Lactic Acid Production

UNIVERSITI KUALA LUMPUR MALAYSIAN INSTITUTE OF CHEMICAL AND BIOENGINEERING TECHNOLOGY 

FERMENTATION TECHNOLOGY (CBB20203 )

MINI PROJECT

PRODUCTION OF LACTIC ACID (MATERIAL & PROCEDURE)

GROUP

: MOHD SALEHAN BIN HASSAN

55202210266

AHMAD SHAFIQ BIN AHMAD FUAD

55202210134

MOHAMAD NORAZWAN BIN BAHARIN

55202210381

QAIYYUM AZIZI BIN MOHD NOOR

55202210221

SECTION

: 3 BCB EX

LECTURER 

: SHARIFAH SOPLAH BT. SYED ABDULLAH

MATERIAL: A) Bacteria:

y

 Lactobacillus

sp.

B) Chemical:

y

Glucose.

y

Yeast

y

Ammonium

y

Manganese

y

Cor n

y

Distilled

y

y

extract. hydr ogen phosphate, (NH4)2HPO4

sulf ate,

MnSO4

steep liquor. water.

 Natr ium hydr oxide, NaOH. Hydr ochlor ic acid, HCL.

C) Apparatus:

y

2 L Bioreactor 

y

Incubator

y

50 ml

y

250 ml Er lenmeyer flask 

y

Loo p

y

1l

y

1L Measur ing

shaker 

Er lenmeyer flask 

Beaker  cylinder 

METHODS: A) Inoculum:

y

Prepare 30 g/L of glucose, 10 g/L of yeast extract, 2g/L of (NH4)2HPO4, and 0.1 g/L of  MnSO4 in 1L of distilled water.

y

Adjust

culture medium to pH 6.0.

y

Dispense 15 mL

y

Transf er

in a 50

mL

Er lenmeyer flask and autoclave for 15 min at 121OC.

a loo pful of  Lactobacillus culture f r o m agar slant into the first

medium

and incubate for 12-14 h at 36 °C in incubator shaker at 200 rpm. y

Prepare another f er mentation medium of volume 100 mL in 250

mL

and

O

autoclave for 15 min at 121 y

Transf er

C.

2 ml of the inoculum to the flask and incubate for 12-14 h to reach the

exponential gr owth at shak ing 200 rpm at 36 °C.

B) Fermentation:

y

Prepare 30 g/L of cor n steep liquor, 75 g/L of glucose, 1.5 g/L of yeast extract, 2g/L of (NH4)2HPO4, and 0.1 g/L of  MnSO4 in 1L of distilled water.

y

Adjust

f er mentation medium to pH 6.0.

y

A

y

Transf er 1.35 L

2 L tank  bioreactor is used for this f er mentation. of f er mentation medium into  bioreactor and ready to autoclave O

for 15 min at 121 y

Transf er 150 ml

y

Dur ing

C.

of innoculum into  bioreactor.

all f er mentation, agitation speed is fixed at 200 rpm and the tem perature

within the f er menter was contr olled at 36 °C and pH at 6.0. y

Calibrate

y

The

the pum p rate before starting any continuous f er mentation.

sam ples were removed aseptically at regular intervals every 4 hours until 24

hours for further analyses.

C) Analysis y

Lactic

acid was quantified  by HPLC analysis cou pled with a UV var ia ble

wavelength detector set to 210 nm. y

An Aminex

HPX-87H ion-exclusion column was eluted with 5 mM sulfur ic acid

solution at 0.6 mL/min, and the column te m perature was maintained at 35 °C. y

Cell

gr owth was deter mined tur  bidimetr ically by a spectr o photometer at 660 nm,

and the values thus obtained were then converted to dry cell mass via calculation with the appr o pr iate standard curve.