Download - 2 Medium Dan Kinetik Farmasi
-
8/10/2019 2 Medium Dan Kinetik Farmasi
1/32
1
Fermentation TechnologyOptimasi medium dan kinetics
MK : Biotechnology
Fakultas Farmasi
Alwani Hamad, ST, MScProgram Studi Teknik Kimia
Fakultas Teknik
Universitas Muhammadiyah Purwokerto
-
8/10/2019 2 Medium Dan Kinetik Farmasi
2/32
2
Course content
Persyaratan proses fermentasi
Kondisi dan variable fermentasi
Fermenter/ bioreaktor
Media dan optimasi media untuk fermentasi
Kinetika pertumbuhan bakteri untuk menghitung
hasil produk fermentasi
Downstream process hasil fermentasi
Evaluasi : Ujian (closed book)Buku rujukan:G Rao.2007. Introduction to Biochemical Engineering. Tata Mc Graw- Hill
Publishing Company Limited
Stanbury, Whitaker and Hall, 2003. Principles of Fermentation Technology
Butterworth
-
8/10/2019 2 Medium Dan Kinetik Farmasi
3/32
3
Quantification of Microbial Rates using
optimum medium
-
8/10/2019 2 Medium Dan Kinetik Farmasi
4/32
4
Microbial rates of consumption or production
C, N, P, S source
H2O
H+
O2
heat
product
CO2
biomass
http://rds.yahoo.com/S=96062857/K=smiley/v=2/SID=w/l=II/R=42/*-http://images.search.yahoo.com/search/images/view?back=http%3a//images.search.yahoo.com/search/images%3fp=smiley%26ei=UTF-8%26cop=mss%26tab=3%26b=41&h=1800&w=1800&imgcurl=www.inboundlogistics.com/cgi-script/csPublisher/library/Smiley%2520Face%2520(flat).jpg&imgurl=www.inboundlogistics.com/cgi-script/csPublisher/library/Smiley%2520Face%2520(flat).jpg&name=%3cb%3eSmiley%3c/b%3e+Face+(flat).jpg&p=smiley&rurl=http%3a//www.inboundlogistics.com/articles/features/0102_feature07.shtml&rcurl=http%3a//www.inboundlogistics.com/articles/features/0102_feature07.shtml&type=&no=42&tt=60,000 -
8/10/2019 2 Medium Dan Kinetik Farmasi
5/32
Medium yang dibutuhkan oleh
mikroorganisme
Carbon :molasses, grain, starch, maize, potatos dan cassava,
sucrose, glucose, dan lactoce
Nitrogen : protein dan asam amino (corn steep liquor, soya
beans, fish meal, yeast extract, dll)
Energy source : chemo-organotrophs( carbon source),heterotrophilic (senyawa organik)
Minerals : K,S, Ca, Fe,Zn, dll
Other nutrient like vitamin, calcium pantothenate untuk vinegar
fermentasi
Oxygen/air for aerobic process:
Water (submerged fermentation)
5
-
8/10/2019 2 Medium Dan Kinetik Farmasi
6/32
Persyaratan pemilihan medium
untuk fermentasi skala besar
Murah, udah didapat dan harga dan kualitas
konsisten
Mempunyai produktifitas yang tinggi : dapat
memproduksi jumlah produk maksimum persubstrat yang dikonsumsi
Pertumbuhan pembentukan produk harus
besar
Harus dapat meminimalkan produk samping
6
-
8/10/2019 2 Medium Dan Kinetik Farmasi
7/32
Medium formulation
Dalam fermentasi medium digunakan untuk :
Optimal growth of the cells
Product formation
Medium formulation dalam skala scale up
Economic viability
membutuhkan teknik yang tepat agar fermentasi dapat
berjalan dengan baik
Cells maintances untuk biosynthesis membentuk produk
7
-
8/10/2019 2 Medium Dan Kinetik Farmasi
8/32
Minimum medium dalam
pembentukan alpha amilase
Soluble starch 20 g/l
Glucose 5 g/l
Peptone 5 g/l
Yeast extract 2,5 g/l
K2HPO4 2 g/l
MgSO4. 7H2O 1 g/l
pH 7
Distilled water 100 ml
8
-
8/10/2019 2 Medium Dan Kinetik Farmasi
9/32
Aplikasi penggunaan medium
dalam industri
Substrate Sumber Industri
Malt industri beer
Molasses Carbon Alkohol, butanol, asam asetat
