Vol.
44,
No.
5
APPLIED
AND
ENVIRONMENTAL
MICROBIOLOGY,
No .
1982,
P.
1020-1025
0099-2240/82/111020-06$02.00/0
Copy igh
©
1982,
Ame ican
Socie y
o
Mic obiology
Sus ained
Pho op oduc ion
o
Ammonia
om
Ni a e
by
Anacys is
nidulans
JUAN
L.
RAMOS,
MIGUEL
G.
GUERRERO,*
AND
MANUEL
LOSADA
Depa amen o
de
Bioquimica,
Facul ad
de
Biologia
y
Consejo
Supe io
de
In es igaciones
Cien i icas,
Uni e sidad
de
Se illa,
Apa ado
1095,
Se illa,
Spain
Recei ed
9
Ma ch
1982/Accep ed
19
July
1982
Condi ions
ha
leng hen
he
ime
du ing
which
L-me hionine-DL-sul oximine
(MSX)
p omo es
exc e ion
o
ammonia
p oduced
by
pho osyn he ic
ni a e
educ ion
in
Anacys is
nidulans
ha e
been
sough .
I
MSX
was
added
e e y
24
h,
maximal
a es
o
ammonia
p oduc ion
we e
main ained
o
3
days.
A e
his
ime,
ammonia
p oduc ion
ceased
due
o
a
speci ic
de iciency
o
glu amine
in
he
cells,
which
inally
led
o
cell
lysis.
The
e ec i e
ammonia
p oduc ion
pe iod
could
be
u he
ex ended
ei he
by
adding
a
low
amoun
o
glu amine
a
he
end
o
he
3-
day
pe iod
o
by
allowing
he
cells
o
eco e
o
8
h
in
he
absence
o
MSX
a e
e e y
48-h
pe iod
in
he
p esence
o
inhibi o .
In
his
way,
a
s eady
p oduc ion
o
ammonia
las ing
o
a
leas
10
days
was
achie ed.
The
MSX- ea ed
cyanobac e-
ial
cells
hus
ep esen
a
sys em
ela i ely
s able
wi h
ime
o
he
con e sion
o
ligh
ene gy
in o
chemical
ene gy
h ough
he
pho o educ ion
o
ni a e
o
ammonia.
The
pho o educ ion
o
ni a e
o
ammonia
by
pho osyn he ic
o ganisms
ep esen s
a
ema k-
able
p ocess
o
he
con e sion
o
sola
ene gy
in o
chemical
ene gy
(4,
7,
8).
E icien
pho o-
p oduc ion
o
ammonia
om
ni a e
by
cells
o
he
cyanobac e ium
Anacys is
nidulans
has
been
achie ed
in
ou
labo a o y
a e
speci ically
in-
hibi ing
glu amine
syn he ase,
he
i s
enzyme
in ol ed
in
he
main
ammonia
assimila ion
pa h-
way
in
his
o ganism,
wi h
he
glu ama e
analog
L-me hionine-DL-sul oximine
(MSX)
(3,
11).
To
be
conside ed
aluable
o
p ac ical
pu -
poses,
a
sys em
o
he
con e sion
o
sunligh
ene gy
mus
ope a e
a
conside able
a e
and
e iciency
and
mus
be
ela i ely
s able
in
ime.
The
sys em
cons i u ed
by
MSX- ea ed
Ana-
cys is
cells
has
been
shown
o
p oduce
ammonia
a
high
a es
(11)
wi h
ela i ely
high
e iciency
(12),
bu
un il
now
he
p ocess
has
been
only
ca ied
ou
du ing
a
limi ed
pe iod
o
ime.
This
communica ion
epo s
he
achie emen
o
a
s eady
pho op oduc ion
o
ammonia
om
ni a e
by
MSX- ea ed
A.
nidulans
cell
suspen-
sions,
las ing
o
a
pe iod
o
a
leas
10
days.
MATERIALS
AND
METHODS
G ow h
o
cells.
A.
nidulans
1402-1
was
g own
pho oau o ophically
unde
con inuous
ligh ing
(Syl-
ania
dayligh
luo escen
ubes,
gi ing
an
i adiance
alue
o
25
W/m2
a
he
su ace
o
he
cul u e
essels),
a
40°C
on
a
syn he ic
medium
con aining
ni a e
as
he
ni ogen
sou ce
(5),
in
a
s eam
o
2%
CO2
in
ai
( ol/
ol),
which
was
bubbled
h ough
he
cul u e
a
a
low
a e
o
1
li e /li e
o
cell
suspension
pe
h.
