RESPIRATION:

Basic mechanisms, factors affecting, measurement

PROGRAM STUDI S2 TEKNOLOGI PASCAPANEN
DEPARTEMEN TEKNIK MESIN DAN BIOSISTEM
INSTITUT PERTANIAN BOGOR
2013
1
Oleh: Dr.Ir. Rokhani Hasbullah, MSi.
Materi Kuliah
TEKNIK PASCAPANEN TANAMAN HORTIKULTURA
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Basic Mechanisms of Respiration
The term of respiration: ... the physical and chemical
processes by which an organism supplies its cells and
tissues with the oxygen needed for metabolism and relieves
them of the carbon dioxide produced in energy-producing
reactions... (Webster).
Biochemists ... Any of various energy-yielding oxidative
reactions in living matter...
Respiration is a vital process to humans and other animals.
It is equally important to plant cells and tissues.
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Respiration The electrical analogy
The metabolic process in living cells is the high energy
stored in the third phosphate bond of adenosine triphosphate
(ATP).
ATP in the cell ===> rechargeable batteries
Batteries contain energy can be liberated to flashlights,
radio, tape recorder, etc.
Unlike batteries, without which our lives would continue
(although much changed), a continued supply of ATP is
absolutely vital to the fundamental processes of life.
Maintaining the supply of ATP is the primary purpose of
respiration.
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Electricity generation
A model for cell respiration
Cell respiration
The TCA cycle furnace (Krebs cycle) must be supplied
with a specific molecular fuel, acetyl CoA.
by the process of glycolysis (sugar splitting)
5
Oksigen akan digunakan untuk mensintesis energi
ATP dalam respirasi aerob.
Pada sel, terjadi dalam mitokondria
NADH = the spark
fuel = glucose
exhaust =
carbon dioxide
air = oxygen
OUTPUT
engine heat
= body heat
INPUT
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A key enzyme in this process is phospho-fructokinase
(PFK), a multicomponent enzyme which converts
fructose-6-phosphate into fructose-1,6-diphosphate,
thereby preparing the hexose for cleavage into 2 triose
phosphate compounds, glyceraldehyde-3-phosphate and
dihydroxyacetone phosphate.
The powerhouse of the cell mitochondrion
Contains nucleic acids
and ribosomes for
synthesis protein and
enzymes.
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Acetyl-CoA
Oxaloacetate Citrate
Malate
Aconitate
Fumarate Isocitrate
Succinate
-Ketoglutarate
Succinyl-CoA
H2O
CO2
TCA Cycle
CoA
CO2
CO2
O2
ADP+P
ATP
Pyruvate
Starch
Glucose-1-P
Glucose-6-P
Fructose-6-P
Fructose-1,6-diP
Phosphoglyceraldehyde
Phosphoenolpyruvate
3-Phosphoglycerate
1,3-Diphosphoglycerate
2-Phosphoglycerate
Lactate
Acetaldehyde
Ethanol
CO2
Fermentation
The Krebs (TCA) cycle
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The basic chemistry of the process, in which the oxidation of
glucose is coupled to regeneration of ATP from ADP
(adenosine monophosphate) and Pi (inorganic phosphate)
with the release of H
2
O and O
2
is:
C
6
H
12
O
6
+ 6O
2
+ 38 ADP + 38Pi => 6CO
2
+ 44 H
2
O + 38 ATP
Respiration is a process whereby the free energy locked
up in glucose is used to generate a membrane potential by
oxidising the glucose molecule to CO
2
and H
2
O.
This membrane potential is used directly to replenish the
cells supply of ATP.
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In the Generator:
The energy produced by fuel combustion is absorbed in
a heat exchanger where water is converted to high
pressure steam.
Steam becomes the energy resource.
In the Respiration:
Oxidation implies the loss of electrons from the fuel
molecule and their donation to an electron acceptor,
which is thereby reduced.
Low energy Nicotinamide adenine dinucleotide (NAD)
is reduced to the high energy form, NADH, thus:
NAD
+
+ H
+
+ 2e (from the fuel) NADH
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The electron transport system
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Concentration of ATP remains constant at all times. The net
effect of respiration can be considered to be the oxidation of
glucose to CO
2
and H
2
O with the liberation of energy:
C
6
H
12
O
6
+ 6 O
2
+ 6 H
2
O => 6 CO
2
+ 12 H
2
O + 673 Kcal
Fase Respirasi
Polisakarida gula sederhana
Gula sederhana asam piruvat
Asam piruvat karbon dioksida + as organik lainnya
12
Glucose
+
Cellular
Respiration
Glucose
+
+
+
Respirasi
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Perubahan yang terjadi pada produk akibat respirasi
setelah dipanen
Perombakan klorofil dan pembentukan karotenoid (warna hijau
menjadi kuning/jingga)
Pelepasan etilen (menjadi agen pemicu proses pematangan
shg buah akan menjadi matang)
Perombakan karbohydrat menjadi gula (rasa menjadi manis)
Pembentukan zat-zat volatil (menimbulkan aroma buah
matang)
Penguapan air dan pelepasan panas
Bila tidak segera dikonsumsi perubahan akan terus
terjadi dan menjadi proses pembusukan
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Faktor-faktor yang mempengaruhi respirasi
1. Internal factors
a. Genotype (Type of commodity)
b. Type of plant part
c. Stage of development at harvest
d. Respiratory substrat
e. Preharvest factor
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Laju respirasi produk hortikultura pada suhu 5 C
Respirasi
(mg CO2/kg-jam)
Rendah 5 - 10 apel, jeruk, anggur, melon, pepaya, nanas
bw putih, bw merah, kentang, ubi jalar
Sedang 10 -20 pisang, mangga, chery, peach, pear, kubis
wortel, ketimun, batang selada, tomat
Tinggi 20 - 40 alpukat, bunga kol, daun selada
Sangat tinggi 40 - 60 brokoli, okra bunga potong
Paling tinggi > 60 asparagus, jamur, bayam, jagung manis,
jagung muda
Kelompok Komoditas
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Jenis komoditi
16
umbi
batang
Buah/bunga
daun
tunas
Respirasi paling tinggi
Respirasi paling rendah
Asal dari produk menentukan sifatnya
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Perubahan fisiko-kimia
(1) Fase pembelahan sel (cell division) dan
pembesaran sel (cell enlargement)
(2) Fase pendewasaan (maturation)
(3) Fase pematangan (ripening)
(4) Fase pelayuan (senescence) dan diikuti fase
pembusukan (deterioration).
Stage of development at harvest
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Respirasi
Non-klimakterik
Respirasi
Klimakterik
Pendewasaan Pematangan Pelayuan
L
a
j
u

