~~ IJS047 E2 20 Variou carbon nources inhibitc(l or c~id not ef~ect the

onzyiio ~oration

Il~ROTYJCIOI~

REG7l~ and co-xorkers (1~69) showed that chronic

granulocytic Leukemia as middottell as normal human ilarrOl

ccll~ cpryarcntly reluire Derinc for gr~nh ~~i3

finc1inCj ~Ult]lests that L-nerinc (~chyrrCltasc (LS)-1igh

~1facctcJe Univerni ty Science -acultyIolecular riology JlcClrtilcnt reytec Cltl-~rnlsnJra ur~cy

~

90 I i

1e 11-efu11 Ln 1C)~j2 cgter2~1Y I I

or li )11rOe iELS)l C ill (1)73) middotc-ccne(~ 117 )ltlcJ-cC~21 middot~raLle n OllJ[ thot Chror~O)2ct~rill~

rJJshy- 11 (1)7S) - 1Jc) creencd it roric of

In te ~reen t~v 118 nutritional factors affecting the enzyme formation were studied

CULlrtcr1il~tcr re2-c~J 11 cVclolent ~enJccr lSArray

~Cltic_r~2gthc cJ11l~ F -aintClinc( on a -1e~i1u

contaLninr-] Jra ~)er Li 0) Pegt~on 10 rcc ertct C

I r

Ihourc at 37~C on C roi~y 8hlgter(1ml Ir11iJic SciCo)

1lt 17- rCNrinCel1 Jere )1Clrve~tc y ccncriuglt11io1 I (()O~0 or 10 li1) m rCGupcnlel i-t r 11 of Hater I

91

IIIEnzyme ansay

The ~~eto acid produced by the enzyme was determined

)y the direct rne th od of FRIEDEl-iANN and IIAUGEt (191middot3)

A 01 ml of cell suspension -1a mi~~ed Hith 09 ml of

(A) buffer (01 l-i bdratepII RS 001 r1 HgC12 and 01

r L-serine) and incubated in a 10 X ) ( iU11 tube with

-eciprocal EihaJing in a water ~)ath at 37middotCDuplicate

s arrpla tubes were incubated Hi th a control tube in (A)

~)1Jffer ithout L-serineAfter 10 -lin the reaction was

sto~pcd by adding 1 ~l of 1 N HCl and 03 ~l of 01 24-dinitr09henylhyorazine in 2 n RClThe mixture was

further incubatert for 10 min at 37middotCancl then 2 ml of

2 n NaOH wan added The absorbance of the solution was

dete~ined at 417 rum in a Coleman spectrophotometer

The a~ount of pyruvate liberated Has calculated by

reference to a standard curve prepared with crystalline

sodium pyruvate

A unit of L-serine dehydratase waG defined as the

amount of enzyme catalyzing the fozraa tLon of 1 11 of

~yruvate per min under the ooridd tions describect aJove

The remainder of the cell suspention was dried

(overnight at lOOmiddotC) and the dry weight dete~nined

RESULTS A~TD DISCUSSION

Different nitrogen sources were added to the basal

medium containing 1 Beef etract03 ~G andEZHP04 001 r-1gS017 H20 to examine the effect on the enzyme

formationAmong the nitrogen sources tested only llli~Cl

increased the enzyme activity (Table 1)

The other nitrogen sources increasedlhe grorth

rate of culture but inhibited the cnzyne formation The

effect of l~I~Cl on the yield of LSD was further 3tudied

92

syntheGi3 from 0081 to 0221 Irymg dry weight(Table~

T~)le 1Effect of nitrogen sources on the formation of L-serine dehydrataseX

litrogen source

Control Yeaci Etruct rYPlonc Casein Hyckolysate Peptone Corn Steep Liquor NH1-0l -

UTserin dehydratase Dry wt of IUmg dry wt of cells cells rlgml

0085 0011 0006 0003 0015 0005 OlSO

098 182 190 107 151 123 056

~he mecliwn contained 1 ~~ beef e=tractO3 ~~ K2IIP04 anl O 01 ~~ r~gSO4 7 H20

Table 2Effect of a~oniun chloride concentration on the formation of LTserine dehydratasex

L-serine dehydratase Dry wt of cells IUmg dry wt of cells mgrl1

o 01 05 10 15 20

00801shy0083 0221 0058 0055 0074

098 098 077 0ry8 060 OSmiddot

The basal medium was the same as in Table 1

Addition of 05 mI~Cl increased the enzyme synthesis

from 0084 to 0221 IUmg dry weight (Table 2)