Ammonia
p oduc ion
expe imen s.
Cells
om
2-day
old
cul u es
con aining
20
o
25
,ug
o
chlo ophyll
a
(Chl)
pe
ml
we e
used.
The
cells
we e
ha es ed
by
cen i uga ion,
washed
wi h
cul u e
medium,
and
inal-
ly
suspended
in
he
same
medium
o
a
densi y
o
8
o
10
,Lg
o
Chl
pe
ml.
MSX
was
added
o
he
cell
suspension
o
a
inal
concen a ion
o
10
FM,
and
he
cell
suspensions
we e
hen
incuba ed
unde
s anda d
cul u e
condi ions
wi h
he
indica ed
modi ica ions.
Assays
o
enzyme
ac i i ies.
Ni a e
educ ase
and
glu amine
syn he ase
( ans e ase)
ac i i ies
we e
measu ed
in
oluenized
cells
as
p e iously
desc ibed
(11).
Ni i e
educ ase
and
NADH-alanine
dehyd o-
genase
we e
de e mined
in
cell- ee
ex ac s.
Ni i e
educ ase
ac i i y
was
es ima ed
acco ding
o
Man-
zano
e
al.
(10).
NADH-alanine
dehyd ogenase
was
es ima ed
as
desc ibed
by
Rowell
and
S ewa
(13).
Ex ac ion
o
cellula
me aboli es.
Pe chlo ic
acid
was
added
o
a
sample
o
he
cell
suspension
o
gi e
a
concen a ion
o
0.6
N.
A e
10
min
a
0°C,
he
samples
we e
cen i uged
a
40,000
x
g
o
15
min.
The
supema an
was
neu alized
a
0°C
wi h
KOH.
A e
emo al
o
he
KC104
p ecipi a e
by
cen i uga ion
(10,000
x
g
o
10
min),
he
supema an s
we e
lyophi-
lized.
The
lyophilized
ma e ial
was
suspended
o
a
small
olume
wi h
wa e
and
used
o
he
es ima ion
o
me aboli es
(3).
Analy ical
me hods.
Ni a e,
ni i e,
ammonia,
-y-
glu amyl
hyd oxama e,
glu ama e,
and
glu amine
we e
es ima ed
as
desc ibed
p e iously
(11).
Ca bohyd a es
we e
es ima ed
by
he
phenol-sul u ic
me hod
(6),
o al
ni ogen
by
he
Kjehldal
p ocedu e,
packed
cell
ol-
ume
by
cen i uga ion
o
he
cell
suspensions
in
cali-
b a ed
hema oc i
ubes,
and
cellula
p o ein
by
a
modi ica ion
o
he
Low y
p ocedu e
(2)
a e
p e ea -
ing
he
cells
wi h
10%
(w / ol)
ichlo oace ic
acid.
Chl
was
de e mined
spec opho ome ically
in
me hanolic
1020
on June 23, 2015 by USE/BTCA.GENERAL UNIVERSITARIA Se illah p://aem.asm.o g/Downloaded om
SUSTAINED
AMMONIA
PHOTOPRODUCTION
1021
0
au_o_
-°
o
e
o
I
0
D.
Z
20
40
(
h)1.5w
z
0~~~05
~~~~~~z
FIG
1.
Tiecus
ee nslaig m
ea ed
AnuncsAe
00
i
10
Fd-
E~~~~~~~~A
~~~~~~~~~~~10
0~~~~~~~~~
:
cod osDape
eew dana
h
ie
indc d
an
amoi
n h
eim(),dniyo
C
-A-A
~A-A-A
A-------
_--
0
20
40
TIME
(h)
FIG.
1.
Time
cou se
o
he
e en s
leading
o
am-
monia
p oduc ion
and
o
i s
la e
cessa ion
in
MSX-
ea ed
A.
nidulans
cells.
A
cell
suspension
(150
ml)
wi h
a
densi y
o
9
,ug
o
Chl
pe
ml
and
supplemen ed
wi h
10
pe M
MSX
was
incuba ed
unde
s anda d
cul-
u e
condi ions.