r
e
s
p
i
r
a
s
i
Perubahan selama proses pematangan:
Warna
Rasa
Tekstur
Terbentuknya vitamin
Timbulnya aroma khas
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Buah digolongkan menjadi dua kelompok
Buah Klimakterik: pematangan terjadi setelah laju respirasi
mencapai puncaknya
Dipanen tua, lalu proses pematangan akan terjadi dengan
sendirinya
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Buah Non-klimakterik: laju respirasi terus menurun dan tidak
mempunyai puncak
Dipanen setelah terjadi proses pematangan pada pohon
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Buah Klimakterik Buah Non Klimakterik
Apel
Markisa
Alpukat
Pepaya
Pisang
Nangka
Sirsak
Melon
Semangka
Jambu biji
Tomat
Jeruk
Nanas
Anggur
Stroberi
Salak
Ketimun
Cabe
Bawang
Paprika
Kakao
Cherry
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Respiratory Substrat
Substrat gula/karbohidrat:
C
6
H
12
O
6
+ 6 O
2
6 CO
2
+ 6 H
2
O + 675 kal
Substrat gula RQ = 6/6 = 1
Substrat asamorganik RQ < 1
Berapa nilai RQ apabila substratnya Lemak?
Protein?
2
2
O Konsumsi
CO Produksi
RQ Quotient y Respirator
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Preharvest factor
Lokasi tumbuh
Iklim
Musim
Teknik budidaya pemupukan
The respiration of apples grown under conditions
of low Calcium nutrition is considerably higher
than that of apples with adequate supply of that
element.
23 by RKH
2. Environmental factors
a. Temperature
b. Atmospheric composition
i. Oxygen concentration
ii. Carbon dioxide concentration
iii. Ethylene
c. Physical stress
24 by RKH
Increased temperatures cause an exponential rise in the
rate of respiration which is expressed by the Vant Hoff
rule:
The velocity of a biological reaction increases 2 to 3
fold for every 10
o
C rise in temperature.
The temperature coefficient for 10
o
C interval is called the
Q10 can be calculated by measuring the rate of the
reaction (eg. respiration) at two temperatures:
0 10
o
C Q
10
= 2.5 - 3
10 20
o
C Q
10
= 2 2.5
20 30
o
C Q
10
= 2
Berapa kali laju respirasi buah-buahan/sayuran dari
suhu 0
o
C ke suhu 30
o
C ?
Pengaruh Suhu (Temperature effect)
1 2
T T
10
1
2
10
R
R
Q

