User:Fermenter User/Notebook/BROM A16

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BROM A16


Day -2 (Nov 8 Mon):

      <Dennis>           11/8/2010 ()
                               Thaw glycerol stock and grow in 5-ml MD media (prepared in summertime)
                               Incubator in Fermentation Suite used
      <Sung-Hye>         11/8/2010 (before go home)
                              Preparing 5-L of Basal Medium (Defined media) <-- Phosphoric Acid used
                              Preparing 1-L of PTM1 salt
Freshly filtered PTM salt
Defined Media for 5-L

Day -1 (Nov 9 Tue): MEDIA PREPARATION DAY

      <Sung-Hye>         11/9/2010 (14:00)    Assemble bioreactors:   
                              1. Calibrate pH sensors                                         
                              2. 5.0-L of Basal Salts <-- Phosphoric Acid used  
                              3. 1-ml Antifoam 
<Sung-Hye> 11/9/2010 (14:00) Prepare bottles 1) no Acid need 2) Base, 500-ml (tubing connected) x2 3) MeOH, 1-L (tubing connected) x2 4) Additives/Inoculum, 1 x 500-ml (tubing connected) x2 5) Water, 2-L

<Sung-Hye> 11/9/2010 (15:22) Autoclave <Sung-Hye> 11/9/2010 (17:00) Autoclave completed! <Sung-Hye> 11/9/2010 (17:00) Set temperature to 28.5°C <Sung-Hye> 11/9/2010 (17:00) Purge Air 1L/min at 900 rpm <Sung-Hye> 11/9/2010 (17:30) Adjust pH to 5.0 with 140 ml NH4OH <Dennis> 11/9/2010 (11:50 - 12:00) Transfer cells to 110-ml BMGY media Incubator in Fermentation Suite used
pH Adjustment with NH4OH, Fermenter #1
pH Adjustment with NH4OH, Fermenter #2

Day 0 (Nov 10 Wed): INOCULATION DAY

      <Sung-Hye>         11/10/2010 (10:18)      Start Fermenter #1 software 
                         11/10/2010 (10:20)      Start Fermenter #2 software 
                         11/10/2010 (11:00)      Calibrate dO2 probes 
                         11/10/2010 (11:00)      Strt dO2 control 
<Dennis> 11/10/2010 (11:30) Check contamination of starter culture
<Sung-Hye> 11/10/2010 (16:30) Add 22 ml of PTM1 Salt/bioreactor <Dennis&Sung-Hye> 11/10/2010 (12:00) Fermentation 0:00 Flask 1 (from frozen stock) OD: 15.3 Flask 2 (from streak on MD plate) OD: 27.8 Inoculate 45 ml/bioreactor <Sung-Hye> 11/10/2010 (12:00) Start pH control F1: No Acid Base (400 ml) F2: No Acid Base (400 ml)

Day 1 (Nov 11 Thu):

      <Dennis>         11/11/2010 (12:00)      Fermentation 24:00  
                            F1 (Frozen Stock) Base(400 ml) OD(-) 
                            Contamination(N but strangely shaped translucent structures were found) 
                                Pressure(N) Change Filter(N)
                            F2 (from Streak)  Base(400 ml) OD(-) 
                            Contamination(N but strangely shaped translucent structures were found) 
                                Pressure(N) Change Filter(N)

Day 2 (Nov 12 Fri)

      <Dennis>         11/12/2010 (12:00)      Fermentation 48:00  
                            F1 Base(400 ml) OD(1.1) Contamination(N) Pressure(N) Change Filter(N)
                            F2 Base(400 ml) OD(2.25) Contamination(N) Pressure(N) Change Filter(N)


Day 3 (Nov 13 Sat)

      <Dennis>           11/13/2010 (12:00)      Fermentation 72:00  MeOH induction 24:00  
                            F1: Base(400 ml) OD(2.1) Contamination(N) Pressure(N) Change Filter(N) 
                            F2: Base(400 ml) OD(3) Contamination(N) Pressure(N) Change Filter(N)

After 3 days, we decided to end the fermentation due to lack of yeast growth.

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