Samples
we e
wi hd awn
a
he
imes
indica ed,
and
ammonia
in
he
medium
(6),
densi y
o
he
cell
suspension
(A),
and
cellula
ac i i y
le els
o
ni a e
educ ase
(0)
and
glu amine
syn he ase
(A)
we e
de e mined.
ex ac s
wi h
he
ex inc ion
coe icien
gi en
by
MacKinney
(9).
The
Chl
and
p o ein
con en s
o
MSX-
ea ed
A.
nidulans
cells
we e
ai ly
cons an ,
wi h
alues
o
5.8
o
6.2
pLg
o
Chl
and
110
o
130
p.g
o
p o ein
pe
dJ
o
packed
cell
olume
(11).
Measu e-
men s
o
he
abso bance
alues
a
628
and
680
nm
o
he
cell
suspensions
o
es ima ion
o
he
phycocyaninl
Chl
a io
we e
pe o med
by
placing
he
sample-
con aining
cu e e
in
he
u bid-samples
compa men
o
a
Pye
Unicam
SP
1750
spec opho ome e .
RESULTS
AND
DISCUSSION
The
main
e en s
aking
place
a e
he
addi-
ion
o
MSX
o
suspensions
o
A.
nidulans
cells
in
ni a e
medium
ha e
p e iously
been
de-
sc ibed
(11)
and
can
be
summa ized
as
ollows.
Glu amine
syn he ase
becomes
apidly
and
com-
ple ely
inac i a ed,
and
ammonia
appea s
in
he
medium
concomi an ly
wi h
his
ac i i y
loss.
Cell
g ow h
is
p e en ed,
and
he
a es
o
ni a e
up ake
and
educ ion
inc ease
abou
wo old.
Ammonia
accumula es
in
he
medium
a
a
con-
s an
a e
o
abou
25
pLmolUmg
o
Chl
pe
h.
The
p ocess
o
ammonia
p oduc ion
con inues
o
abou
30
o
35
h
wi hou
u he
addi ions,
p o ided
ha
ni a e
is
no
limi ing,
and
ni a e
educ ase
ac i i y
emains
a
i s
o iginal
high
le el
du ing
his
pe iod.
As
Fig.
1
shows,
a e
his
ime,
glu amine
syn he ase
ac i i y
eco -
e ed,
and
he
accumula ion
o
ammonia
in
he
medium
ceased,
ollowed
by
i s
u iliza ion
by
he
cells,
which
s a ed
o
g ow.
Once
glu amine
syn he ase
became
ac i e,
he
an agonis ic
e -
ec s
o
ammonia
(3,
5,
11)
came
in o
play;
ni a e
u iliza ion
was
inhibi ed
(da a
no
shown),
and
he
ni a e
educ ase
le el
de-
c eased
apidly
(Fig.
1).
Since,
unde
s anda d
cul u e
condi ions
and
in
he
absence
o
cells,
MSX
does
no
lose
i s
ac i i y
o
a
leas
48
h,
he
cessa ion
o
i s
e ec
on
ammonia
p oduc-
ion
can
be
ela ed
o
he
ans o ma ion
o
he
inhibi o
by
he
algal
cells
a he
han
o
i s
spon aneous
deg ada ion.
I
seems,
he e o e,
ha
he
empo a y
ceas-
ing
o
ammonia
pho op oduc ion
is
due
o
he
eco e y
o
glu amine
syn he ase
ac i i y
a he
han
o
he
induc ion
o
any
o he
ammonia
assimila ion
pa hway.
Thus,
o
leng hen
he
pe-
iod
o
ammonia
p oduc ion,
esh
MSX
a
i s
op imal
concen a ion
wi h
espec
o
he
densi-
y
o
he
cell
suspension
(1
nmol/,lg
o
Chl)
(12)
was
added
o
he
Anacys is
cell
suspension
in
which
he
e ec
o
he
o iginal
MSX
had
been
los .
This
addi ion
esul ed
again
in
inac i a ion
o
he
p eexis ing
glu amine
syn he ase,
allow-
ing
new
maximal
a es
o
ammonia
p oduc ion
(da a
no
shown).
Following
his
app oach,
successi e
addi ions
o
10
,uM
MSX
o
he
cell
suspension
e e y
24
h
allowed
maximal
a es
o
ammonia
p oduc ion
o
be
sus ained
o
3
days.