by RKH 25
Temperature Assumed Q10 Relative velocity
of deterioration
Relative shelf
life
0 1.0 100
10 3.0 3.0 33
20 2.5 7.5 13
30 2.0 15.0 7
40 1.5 22.0 4
Effect of temperature on rate of deterioration.
by RKH 26
Pengaruh Komposisi Gas (Atmospheric composition)
In tissues where metabolism is high, reduced oxygen concentration
increase the respiration rate, as measured by CO2 production.
The low ATP in the cell stimulates glycolysis (PFK is turned on) to
generate a modest amount of ATP and pyruvate NADH formed
during the oxidation of triose phosphate must be reoxidized. This is
achieved by the reductive decarboxylation of pyruvate to ethanol.
C
6
H
12
O
6
+ 2 ADP + 2 Pi => 2C
2
H
5
OH + 2 CO
2
+ 2 ATP
To maintain the supply of ATP at the aerobic rate, 19 times as
many glucose molecules would be needed, and respiration would
increase 19 fold.
There would be substantial accumulation of ethanol, which is
very toxic to plants.
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Pengukuran respirasi
jumlah substrat (gula) yang hilang
jumlah gas O
2
yang digunakan
jumlah gas CO
2
yang dikeluarkan
jumlah panas yang dihasilkan
jumlah energi (ATP) yang dihasilkan
TEKNIK PENGUKURAN RESPIRASI
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Because the rate of respiration is so tightly coupled to
the rate of cell metabolism, measurement of respiration
can afford an easy non-destructive means of monitoring
the metabolic and physiological state of the tissues.
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Menghitung substrat yang hilang
When hexose sugar is the substrate, 180 grams of sugar
are lost for each 264 grams CO
2
produced by the
commodity.
D
loss
= rate of dry weight loss (g/kg.jam)
%D
loss
= persen dry weight loss per jam (%)
R = respiration rate (mg CO
2
/kg.jam)
264
180
1000
R
D
loss

10,000
0.68 x R
%D
loss

Contoh:
Bawang pada suhu 30 oC respirasinya 35 mg CO2/kg.jam.
Berapa kehilangan berat kering setelah penyimpanan selama
satu bulan?
% 1.73 30 x 24 x 0.0024 bulan per losses Weight
% 0.0024
000 , 10
0.68 x 35
10000
0.68 x R
%D
loss


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Cepat persiapannya
Sederhana
Relatif cepat
Tidak dapat diterapkan dalam CAS/MAP
Tidak dapat digunakan untuk periode
penyimpanan lama
Ketelitian data riskan terhadap
kebocoran


