This document discusses a study on the effect of scaling up Eurycoma longifolia cell suspension cultures in a bioreactor and treating with UV light on growth and secondary metabolite production. The study found that (1) E. longifolia cell cultures grown in a bioreactor had a faster growth rate than in flask culture, (2) UV light inhibited the growth of E. longifolia cell culture, and (3) elicitation with UV light increased the production of secondary metabolites in both flask and bioreactor scale cultures.

1. Januar Natanael, Rizkita Rachmi Esyanti,Robert Manurung
Department of Biological Engineering
School of Life Sciences and Technology
Institut Teknologi Bandung, Indonesia

2. 
 Eurycoma longifolia Jack or tongkat ali can be used as a
medicine to cure malaria, anti-inflammatory, anti-
microbial, aphrodisiac and is still being developed as
anticancer (Bhat, et al., 2010)
Problem:
 excess exploitation of this plants  critically endangered
(Sudiarto, et al., 2002)
Alternative method  tissue culture technique
Introduction

3. 
 Elicitation is a treatment given to culture, such as by
exposing ultra-violet light, to induce the production
of specific metabolites
 UV elicitation has been proved able to induce the
formation of the enzyme tryptophan decarboxylase
on Catharantus roseus L.  alkaloid production
increased (Ouwerkerk et al., 1999)
Introduction

4. 
 Biomass propagation in the bioreactor is a way to
increase the biomass and secondary metabolites
production in a larger scale.
 There are several factors that must be considered in
the scale-up process, such as: profiles of stirring, the
solubility of oxygen in the medium, the exact
dimensions of the reactor, the size of the cells used,
and many more (Taticek et al., 1991)
Introduction

5. 
 In this study, the effect of scale-up of cell suspension
cultures of E. longifolia in bioreactor conditions and
treated by UV light on the growth and secondary
metabolite production will be evaluated.
Objective

6. 
Methods
Subcultivated
callus in fresh
solid media
Initiation
cell
suspension
from callus
Next
MS Medium half strength
1,1 ppm 2,4-D and 1 ppm kinetin
MS Solid Medium half stregth
2,2 ppm 2,4-D and 2 ppm kinetin

7. 
Methods
Subcultured cell
aggregate into
fresh liquid and
bioreactor for
treatment
Analysis
Treatment:
Exposed by UV 18
hr/day
Rotation: 100 rpm
Air flow rate: 0.3 vvm

8. 
E. longifolia Cell Culture
Specific Growth Rate
y = 0.0257x + 0.0468
R² = 0.9129
y = 0.0225x - 0.019
R² = 0.9316
y = 0.0549x
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
0 2 4 6 8 10 12 14
ln(Xt/Xo)
Day
Flask-Control
Flask-UV
Bioreactor-UV
*
* Ouwerkerk et al., 1999 dan Stephanopoulus et al., 1998).
** Shuler dan Kargi, 2002 dan Schlatmann et al., 1994
**

9. 
Medium Conductivity Profile
of E. longifolia Cell Culture
2
2.5
3
3.5
4
4.5
5
5.5
6
0 2 4 6 8 10 12 14
Conductivity(mS)
Day
Flask-Control
Flask-UV
Bioreactor-UV
*Taya et al., 1989 dan Hahlbrock, 1975

10. 
Sucrose Consumption of
E. longifolia Cell Culture
y = -0.56x + 24.86
R² = 0.99145
y = -0.43x + 25.34
R² = 0.91009
y = -0.18x + 25
13.00
15.00
17.00
19.00
21.00
23.00
25.00
27.00
-1 4 9 14
SucroseLevel(g/L)
Day
Flask-Control
Flask-UV
Bioreactor-UV
* Shuler dan Kargi, 2002
** Shuler dan Kargi, 2002
**
*

11. 
Secondary Metabolites of
E. longifolia Cell Culture
Day
Metabolites in Control
Culture
Amount Day
Metabolites in UV-Exposed
Culture
Amount Day
Metabolites in UV-
Exposed Bioreactor
Amount
9 Eurylactone 3.75% 4 Squalene 4.38% 14 Canthin-6-one 6.81%
12
5-(hydroxymethyl)-2-
Furancarboxaldehyde
20.52% 9 Squalene 8.04% 14 Beta-carboline 29.24%
14 Canthin-6-one 2.67% 12 Quasinoid 4.03%
14 Squalene 3.50% 14 Canthin-6-one 5.12%
14 Beta-carboline 25.11%
* Arvy et al., 1994; Fett-Neto dan Zhang, 1994; dan Ouwerkerk et al., 1999

12. 
 E. longifolia cell cultures that were grown in
bioreactor had a faster growth rate than in the flask
culture.
 UV light inhibits the growth of E. longifolia cell
culture
 The faster the rate of biomass formation, the faster
the rate of sucrose and mineral salts consumption.
 Elicitation with UV light increases the production of
secondary metabolites in both the flask scale culture
and bioreactor scale culture.
Conclusions

13. 
Thank You