Du ing
his
pe iod,
cellula
glu amine
syn he ase
emained
ully
inac i e,
he
ni a e
educ ase
ac i i y
le el
being
high,
and
abou
90%o
o
he
ni a e
aken
up
appea ing
in
he
medium
as
ammonia
(Fig.
2).
A
his
s age,
he
addi ion
o
MSX
no
longe
esul ed
in
main enance
o
he
p ocess
o
ammonia
p oduc ion,
and
cell
lysis
o
he
suspension
was
obse ed
sho ly
a e wa ds.
To
de e mine
he
cellula
si ua ion
leading
o
he
cessa ion
o
ammonia
p oduc ion
by
Anacys-
is,
he
le els
o
a ious
cell
componen s,
enzy-
ma ic
ac i i ies
ela ed
o
ni a e
me abolism,
and
amino
acids
in ol ed
in
he
glu amine
syn-
he ase-glu ama e
syn hase
pa hway
we e
s ud-
ied
in
Anacys is
cells
which
had
been
ea ed
wi h
MSX
o
68
h
and
we e
compa ed
wi h
hose
in
no mal
cells.
The
analysis
o
Anacys is
cells
subjec ed
o
MSX
ea men
o
68
h
yield-
ed
da a
simila
o
hose
p e iously
ound
in
expe imen s
o
sho e
du a ion
(11).
The
cellu-
la
le els
o
p o ein,
Chl,
phycocyanin,
and
o al
ni ogen
emained
cons an
and
equal
o
hose
o
un ea ed
cells,
whe eas
he
ca bohyd a e
le el
inc eased
linea ly.
The
inc ease
in
he
ca bohy-
d a e
le el,
om
6.2
,ug/,ul
o
cells
a
ime
ze o
o
39
pug/Ll
o
cells
a
68
h,
was
in e p e ed
as
due
o
con inued
pho osyn he ic
CO2
ixa ion
while
VOL.
44,
1982
on June 23, 2015 by USE/BTCA.GENERAL UNIVERSITARIA Se illah p://aem.asm.o g/Downloaded om
1022
RAMOS,
GUERRERO,
AND
LOSADA
0
w
w
a.
a-
z
_
CZ
E
o
W
w1
E
C.)
=.
0
0
a-
z
0
4
C1
2-
N
w
Ln
w
4
z
20
C-
8
E
"I
w
0
z
10
w
z
Ln
zS
I.
D
O
(D
24
48
72
TIME
(h)
FIG.
2.
Con inuous
ammonia
pho op oduc ion
by
MSX- ea ed
A.
nidulans
cells
o
3
days.
Condi ions
we e
hose
desc ibed
in
he
legend
o
Fig.
1,
excep
ha
MSX
(1.5
,umol)
was
added
e e y
24
h.
A
48
h,
KNO3
(1.5
mmol)
was
added.
Samples
we e
wi hd awn
a
he
imes
indica ed,
and
ammonia
(0)
and
ni a e
(0)
in
he
medium
and
cellula
ac i i y
le els
o
ni a e
educ ase
(A)
and
glu amine
syn he ase
(A)
we e
de e mined.
MSX
p e en ed
he
inco po a ion
o
ammonia
in o
ca bon
skele ons
and
subsequen
cell
g ow h
(11).
I
is
in e es ing
ha
he
phycocyanin
le els
ypical
o
no mal
cells
we e
main ained
in
MSX-
ea ed
Anacys is
cells
when
he
medium
con-
ained
ni a e
bu
no
when
i
was
ee
o
ni o-
1
6E
7
16
U
w
A
gen.
As
a
consequence,
MSX- ea ed
Anacys is
cells
ac i ely
p oducing
ammonia
om
ni a e
did
no
su e
enhanced
phycocyanin
deg ada-
ion,
as
is
he
case
o
cells
subjec ed
o
ni ogen
s a a ion
(1).
This
con as s
wi h
he
si ua ion
obse ed
o
MSX- ea ed
ilamen ous
cyano-
bac e ia
unde
dini ogen- ixing
condi ions,
as
4
Sn
E
^
~~~~~2
A
~~~w
z
A
I-
D
0
0o
30
60
TIME
(
h
)
FIG.