Channel tube
Gas sampling port
CO2
O2
Gas feeding
Produce
Glass jar 4.3 L
W
V
t
x
R

by RKH
Closed System:
R : respirasi (ml/kg.jam)
X : konsentrasi O
2
atau CO
2
(desimal)
t : waktu (jam)
V : volume bebas (ml)
W: berat sampel (kg)
Mengukur Respirasi
30
Open System:
Rumit persiapannya
Memerlukan unit pencampur gas
Boros dalam pemakaian gas
Untuk periode penyimpanan lama
Sangat diperlukan dalam perancangan CAS/MAP
W
Qy Gx
R
1 1
1

W
Gx Qy
R
2 2
2

3
3
x
Qy
G
Kesetimbangn O
2
Kesetimbangan CO
2
Kesetimbangan N
2
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CO2
O2
Produk
Glass jar 4.3 L
Gas inlet
O2 : Y1
CO2 : Y2
N2 : Y3
Gas outlet
O2 : X1
CO2 : X2
N2 : X3
Flow in: Q Flow out: G
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UNIT PENCAMPUR GAS
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Komponen pengatur aliran gas
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Respiration jar
GC Integrator
Gas
sampling port
Plastic pipe
Mass flow meter
Air compressor
o Air humidifier
Incubator Toggle valve
Needle valve
Schematic diagram of respiration rate measurement system.
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Analisis Komposisi Gas
Titrasi
Metode Orsat
Oxygen analyzer
Infrared analyzer
Kromatografi gas
Dr. Rokhani - TMB IPB
Dr. Rokhani - TMB IPB 36
KROMATOGRAFI GAS
Kromatografi cara pemisahan campuran yang didasarkan atas
perbedaan distribusi dari komponen suatu campuran. Pemisahan
tersebut didasarkan pada dua fasa larutan, yaitu fase diam
(stationary) dan fase bergerak (mobile).
Keguanaan kromatografi gas (GC):
Pengujian kemurnian zat tertentu
Memisahkan komponen dari campuran
Mengidentifikasi suatu senyawa
Kromatografi gas terdiri gas pembawa (carrier gas), ruang
suntik sampel (injection port), kolom, oven, detektor,
pencatat (recorder), dan panel kontrol.
Dr. Rokhani - TMB IPB 37
Flame Ionization Detector (FID)
Thermal Conductivity Detector (TCD)
Electron Capture Detector (ECD)
Flame Photometric Detector (FPD)
Thermionic Spesific Detector N, P spesific (TSD)
Photo Ionization Detector (PID)
Detektor kromatografi
38
Pengukuran Respirasi
Detektor: TCD
Carrier gas: Helium
Kolom: CTR I
WG-100
Dr. Rokhani - TMB IPB
Pengukuran Etilen
Detektor: FID
Carrier gas: Helium
Gas pembakar H2, Udara
Kolom: HayeSep T
Kolom tempat terjadinya
proses pemisahan karena di
dalamnya terdapat fase diam.
Jenis kolom:
- packed column
- capillary column
Kolom kromatografi
Dr. Rokhani - TMB IPB 39
Dr. Rokhani - TMB IPB 40
Dr. Rokhani - TMB IPB 41
Kromatografi Gas (GC)
Days Hours CO2 (%) O2 (%)
1 8.00 2.3 19.7
10.00 5.3 16.6
2 8.00 2.2 19.6
10.00 4.2 17.3
3 8.00 2.0 19.8
10.00 3.5 18.1
4 8.00 2.1 19.9
10.00 3.3 18.5
5 8.00 2.3 20.0
10.00 4.6 17.4
6 8.00 2.3 20.1
10.00 3.6 18.5
7 8.00 2.3 19.7
10.00 3.1 18.6
8 8.00 2.3 19.7
10.00 2.9 18.8
Sampel : Mangga
Berat : 1.28. kg
Densitas : 1024 kg/m3
Volume stoples : 4300 ml
1. Hitung laju respirasi buah:
Laju produksi CO
2
Laju konsumsi O
2
2. Tentukan nilai RQ
3. Gambarkan grafik laju
respirasi selama
penyimpanan dan jelaskan
pola respirasinya.
TUGAS
42 by RKH
by RKH 43