2 medium dan kinetik farmasi

8/10/2019 2 Medium Dan Kinetik Farmasi

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Fermentation TechnologyOptimasi medium dan kinetics

MK : Biotechnology

Fakultas Farmasi

Alwani Hamad, ST, MScProgram Studi Teknik Kimia

Fakultas Teknik

Universitas Muhammadiyah Purwokerto


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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


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Quantification of Microbial Rates using

optimum medium


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4

Microbial rates of consumption or production

C, N, P, S source

H2O

H+

O2

heat

product

CO2

biomass
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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)

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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

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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

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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


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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

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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

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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


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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


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Kinetic of microbial growth for

fermentation product


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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


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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


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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


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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


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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


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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]


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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).


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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


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Figure 6.14

LAG

LOG

STATIONARY

DEATH

SENESCENCELog10of#of

CFU/ml


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Monod Equation

Growth equation (Monod equation)

m = mmax S/ (Ks +S)

Substrate conc (S)

Specific

growth

rate


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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


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Substrate depletion in batch

culture

B t h R t Ki ti R l


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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)


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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


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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.


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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.


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Further reading

G Rao.2007. Introduction to Biochemical

Engineering , Chapter 6 dan seterusnya

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2 medium dan kinetik farmasi

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