3.
Cellula
le els
o
glu amine
and
glu ama e
in
MSX- ea ed
and
un ea ed
A.
nidulans
cells.
Cells
g own
on
ni a e
medium
we e
ans e ed
o
media
con aining
KNO3
(20
mM)
(open
symbols)
o
KNO3
(20
mM)
and
MSX
(10
FM)
(closed
symbols).
Samples
we e
wi hd awn
a
he
imes
indica ed,
and
he
cellula
con en
in
glu amine
(ci cles)
and
glu ama e
( iangles)
was
es ima ed.
O he
condi ions
we e
hose
desc ibed
in
he
legend
o
Fig.
1.
APPL.
ENVIRON.
MICROBIOL.
on June 23, 2015 by USE/BTCA.GENERAL UNIVERSITARIA Se illah p://aem.asm.o g/Downloaded om
SUSTAINED
AMMONIA
PHOTOPRODUCTION
30
1-
23
1-
16
31
O
c
0~~~~~
0~~~~~~~
0
0
_ o
--
o-o
W8e"
I,o
o/~~~~
IA
,.o
I/0
72
96
10
48
TIME
(h)
FIG.
4.
E ec
o
glu amine
addi ion
on
he
p olonga ion
o
he
ammonia
p oduc ion
pe iod
by
MSX- ea ed
A.
nidulans
cells.
Condi ions
we e
hose
desc ibed
in
he
legend
o
Fig.
2,
excep
ha
glu amine
(0.5
mM)
was
added
a
70
h
(a ow)
o
one-hal
o
he
suspension
(0);
he
o he
hal
was
used
as
a
con ol
wi hou
any
addi ion
(0).
ound
o
Anabaena
cylind ica
(14)
and
Ana-
baena
sp.
s ain
ATCC
33047
(unpublished
da a).
Al hough
MSX- ea ed
Anacys is
cells
lacked
glu amine
syn he ase,
hey
exhibi ed
low
bu
ep oducible
le els
o
NADH-alanine
dehyd o-
genase
(a ound
7
mU/mg
o
p o ein),
le els
o
he
same
o de
o
magni ude
as
hose
ound
in
no mal
cells.
This
inding
sugges s
ha
ammonia
assimila ion
pa hway(s)
ha
may
be
seconda y
unde
no mal
condi ions,
can
play
an
impo an
ole
in
he
main enance
o
he
ni ogen
balance
in
MSX- ea ed
cells.
The
enzymes
esponsible
o
ni a e
educ ion,
namely,
ni a e
educ ase
and
ni i e
educ ase,
in
MSX- ea ed
Anacys is
cells
we e
always
p esen
a
abou
he
same
le els
as
in
he
un ea ed
cells.
As
a
consequence
o
he
inac i a ion
by
MSX
o
glu amine
syn he ase,
he
glu amine
le el
in
ammonia-p oducing
cells
dec eased
apidly
wi h
ime
and
became
p ac ically
negligible
30
h
a e
he
addi ion
o
he
inhibi o .
This
beha io
was
in
con as
wi h
ha
o
un ea ed
cells,
which,
unde
he
same
condi ions,
main ained
hei
no mal
glu amine
le els.
The
le el
o
glu ama e
in
MSX- ea ed
Anacys is
cells
also
dec eased
wi h
ime,
bu
much
slowe ;
i s
alue
a e
68
h
was
35%
ha
o
no mal
cells
(Fig.
3).
This
amino
acid
analysis
indica es
ha
Anacys is
cells
sub-
jec ed
o
ea men
wi h
MSX
become
speci i-
cally
de icien
in
glu amine
and
p obably
also
in
glu amine
de i a i es.
Such
a
me abolic
si ua-
ion
migh
e en ually
lead
o
he
cessa ion
o
ammonia
p oduc ion.
To
o e come
he
MSX-p omo ed
glu amine
de iciency
and
hus
p olong
he
ammonia
p o-
duc ion
pe iod,
wo
di e en
a emp s
we e
made.
One
o
hem
consis ed
o
he
addi ion
o
he
cell
suspension
o
glu amine
a
low
concen-
a ions
(0.2
o
0.5
mM)
simul aneously
wi h
he
no mal
eaddi ion
o
MSX
a
he
end
o
he
3-
day
pe iod.
