User:Fermenter User/Notebook/MACG E7T

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Background color codes:  Pastel Green: Major change of setup this time  yellow: Sung-Hye's correction or question to user

Letter color codes: Black: Basic comment Red: Emergency Information (Power shutdown, building access, Manager's absence etc) Blue: Fermentation Setup information Green: Discussion Orange: Fermentation hrs Purple: Induction hrs

MACG E7T

Day -1 (Mar 30 Mon)
 pH sensor: monitor or control pH   dO2 sensor: monitor or control dO2 Condenser: Gas outlet OD probe: monitor cell growth ==> Not used! Replate it with septum Triple: Addition of media, Acid or Base if necessary MeOH sensor: Monitor EtOH (byproduct)  MeOH sensors also detects ethanol! :See reference #6]            3/30/2009 (13:00)  Pour media 3/30/2009 (13:00) pH calibration 3/30/2009 (13:00) Add antifoam 0.5 ml/fermenter 3/30/2009 (15:10) Autoclave bioreactor ==> 3 bioreactors 3 empty bottles/fermenter (no need this time!) Carbon source, Inoculum, Feeding bottle 3/30/2009 (17:00) Start Temp Control

Day 0 (Mar 31 Tue)
3/31/2009 (10:24) Start F1 software 3/31/2009 (10:25) Start F2 software 3/31/2009 (10:27) Start F3 software 3/31/2009 (10:30) Connect Base (400 ml) for Fer#7 3/31/2009 (11:20) Add Carbon source using pump at 30°C 3/31/2009 (11:27) Calibrate dO2 3/31/2009 (11:27) Set dO2 3/31/2009 (11:30) Set pH for Fer#7   3/31/2009 (12:00) Fermentation 00:00 Inoculation 30 ml each using a pump.                no readings taken today

Day 1 (Apr 1 Wed)
           4/1/2009 (7:00) F3: Base (370 ml)                4/1/2009 (13:00)  Fermentation 25:00 F1: OD (41.1) Contamination? (N)white, budding F2: OD (59.4) Contamination? (N)white, budding F3: OD (57.2) Contamination? (N)white, budding   4/1/2009 (13:00)  Fermentation 25:00 Pepstatin added (13:00) F5: Connect Pump to 500 ml of Feeding media (1xYP; 400g/l Gal) Watson pump: 10 rpm (2rpm pump was used, 10rpm = 12.8 ml/h)

Day 2 (Apr 2 Thu)
           4/2/2009 (07:00) F3: Base (370 ml)                4/2/2009 (9:00)  Fermentation 45:00 F5: OD (113.3) Contamination? (N) F5:Induction 20:00 F6: OD (60.6) Contamination? (N) F6:Induction 00:00 F7: OD (57.8) Contamination? (N) F7:Induction 00:00 <Isis>               4/2/2009 (10:00)  Fermentation 46:00 F5: OD (113.3) Contamination? (N) F5:Induction 21:00 HARVEST <Sung-Hye and Isis>  4/2/2009 (10:30)  Fermentation 45:30 Pepstatin added (00:00) F6: Connect Pump to 250 ml of Feeding media (1xYP; 400g/l Gal) <font style=background-color:yellow>Watson pump: 10 rpm (2rpm pump was used, 10rpm = 12.8 ml/h) <Sung-Hye and Isis>  4/2/2009 (10:30)  Fermentation 45:30 Pepstatin added (00:00) F7: Connect Pump to 250 ml of Feeding media (1xYP; 400g/l Gal) <font style=background-color:yellow>Watson pump: 10 rpm (2rpm pump was used, 10rpm = 12.8 ml/h) <Isis>               4/2/2009 (12:30)  Fermentation 47:30 F6: OD (73.2) Contamination? (N) F6:Induction 2:00, 02 stopped as cells growing too fast F7: OD (68.8) Contamination? (N) F7:Induction 2:00 02 stopped as cells growing too fast, pH was dropping, stopped at 6.0 by Base. <Isis>               4/2/2009 (14:00)  Fermentation 49:00 F6: OD (86.3) Contamination? (N) F6:Induction 3:50 pH 6.5 F7: OD (73.9) Contamination? (N) F7:Induction 3:50 pH 6.6 <Isis>               4/2/2009 (16:00)  Fermentation 51:00 F6: OD (94.5) Contamination? (N) F6:Induction 5:50 F7: OD (84.2) Contamination? (N) F7:Induction 5:50 <Isis>               4/2/2009 (18:00)  Fermentation 53:00 F6: OD (99.4) Contamination? (N) F6:Induction 7:50 F7: OD (82.2) Contamination? (N) F7:Induction 7:50



Day 3 (Apr 3 Fri)
<Sung-Hye>           4/3/2009 (07:00) F3: Base (340 ml) <Isis>               4/3/2009 (07:30) Fermentation 67:30 F6: OD (124.4) F6:Induction 21:30 Contamination?(N)no budding, multiple vacuoles, cells cream color still F7: OD (111.5) F3:Induction 21:30 Contamination? (N) cells cream color still isoamyl acetate smell <Sung-Hye>           4/3/2009 (08:00) Fermentation 68:00 Harvest