In their previous studies PARDEE and PRESTIDGE (1955)

found the highest activity of ~serine deaminase when

they used Casein hydrolysate and yeast extract as a

nitrogen source in EcoliIn addition ISENBERG and

NEHNAN (1974) ahowed the inhibition of enzyme formation

in Ecoli K 12 with the addition of ammonium sulphate

to the uinima1 medium containing pr01ineleucine and

93

glycine These finding suggest that the Pseudomonas

enzyme iay be different from that of Ecoli

rwn amino acids were added to basal medium

together with 05 lili4

Cl gr owt h rates of cultures were

increased 0ut the enzyme forunation either inhibited or

unaffected (Table 3)

HOHeverprevious studies of PARDEE and PRESTIDGE

(1955) and ISEhBERG and NEmIAE (197~) have reported

LSD from Ccoli induced by glycine and leucineThe

effect of carbohydrates and some carboxylic acids as

a carbon source on the production of LSD was middottested in

a hasal rnecHurn con taining 1 ~~ Pepton 0 bull 3 ~~ I~ 2IlPO4 and

001 lmiddotgSO-17 H 20 It was found Uwt carbon sources

with the exception of galactose depressed the enzyme formation (Table 4)

Galactose increased the enz~~e activity about 30

~er centAs with a nwnb~r of enzymes formation of LSD

is also inhibited by the addition of sugarsparticulary

glucoseThe published results of BOYD and LIQISTEIN

(105l)BOYD and LICHSTEIN (1955) EPPS and GALE (1942)

showing that LSD activity is de~ressed in various

cultures grown in a medium containing gluco~eAlthough

carboxyLic acids increased the growth rate of the culshy

turethey inhibited the enzyme formation (~~)le 5)

As a result of this study -Ie found that the be~t

mediurn for the production of LSD by P aeruginos a was

1 ~~ Beef extracb 03 I~2IIPO1 001 IgSObull 7 H 20

and O 5 ~~ nI~Cl

94

Table 3 Effect of amino acids on the fonnation xof L-serine dehydratase

Amino acid L-SDH IUmg Dry wt of cells ~( 05 dry t of cells mgml

None L-serine L-glutaTlic acid Lthreonine L-alanine L-tryptophane L-asparagine L-leucine Glycine

0lG8 OOGO 0043 0082 0102 0041 0164 0107 1160

0648 1200 1220 1140 1100 1250 1120 1300 1320

XThe basal mediQTl contained 1 Beef extract03 K IIP04 0 0 1 HgS047H20and 05 NlI4Cl2

T~~le 4 Effect of carbon canpounds on the fornation of L-serine dehydratasex

Control(no C added) P033 0723 Glucose 0011 1188 Fructose 0014 1228 Haltose 0035 0762 Galactose 0043 0644 Sucrose 0036 0696 Lactose 0034 0716 Dextrine 0036 0776 x The medium contained 1 peptone03 KiRP04and 1

carboxylic acid

Table 5Effeci of carboxylic acids on the formation of L-serine dehydrataseJ~

Carboxylic acid L-SIII ITJmg

dry wt of cells Dry wt of cells

mglml

Control(no added) 004~ 054 Lactate 0007 112 0( -etoglutaratp 0017 094 lUJilrate Ooo~ 001 ri11ltoc OOO~ ll~

ryr_lVutc Ci--r--c

0008 cal~

1O~

072

1Tno _-1~_~lC middot~~~iti comiddot~~tmiddot(J~ 1 ~-c__)~10 I middot 2 -~ ~ e shyl~_ curho~ylic aciL~

95

ACKNOI1LEDGlmiddotlEHTS

Ilinh to thank LHDrooffi for helpful c1iacussions

and for assistance in preparing the manuscript

OZET

paeruginosada L-serin dehidr-atac a]~tivitesi

9alJ~JldJAmonyum ~lorUrUn enzim akt LvL tesini arttJr shydJjJde~isecti1 karbon 1aynaklarJnJn ise Lnh ibo ettii

gOzlendi Manuscript aeceived in April 1980

REFERENCES

lDOYD HL and LICHSTEIN HC (1951) The inhibitory

effect of glucose on certain a~ino acid ceaminases

JBacterio162711-715

2BOYDWLLIaISTEINHC(1955)The influence of nutshy

rition on the serine and threonine dea~in~ses of

microorganisrnsJnacteriol 69545-5middot18

3-EPPSIIlmiddotR and GALEEF(19middot12)The influence of

the 9resence of glucose during groh on the

enzymic activi ties of Es cherichia coli comparLs on

of the effect with that produccc l)y fermentation

acidsBiochem J(London) 36619-623

11 FRIEDErlMN T E and HAUGEN GE (1943) Pyruvic acid

IIThe detennination of ]eto acdds in blood and

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