The
da a
in
Fig.
4
show
ha
his
glu amine
ea men
esul ed
in
sus ained
ammo-
nia
p oduc ion
a
cons an
a e
o
a
leas
24
h,
a
phenomenon
ha
did
no
occu
in
a
con ol
wi hou
glu amine
addi ion,
in
which
he
a e
o
ammonia
p oduc ion
slowed
wi h
ime
un il
i
became
negligible.
In e es ingly
enough,
he
glu-
amine
ea men
did
no
esul
in
cell
g ow h
(measu ed
as
Chl)
o
in
any
inc ease
in
glu a-
mine
syn he ase
ac i i y,
bu
he
le els
o
ni a e
and
ni i e
educ ases
doubled.
No
inc emen
in
he
ammonia
p oduc ion
a e
was
obse ed,
howe e ,
sugges ing
ha
he
no mal
ni a e
and
ni i e
educ ase
le els
a e
high
enough
o
main-
ain
he
p ocess
a
i s
maximal
a e.
Glu ama e
could
no
eplace
glu amine
in
sus aining
ammo-
nia
p oduc ion,
a
esul
which
con i ms
he
assump ion
ha
he
cessa ion
o
he
p ocess
is
speci ically
ela ed
o
glu amine
de iciency.
I
E
I
:3
w
I
z
z
0
I
I
1023
VOL.
44,
1982
on June 23, 2015 by USE/BTCA.GENERAL UNIVERSITARIA Se illah p://aem.asm.o g/Downloaded om
1024
RAMOS,
GUERRERO,
0
E
E
0
w
0
0
a-
4
z
0
-J
I-
0
6
3
0
100
200
TIME
(
h
)
FIG.
5.
E ec
o
he
ecu en
addi ion
and
emo al
o
MSX
on
he
pho op oduc ion
o
ammonia
by
A.
nidulans
cells.
A
cell
suspension
wi h
a
densi y
o
9
p.g
o
Chl
pe
ml
was
main ained
o
48
h
in
ni a e
cul u e
medium
supplemen ed
wi h
10
puM
MSX.
A e
his
ime,
cells
we e
ha es ed
and
washed
h ee
imes
wi h
esh
ni a e
medium
lacking
MSX.
A e
8
h
in
he
absence
o
he
inhibi o ,
and
once
he
cell
loading
had
been
co ec ed
o
9.0
+
0.2
p.g
o
Chl
pe
ml,
MSX
(10
puM
inal
concen a ion)
was
eadded.
Successi e
pe iods
o
ammonia
p oduc ion
(a ound
48
h)
and
cell
eco e y
(a ound
8
h)
(a ows)
we e
al e na ed
as
indica ed.
The
las
eaddi ion
o
MSX
was
o
each
a
inal
concen a ion
o
100
puM.
The
second
app oach
o
ex end
he
ammonia
p oduc ion
pe iod
consis ed
o
allowing
he
cells
o
eco e
om
glu amine
de iciency
by
hem-
sel es.
This
goal
was
achie ed
by
al e na ing
exposu e
o
MSX
wi h
pe iods
in
which
he
glu amine
syn he ase
inac i a o
was
emo ed.
Figu e
5
shows
ha
his
sequen ial
ea men
allowed
con inuous
p oduc ion
o
ammonia
a
high
a es
o
a
leas
10
days.
The
long
pe iod
in
he
p esence
o
MSX
(a ound
48
h)
esul ed
in
ne
ammonia
p oduc ion,
whe eas
he
sho e
pe iod
in
i s
absence
(a ound
8
h)
did
no .
Du ing
he
eco e y
pe iods,
glu amine
syn he ase
ac-
i i y
inc eased
om
0
o
150
o
300
mU/mg
o
p o ein
(10
o
2b%
o
he
le el
in
no mal
cells),
which
allowed
glu amine
syn hesis
and
a
sligh
g ow h
( om
9.0
o
abou
10.6
,ug
o
Chl
pe
ml
in
8
h).
Su p isingly,
a e
day
8
o
ammonia
p oduc ion,
esis ance
o
MSX
appea ed.
This
phenomenon
was
p obably
due
o
an
adap a ion
o
he
cells
o
MSX,
making
i
necessa y
o
inc ease
he
MSX
concen a ion
up
o
100
,uM
o
again
achie e
comple e
inac i a ion
o
cellula
glu amine
syn he ase.
