Comparison of Growth Medium Bioconversion into Vanda tricolor Biomass in Thin Layer Liquid

Medium Culture System and TIS RITA ® Bioreactor

Annisa Ratna Nurillah, Rizkita Rachmi Esyanti, Robert Manurung

School of Life Sciences and Technology, Institut Teknologi Bandung, Indonesia

annisa.ratna@students.itb.ac.id; rizkita@sith.itb.ac.id; manurung@sith.itb.ac.id

Scope of Problem• In nature, Vanda tricolor take long period to grow time inefficient

• In vitro system: – provide sufficient culture condition– easy to control– homogenous nutrition availability – manipulation possibility

• Problem: which in vitro culture method that could give most favorable condition for biomass to grow?– continuous immersion system (Thin layer medium culture)– temporary immersion system (TIS RITA)

productivity can be enhanced

MethodologyMedium Preparation

MS half strengthcoconut watersucrose cefotaxime

Plantlet & medium analysis

Data analysis: Growth kinetics Sucrose

consumption

Subculture of Vanda tricolor to RITA Bioreactor

Shaken at 100 rpm

10 min immersion time

/12 -hr

5 min immersion time

/12-hr

Incubation condition:Light 12 hr/day25 ⁰C

Incubation

Incubation

Incubation

Result• Plantlet and medium analysis

Culture System

Growth Rate(g/day)

Shoots Elongation Rate

(cm/ day)

Total Sugar Consumption(g)

Conductivity Reduction(mS)

RITA 1 0.013 0.003 7.8 0.66

RITA 2 0.009 0.006 2.2 1.44

Flask 0.012 0.005 14.4 6.775

RITA 1 RITA 2 Flask02468

10121416

Growth Rate (mg/day)Elongation Rate (mm/day)Sugar Consumption (g)Conductivity Reduc-tion (mS)

Hypothetical & Actual Biomass

• Based on Mass & Energy Balance Equations (Respiration)

Culture System

Total Sugar Consumption(g)

Hypothetical Biomass (g)

Actual Biomass (g)

RITA 1 7.8 0.23 0.32

RITA 2 2.2 0.07 0.28

Flask 14.4 0.44 0.17

0.39C12H22O11 + 0.23NH4NO3 + 3.43O2 CH1.27O0.43N0.45 + 4.07H2O + 3.64CO2

Sucrose (C-source) N-Source Oxygen Biomass WaterCarbon dioxide

Inefficiency of sugar consumption

Bioconversion Period• Models were based on equations (Shuler & Kargi, 2002):

a. Growth kinetics:μ = μmax * ([C12H22O11]/ (k C12H22O11 + C12H22O11))

b. Sucrose consumption d/dt [C12H22O11] = - qC12H22O11 X

qC12H22O11 = μ / Yx/C12H22O11 , Yx/C12H22O11 = ΔX / ΔC12H22O11

flask RITA 1 RITA 2X : Biomass production suk : Sucrose consumption

Conclusion• TIS RITA bioreactor was potential to be developed as a

better method for shoot production than thin layer liquid medium culture.

• The period of culture immersion directly affected the growth of orchids. – Five minutes immersion in medium was an optimal

immersion period for growth of the Vanda tricolor shoots compared with that on 10 minutes in RITA bioreactor.

– Growth rate of culture in RITA bioreactor system with optimal immersion period (5 minutes) was higher compared to the thin-layer liquid medium culture system.

Thank You