Aknowledgemen s
This esea ch, is unded by he Fo ma ion and Pe ec ion o he Resea che
Pe sonal P og am o he Basque Coun y Go e nmen , he EXZOME p ojec (CTM
2008 – 01616/MAR) and he CAMVALEX (CTM 10-09515-F) om he Spanish
Science and Educa ion Minis y.
Re e ences
Low y, O. H., N. J. Roseb ough, e al. (1951). "P o ein measu emen wi h he olin phenol eagen ." Jou nal o Biological Chemis y 193: 265 - 275.
Packa d, T. T., E. Be dale , e al. (1996). "Oxygen consump ion in he ma ine bac e ium Pseudomonas nau ica p edic ed om ETS ac i i y and bisubs a e enzyme kine ics." Jou nal o Plank on
Resea ch 18(10): 1819 – 1835.
Packa d and Gomez (2008). "Explo ing a i s -p inciples-based model o zooplank on espi a ion." ICES Jou nal o Ma ine Science 65(3): 371 - 378.
Wagne , T.C., Sco , M.D. (1994). “Single ex ac ion me hod o he spec opho ome ic quan i ica ion o oxidized and educed py idine nucleo ides in e i ocy es”. Analy ical Biochemis y 222: 417-426
Respi a ion
Oxygen elec odes
P o ein
(Low y e al., 1951)
ETS ac i i y
(Packa d e al.,1996)
Py idine
nucleo ides
(Wagne and Sco ,
1994)
P elimina y esul o espi a o y kine ics in zooplank on
samples: MALASPINA 2010
N. Osma a*, T. Packa d a, M. Gómez a
a Ins i u e o Oceanog aphy and Global Change, Uni e si y o Cana y Islands. Cana y Islands, Spain.
* E–mail: nosma@beca ios.ulpgc.es
Fig 4. Analy ical me hods
Fig 5. Linewea e -Bu k ecip ocal
plo o calcula ing he kine ic
pa ame e s in bisubs a e eac ions.
B/O Hespé ides
Fig 3. 100 µm mesh
size WP-2 ne used
o sampling.
Fig 1. C uise ack o he R/V Hespe ides du ing he LEG 3, 4 and 7.
Fig 2. Size ac iona ion and on boa d incuba ion sys em.
Py idine Nucleo ides
Fig 6. P o ein speci ic NADH and NADPH le els in well- ed and s a ed zooplank on. The speci ic concen a ion o bo h NADH and NADPH we e no signi ican ly
di e en in well- ed zooplank on (p>0.001) o he h ee size ac ions. Le els o NADH we e only signi ican ly di e en be ween well- ed and s a ed condi ions in 100-
500 µm size ac ion.
100-500 µm 500-1000 µm > 1000 µm
Table II. Kine ic cons an s o NADH and NADPH in wo size ac ions o zooplank on, The e a e no signi ica i e
di e ences be ween he equilib ium cons an (Km) and he appa en dissocia ion cons an (Kia) o NADH and
NADPH when well ed and s a ed o ganisms a e compa ed. The maximum eloci y (Vmax) o he eac ion
dec eases in s a ed zooplank on. Da a a e gi en as mean ± SD. Numbe s in pa en heses ep esen he numbe
o da a used o calcula e he a e age alues.
The espi a o y oxygen consump ion (R) is a physiological p ocess ha occu s
whe e e he oxygen and he o ganic ma e a e p esen . I is ca alyzed by he
enzyma ic ac i i y o he espi a o y elec on ans e sys em (ETS). This ac i i y, in
u n, is con olled by he a ailabili y o subs a es, i.e., nico inamide adenine
dinucleo ide (NADH) and nico inamide adenine dinucleo ide phospha e (NADPH).
The Enzyme Kine ic Model (EKM) a gues ha , acco ding o he heo y o
Michaelis- Men en, he maximum ac i i y o hese enzymes and he a ailabili y o
hei subs a es con ol he in i o ac i i y o he ETS enzymes a cons an
empe a u es (Packa d and Gomez, 2008). Up o now, no s udy has measu ed he
NADH and NADPH in acellula le els and applied hem o he EKM in he
zooplank on. In he p esen wo k, we ha e measu ed he in acellula le el o
subs a es and calcula ed he kine ics cons an s.
Field samples we e collec ed du ing h ee legs o he Malaspina 2010
oceanog aphic c uise (Fig 1). The physiological measu emen o espi a ion we e
accomplished on boa d in well ed and in 24h s a ed o ganisms. The enzyme
ac i i ies and kine ics and in acellula subs a es le els we e de e mined back a
labo a o y.
INTRODUCTION
I. NADH le els emained nea ly cons an om well- ed o s a a ion condi ions in 500-1000 µm and >1000 µm size ac ions. Howe e , hey
dec eased in 100-500 µm size ac ion. Highes size classes migh ha e a la ge ese e and 24 h o s a a ion migh no challenge hem.
II. Speci ic espi a o y oxygen consump ion dec eases wi h size ac ion, bo h in well- ed and s a a ion condi ions.
III. The e a e no signi ican di e ences in he kine ics cons an s be ween well- ed and s a ed o ganisms.
CONCLUSSIONS
Table I. P o ein speci ic a es o espi a o y oxygen consump ion (R) and po en ial espi a ion (Φ) bo h in
well- ed and in s a ed zooplank on. Da a a e gi en as means ± SD.
ENZYME KINETIC MODEL
Ro =
Vmax: Maximum eloci y o he eac ion
S1, S2: Subs a e concen a ion o NADH and NADPH
K1, K2: Michaelis cons an o NADH and NADPH
Kia: Appa en dissocia ion cons an
𝑉𝑉𝑉𝑉 · [𝑆𝑆 ·𝑆𝑆]
𝐾𝑆 · 𝐾𝐾𝑉 + 𝐾𝑆 · [𝑆𝑆] + 𝐾𝑆 · [𝑆𝑆] + [𝑆𝑆 ·𝑆𝑆]