Task 3By: Erdenesuvd Bat-Erdene

25,000L PBR

3d PBR Pond

M2.5 Supernatant

E-2

M3.5 Supernatant

E-3

E-1

M2.1 CO2

M7.3 Hot Air

M7.1 EvaporatedWaste Water

6000L PBRE-2 E-4E-3

E-8E-7E-6

PulveriserSpray DryerCentrifuge

M1 INOCULUM

M2.3 Medium

M2.2 AIR

M2 Cell Culture

M7 Algae Paste

M6 Sedimented

Slurry

M5 Cell Slurry

M4 Cell Culture

M3 Cell Culture

M8 Dried Algae

M1.1 CO2M1.3

Medium

M1.2 AIR

M3.1 CO2M3.3

Medium

M3.2 AIR

M4.1 CO2M4.2

Medium

M4.4 Supernatant

E-4

M7.2 Waste Air

M6.1 Supernatant

M1.4Waste Gas

M3.4Waste Gas

M2.4Waste Gas

M8.1 Biomass Waste

E-9

ExtractionM9

Sedimentation Tank

E-5

M5.1 Supernatant

E-10

Purification

M9.1 Biomass Waste

M10

M10.1 BiomassWaste

M10.2 Astaxanthin

M4.3Waste Gas

M7.4 Biomass Waste

Overall Process of 5 tons/year Astaxanthin production plant

• Biomass concentrations in Inflow and Outflow Basis:• Based on residence time 5 days and daily 6 hours of solar illumination in Kunming, China

(Ref 1).

• Method: biomass dry weight 0.0004 kg/L in the final pond culture

• In the ponds the cell culture is harvested but not grown. So the cell concentration in the pond will be 6x10^7cells/L

• Per cell biomass: = 0.0004/(6x10^7) = 6.67x10^-12 kg/cell, confirms with the 1x10^-12kg/cell from Ref 2.

• Biomass IN (kg/day) = Start cell biomass (cells/L) x per cell biomass (kg/cell) x Volumetric flowrate (L/day)

• Biomass OUT (kg/day) = End cell biomass x per cell biomass x Volumetric Flow rate

Start Cell biomass End cell biomass

6 x 10^7cells/L 5 x 10^8 cells/L

Mass Balance of Dry Biomass

• Nutrient medium: 10 mM KNO3, 2 mM Na2HPO4, 0.5 mM CaCl2, 0.5 mM MgSO4, 2 mM NaHCO3 (Ref 1)

• Air: Aeration rate 0.05 vvm (Ref 1)• Mass concentrations of components in Air: 23.2%

O2, 75.5% N2, 1.3% Ar. Assuming Oxygen is used fully, N2 and Ar are inert. Oxygen concentration 150% saturation is desired. (Ref 2).

• CO2 in: 15vol% going into 6000L and 25,000L PBRs (Ref 1)

• CO2 out: 75mass% going out from PBRs (Ref 3).

Mass Balance

E-16000L PBR

M1 INOCULUM M2 Cell Culture

M1.1 CO2

M1.3 Medium

M1.2 AIR

M1.4Waste Gas

Dry Biomass 1.38 kgKNO3 0.42 kgNa2HPO4 0.12 kgCaCl2 0.023 kgMgSO4 0.025 kgNaHCO3 0.070 kgWater 412.68 kg

CO2 5.18 kg

Oxygen 73.63 kgNitrogen 239.61 kgArgon 4.13 kg

KNO3 3.49 kgNa2HPO4 0.98 kgCaCl2 0.19 kgMgSO4 0.21 kgNaHCO3 0.58 kgWater 2960.90 kg CO2 3.89 kg

Nitrogen 239.61 kgArgon 4.13 kg

Dry Biomass 11.53 kgWater 3444.47 kg

Total mass going In = 3703.62 kg/dayTotal mass going Out = 3703.62 kg/dayResidence time 5 days

0 1 2 3 4 5 6 70

100000000

200000000

300000000

400000000

500000000

600000000

f(x) = 88000000 x + 60000000

Biomass Growth

Days

Biom

ass c

ells/

L

MASS BALANCES kg/day

25,000L

PBR

M2.1 CO2

E-2

M2.3 Medium

M2 Cell Culture M3 Cell Culture

M2.4Waste Gas

M2.2 AIR

CO2 43.2 kg

Oxygen 153.39 kgNitrogen 499.18 kgArgon 8.60 kg

KNO3 29.12 kgNa2HPO4 8.18 kgCaCl2 1.60 kgMgSO4 1.73 kgNaHCO3 4.84 kgWater 25134.34 kg

CO2 32.40 kgNitrogen 499.18 kgArgon 8.60 kg

Dry Biomass 11.53 kgWater 3444.47 kg

Dry Biomass 96.05 kgWater 28703.95 kg

Total mass going In = 29340.18 kg/dayTotal mass going Out = 29340.18 kg/dayResidence time: 5 days

MASS BALANCES kg/day

ENERGY BALANCES

Energy Balance for 6000L PBRs (Assume Turbulence power and solar radiation negligible at this stage)Cooling Duty = Energy In – Out + Turbulence Power + Solar Radiation

Air Supply (O2,N2,Ar) CO2 Supply Cell culture Medium and

Water Total

m (Kg/Day) 317.37 5.18 414.72 2966.35Cp (KJ/Kg/C) 1.005 0.8439 4.184 4.184Inlet Temperature 25C 25C 20C 25COutlet Temperature 20C 20C 20C 20C∆T 5 5 0 5Q (KJ/day) 1594.78 21.86 0 62056.04

Q(KW) 0.0185 0.000025 0 0.718 0.737 kW

• Energy Balance for 25,000L PBRs: • Cooling Duty = Energy In – Out + Turbulence Power + Solar Radiation