Starch dan dextrin Carbon Fermentasi ethanol
Cellulose Carbon alkohol, nutanol, acetone dan isopropanol
Whey Carbon xanthan gum, 2,3 butadienol, asam lactat
Vegetable oil soy, palm Carbon fermentasi antibiotic
Methanol Carbon asam glutamat, serin dan vitamin B12
Corn steep liqour Nitrogen Asam laktat
Soya meal nitrogen antibiotik
9
-
8/10/2019 2 Medium Dan Kinetik Farmasi
10/32
Design optimasi medium
menggunakan statistik method
Placket- Burman screening design method
Digunakan untuk menyeleksi medium yang paling
berpengaruh dalam pertumbuhan cell
Sehingga dapat diperoleh medium
medium yangpaling dibutuhkan mikroorganisme
Optimasi medium menggunakan respon
surface method
Diperoleh komposisi medium yang digunakanuntuk menghasilkan produk secara optimal
10
-
8/10/2019 2 Medium Dan Kinetik Farmasi
11/32
metode statistik untuk perancangan percobaan, pemodelan matematik, optimasi
dan analisis statistik dalam penelitian persamaan polinomial kuadratik dikembangkan untuk memperkirakan hasilpercobaan sebagai fungsi dari interaksi antara variabel bebas
Type RSM yang dipakai yaitu Central Composite design
Contoh design optimasi pembuatan nata de coco
Design eksperimen Respon Surface method
Variabel bebas
Range dan level
Star point (-) Low level (-1)Center level
(0)
High Level
(+1)
Star point
( + )
Konsentrasi gula (%w/v)
0.41%
1%
10%
20%
21.5%
Lama fermentasi (hari) 4 6 10 14 16
pH 7 6 4.5 3 2
-
8/10/2019 2 Medium Dan Kinetik Farmasi
12/32
Run Konsentrasi
gula
Lama
fermentasi
pH
1 - 1 -1 -1
2 - 1 -1 +
3 -1 +1 -1
4 -1 +1 +1
5 +1 -1 -1
6
+1
-1
+1
7 +1 +1 -1
8 +1 +1 +1
9 -1 0 0
10 +1 0 0
11
0
-
0
12 0 + 0
13 0 0 -
14 0 0 +
15 0 0 0
16
0
0
0
Tempuhan berdasarkan design eksperiment menggunakan Central
Composite Design
-
8/10/2019 2 Medium Dan Kinetik Farmasi
13/32
13
Kinetic of microbial growth for
fermentation product
-
8/10/2019 2 Medium Dan Kinetik Farmasi
14/32
14
What are the value of rates?Rates of consumption or production are obtained from
mass balance over reactors
Mass balance over reactors
Transport + conversion = accumulation
(in out) + (production consumption) = accumulation
Batch: transport in = transport out = 0
Chemostat: accumulation = 0, steady state
Fed batch: transport out = 0
-
8/10/2019 2 Medium Dan Kinetik Farmasi
15/32
Continuous Reactors
Chemostat - CSTR - continuous stirred tank
reactor for the cultivation of cells.
mixing supplied by impellers and rising gas bubbles
assume complete mixing - composition of any phases
do not vary with position
liquid effluent has the same composition as the reactor
contents
-
8/10/2019 2 Medium Dan Kinetik Farmasi
16/32
Chemostat with Recycle
F - nutrient flow rate V - reactor volume
X1- x concentration in reactor
X2- X concentration in effluent
XR- X concentration in recycle
FR- recycle flow rate
F,
X0V,
X1
F,
X2
F+FR,
X1
FR, XR
-
8/10/2019 2 Medium Dan Kinetik Farmasi
17/32
Chemostat with Recycle Cell
mass equation
Acc = in - out + gen
F X0 + FRXR - (F+ FR) X1+ VmX1= V dt
dX1
F, X0V,
X1
F, X2
F+FR,
X1
FR, XR
-
8/10/2019 2 Medium Dan Kinetik Farmasi
18/32
18
How are rates defined?
Rate (ri) = amount i per hour / volume of reactor
Biomass specific rate (qi)
qi =amount per hour / amount of organism in reactor
Thus:
Substrate (-rS) = (-qS)CX
Biomass rX= mCX
Product rP= qPCX
Oxygen (-rO2) = (-qO2)CX
reactorm
hourikg
3
/.
Xkg
hourikg
.