LITERATURE
CITED
1.
Alien,
M.
M.,
and
A.
I.
Smi h.
1%9.
Ni ogen
chlo osis
in
blue-g een
algae.
A ch.
Mik obiol.
69:114-120.
2.
Bailey,
J.
L.
1%7.
Techniques
in
p o ein
chemis y,
2nd
ed.,
p.
340.
Else ie ,
Ams e dam.
3.
Flo es,
E.,
M.
G.
Gue e o,
and
M.
Losada.
1980.
Sho -
e m
ammonium
inhibi ion
o
ni a e
u iliza ion
by
Ana-
cys is
nidulans
and
o he
cyanobac e ia.
A ch.
Mic obiol.
128:137-144.
4.
Gue e o,
M.
G.,
J.
M.
Vega,
and
M.
Losada.
1981.
The
assimila o y
ni a e- educing
sys em
and
i s
egula ion.
Annu.
Re .
Plan
Physiol.
32:169-204.
5.
He e o,
A.,
E.
Flo es,
and
M.
G.
Gue e o.
1981.
Regula-
ion
o
ni a e
educ ase
le els
in
he
cyanobac e ia
Ana-
cys is
nidulans,
Anabaena
sp.
s ain
7119,
and
Nos oc
sp.
s ain
6719.
J.
Bac e iol.
145:175-180.
6.
Koche ,
G.
1978.
Ca bohyd a e
de e mina ion
by
he
phenol-sul u ic
me hod,
p.
95-97.
In
J.
A.
Hellebus
and
J.
S.
C aigie
(ed.),
Handbook
o
phycological
me hods.
Physiological
and
biochemical
me hods.
Camb idge
Uni-
e si y
P ess,
Camb idge,
England.
7.
Losada,
M.
1979.
Pho op oduc ion
o
ammonia
and
hyd o-
gen
pe oxide.
Bioelec ochem.
Bioene g.
6:205-225.
8.
Losada,
M.,
and
M.
G.
Gue e o.
1979.
The
pho osyn he -
ic
educ ion
o
ni a e
and
i s
egula ion,
p.
366-403.
In
J.
Ba be
(ed.),
Pho osyn hesis
in
ela ion
o
model
sys ems.
Else ie ,
Ams e dam.
9.
MacKinney,
G.
1941.
Abso p ion
o
ligh
by
chlo ophyll
solu ions.
J.
Biol.
Chem.
140:315-322.
0
0
/
0-0
0
0/
0-0
/
A0
0
I
--I
1
-I
AND
LOSADA
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ENVIRON.
MICROBIOL.
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SUSTAINED
AMMONIA
PHOTOPRODUCTION
1025
10.
Manzano,
C.,
P.
Candau,
C.
G6mez-Mo eno,
A.
M.
Re-
l inpio,
and
M.
Losada.
1976.
Fe edoxin-dependen
pho-
osyn he ic
educ ion
o
ni a e
and
ni i e
by
pa icles
o
Anacys is
nidulans.
Mol.
Cell.
Biochem.
10:161-169.
11.
Ramos,
J.
L.,
M.
G.
Gue e o,
and
M.
Losada.
1982.
Pho op oduc ion
o
ammonia
om
ni a e
by
Anacys is
nidulans
cells.
Biochim.
Biophys.
Ac a
679:323-330.
12.
Ramos,
J.
L.,
M.
G.
Gue e o,
and
M.
Losada.
1982.
Op imiza ion
o
condi ions
o
pho op oduc ion
o
ammo-
nia
om
ni a e
by
Anacys is
nidulans.
AppI.
En i on.
Mic obiol.
44:1013-1019.
13.
RowelU,
P.,
and
W.
D.
P.
S ewa .
1976.
Alanine
dehyd o-
genase
o
he
N2- ixing
blue-g een
alga,
Anabaena
cylin-
d ica.
A ch.
Mic obiol.
107:115-124.
14.
Wood,
N.
B.,
and
R.
Haselko n.
1980.
Con ol
o
phyco-
bilip o ein
p o eolysis
and
he e ocys
di e en ia ion
in
Anabaena.
J.
Bac e iol.
141:1375-1385.
VOL.
44,
1982
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