ENERGY BALANCES

Air Supply (O2,N2,Ar) CO2 Supply Cell culture Medium and

Water Total

m (Kg/Day) 661.17 43.2 3456 25179.87Cp (KJ/Kg/C) 1.005 0.8439 4.184 4.184Inlet Temperature 25C 25C 20C 25COutlet Temperature 20C 20C 20C 20C∆T 5 5 0 5Q (KJ/day) 3322.38 182.28 0 526762.88

Q(KW) 0.0385 0.00211 0 6.0968 6.137 kW

Photo bioreactor process control parameters Values RangesBioreactor culture temperature Below 25 C 10-25 CBioreactor aeration rate 0.05v/v/mpH 6.5-7Starting cell concentration of PBRs 6 x 10^7 cells/l 5-8 x 10^7 cells/lFinal cell concentration for PBRs 5 x 10^8 cells/l 5-7 x 10^8 cells/lPhoto bioreactor process performance parameters Estimated values RangesPower consumption for cooling a 6000L PBR 60 kWh 0-120 kWhPower consumption for cooling a 25000L PBR 160 kWh 0-320 kWhPower consumption for turbulence a 6000L PBR 15 kWPower consumption for turbulence a 25000L PBR 67.5 kWAeration power input for the 6000L bioreactors 0.6 kWhAeration power input for the 25000L bioreactors 2.5 kWhDays for 6000L bioreactor cell culture to be ready 5 days 4-6 daysDays for 25000L bioreactor cell culture to be ready 5 days 4-6 days

Summary of Operational Parameters

6000L PBRs 5days = V / (3703.62 kg/day)V = 18518.1 kg 3 x 6000L PBRs needed

25,000L PBRs5days = V / (29340.18 kg/day)V = 146700.9 kg/ (1000kg/m3) = 146.70 m36 x 25,000L PBR needed

Mechanical Design for:

Equipment Volumes

Choice of photobioreactor

• If one desires to provide large quantities of cheap/free light to the cultures, the cultures need to be taken outside.

• To be free from contamination they should be enclosed.

• Each 25,000L PBR module has 100m2 land surface area exposed to the sun

• Disadvantage: Large area needed (6x25000LPBRs need area of 780m2)

Mechanical design of 25,000L PBR

• 4 parallel plastic tubes each 0.41 m diameter 34.5 m length laid on an impermeable surface (Ref 2).

• Turbulence: air lift pump, Re ~ 4000• Air lift pump Duty: 2–3.4 kW m-3 • Cooler: To control temperature 15 to 25C. PBR

is automatically flooded with cold sea water from Kunming sea 600m under ocean surface.

• Cooler duty: 160 kWh

Mechanical design of 25,000L PBR

D= 0.41m

L= 34.5m

W = 3.89 m

X 6

2D Model of 25,000L PBR

In real life…

3D Model of 25,000L PBR

• Pumps• Air lift pump for cell culture going into PBRs• Centrifugal pump for outlet to the next PBR• Cascade Pumping System for cool water pump from deep ocean under 600m

deep to the cooling pool.• Air Compressor Duty: 1.25kW x 6• Filters• Air and CO2 filters: 0.2micrometre membrane filter• Medium filters:2micrometre membrane filter• Pipes and tubes: 0.41 m diameter 34.5 m length PVC Plastic Pipes, narrow PVC

tubes used for sparging CO2 and Air to PBRs• Line sizes for in and outflow of cell culture and medium: Nominal Size 2.5, OD

73 mm, Sch 40• Valves: Diaphragm valve for air, CO2 and water (fail-closed)

– Gate valve for cell culture• Tanks• CO2 Tank: Compressed tank stainless steel• Medium Tank: Stainless steel Tank

Specifications of Ancillary Items for 25,000L PBR

• An outline of the intended control system:

• Computer controlled parameters:• Nutrient concentration, Dilution rate,

Temperature, pH, Turbulence, Growth rate• Cell count system: Model Z1 Coulter Counter

Ref 2.

Control System Specification

P&IDP&ID

1. Make sure all valves closed V-9, V-8, V-13, and V-102. Medium Supply valve is opened, V-103. The plastic tubes were filled with medium 4. Medium discharge V-11, V-12 is opened and medium pump E-4 started5. Medium drained to Supernatant storage6. Close V-10 Medium supply7. Close Drain valve V-128. Deep sea water V17 and V-18 opened and Pump E-6 was started9. Switch isolators: cell culture V-6, V-8 and E-3, CO2 supply V-1 and V-9,

medium supply V-10, Air supply V-4, V-16 valves were opened10. Conditions of supplies are fully computer controlled11. After the tube volume is filled, all incoming valves V-8, V-9, V-10, V-13

etc. are closed12. After 5 days V-13, V-14 opened and pump E-5 started to transfer to the

next PBRs

Start Up procedure of 25,000L PBR

1. All incoming valves and pumps to PBR is closed.

2. H2O cooler and all isolators switched off3. Close V-13, pump E-5, V-14 valve4. Open drainage valve fully V-125. PBR system was emptied of medium to

supernatant waste storage

Shutdown Procedure

• Ref 1: Jian Li, Daling Zhu (2011) An Economic assessment of astaxanthin production by large scale cultivation of HP

• Ref 2: Miguel Olaizola (2000) Commercial production of astaxanthin from HP using 25,000L outdoor PBR

• Ref 3: Shu KI Tsang (2004) Optimal Harvesting strategy for HP using stella based model

• Ref 4: http://www.tatup-journal.de/downloads/2012/tatup121_noua12a.pdf

• Ref 5: PBRs design and performance with respect to light and energy input Otto, Pulz (1998)

References

• Any questions?

Thank you for you attention