/.
ri= qiCX
-
8/10/2019 2 Medium Dan Kinetik Farmasi
19/32
19
Yield = ratio of rates
Yij= ij
Xi
Xj
i
j
q
q
Cq
Cq
r
r
irate
jrate
.
.
YSX= rate of biomass production / rate of substrate
consumption [g biomass/g substrate]YOX= rate of biomass production / rate of oxygen
consumption [g biomass/g oxygen]
-
8/10/2019 2 Medium Dan Kinetik Farmasi
20/32
20
Introduction
Cell growth and product formation are complex processesreflecting the overall kinetics and stoichiometry of the
thousands of intracellular reactions that can be observed within
a cell.
Thermodynamic limit is important for process optimization.The complexity of the reactions can be represented by a simple
pseudochemical equation.
Several definitions have to be well understood before studying
this chapter, for example: YSXmax, YATP X, YOX, maintenancecoefficient based on substrate (ms).
-
8/10/2019 2 Medium Dan Kinetik Farmasi
21/32
21
Composition of biomass
Molecules
Protein 30-60 %
Carbohydrate 5-30 %
Lipid 5-10 % DNA 1 %
RNA 5-15 %
Ash (P, K+, Mg2+, etc)
Elements
C 40-50 %
H 7-10 %
O 20-30 % N 5-10 %
P 1-3 %
Ash 3-10%
Typical composition biomass formula: C1H1.8O0.5N0.2
-
8/10/2019 2 Medium Dan Kinetik Farmasi
22/32
Figure 6.14
LAG
LOG
STATIONARY
DEATH
SENESCENCELog10of#of
CFU/ml
-
8/10/2019 2 Medium Dan Kinetik Farmasi
23/32
Monod Equation
Growth equation (Monod equation)
m = mmax S/ (Ks +S)
Substrate conc (S)
Specific
growth
rate
-
8/10/2019 2 Medium Dan Kinetik Farmasi
24/32
Stationary Phase
Growth equations with substrate exhaustion:
Cells: (rate of change of cell concentration)dX/dt = mX = mmaxS/(Ks + S) * X
Substrate: (rate of change of substrate conc)
dS/dt = -1/Yx/SdX/dt = -1/Yx/SmmaxS/(Ks + S) X
S b t t d l ti i b t h
-
8/10/2019 2 Medium Dan Kinetik Farmasi
25/32
Substrate depletion in batch
culture
B t h R t Ki ti R l
-
8/10/2019 2 Medium Dan Kinetik Farmasi
26/32
Batch Reactor KineticsReal
data
0.01
0.1
1
10
100
0:00:00 6:00: 00 12:00: 00 18:00: 00 24:00: 00 30: 00:00 36: 00:00
Time (hr:min:sec)
OD
0
0.5
1
1.5
2
2.5
Glucose(g/L)
-
8/10/2019 2 Medium Dan Kinetik Farmasi
27/32
Batch Reactor Kinetics
0.001
0.01
0.1
1
10
100
0 5 10 15 20 25
Time (hr)
OD@60
0nm
-2
0
2
4
6
8
10
12
Glucoseco
nc(g/L)
Series1
Series2
-
8/10/2019 2 Medium Dan Kinetik Farmasi
28/32
Antibiotic production
There are over 10 000 different antibioticsknown, but only about 200 in commercial
use, since most new antibiotics are nobetter than existing ones.
There is a constant search for newantibiotics. Antibiotics are the most-prescribed drugs and are big business.
Finding a new antibiotic and getting it onto the market is a very long process andcan take 15 years.
-
8/10/2019 2 Medium Dan Kinetik Farmasi
29/32
Antibiotic Production Methods
Antibiotics are produced on an industrial scaleusing a variety of fungi and bacteria.
Penicillin is produced by the fungus Penic i l l iumchrysogenum which requires lactose, other
sugars, and a source of nitrogen (in this case ayeast extract) in the medium to grow well.
Like all antibiotics, penicillin is a secondarymetabolite, so is only produced in the
stationary phase. What sort of fermenter does it require?
It requires a batch fermenter, and a fed batchprocess is normally used to prolong the
stationary period and so increase production.
-
8/10/2019 2 Medium Dan Kinetik Farmasi
30/32
-
8/10/2019 2 Medium Dan Kinetik Farmasi
31/32
-
8/10/2019 2 Medium Dan Kinetik Farmasi
32/32
Further reading
G Rao.2007. Introduction to Biochemical
Engineering , Chapter 6 dan seterusnya
32
Terima kasih