User:Fermenter User/Notebook/BROM A18

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BROM A18


Day -2 (Nov 22 Mon):

      <Dennis>           11/22/2010 ()
                               Thaw glycerol stock and grow in 5-ml MD media (prepared in summertime)
                               Incubator in Fermentation Suite used
      <Dennis>           11/22/2010 (before go home)
                              Preparing 5-L of Basal Medium (Defined media)
                              Ortho-phosphoric acid (85%) was ordered
                              PTM1 Salt is form BROM-A16

Day -1 (Nov 23 Tue): MEDIA PREPARATION DAY

      <Sung-Hye>         11/23/2010 (14:00)    Assemble bioreactors:   
                              1. Calibrate pH sensors                                         
                              2. 5.0-L of Basal Salts (w/o Glycerol):  
                              3. 1-ml Antifoam 
<Sung-Hye> 11/23/2010 (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> 11/23/2010 (14:30) Autoclave <Sung-Hye> 11/23/2010 (16:00) Autoclave completed! <Dennis> 11/23/2010 (11:50 - 12:00) Transfer cells to fresh 55-ml BMGY media Incubator in Fermentation Suite used

Day 0 (Nov 24 Wed): INOCULATION DAY

      <Sung-Hye>         11/24/2010 (11:00)   Set temperature to 28.5°C 
      <Sung-Hye>         11/24/2010 (11:00)   Purge Air 1L/min at 900 rpm 
      <Sung-Hye>         11/24/2010 (11:30)     Add 22 ml of PTM1 Salt/bioreactor
                                                Add 158 ml of 50% Glycerol
      <Sung-Hye>         11/24/2010 (11:00)   Adjust pH to 5.0 with 140 ml NH4OH 
      <Sung-Hye>         11/24/2010 (11:00)      Start pH control 
                                                  F1: No Acid Base, 1M (< 400 ml)
      <Sung-Hye>         11/24/2010 (11:00)      Start Fermenter #1 software 
                         11/24/2010 (11:00)      Calibrate dO2 probes 
                         11/24/2010 (11:00)      Strt dO2 control 
<Dennis> 11/24/2010 (13:30) Check contamination of starter culture
<Dennis&Sung-Hye> 11/24/2010 (14:30) Fermentation 0:00 Flask (from streak stock) OD: 10 Inoculate 45 ml/bioreactor

Day 1 (Nov 25 Thu):

      <Dennis>           11/25/2010 (14:30)      Fermentation 24:00  
                            F1 Base(300 ml) OD(11.3) Contamination(N) Pressure(N) Change Filter(N)

Day 2 (Nov 26 Fri) MeOH Induction Day 1

      <Dennis>           11/26/2010 (12:00)      Fermentation 46:00  
                            F1 Base(210 ml) OD(17.25) Contamination(N) Pressure(N) Change Filter(N)
      <Sung-Hye>         11/26/2010 (12:30 - 15:30)  Fermentation 48:00  
                            F1 Fed-Batch
      <Dennis>           11/26/2010 (15:00)      Fermentation 48:00  MeOH induction 00:00  
                            F1  OD (19.05)   
      <Sung-Hye>         11/26/2010 (15:30)      Fermentation 48:00  MeOH induction 00:00 
                            Calibrate MeOH sensor  6850 mV = 0.5%
                            Start MeOH induction
      <Sung-Hye>         11/26/2010 (15:30)      Fermentation 48:00  MeOH induction 00:00 
                            Refill Base (150 ml + 250 ml = 400 ml)

Day 3 (Nov 27 Sat): MeOH Induction Day 2

      <Dennis>           11/27/2010 (14:00)      Fermentation 96:00  MeOH induction 24:00  
                            F1: Base(250 ml) MeOH(300 ml) OD(39.8) Contamination(N) Pressure(N) Change Filter(N)
                                Activity assay (9.79 units/100ul), (2.29 mg/5-L)

Day 4 (Nov 28 Sun): MeOH Induction Day 3

      <Dennis>           11/28/2010 (14:00)      Fermentation 120:00  MeOH induction 48:00  
                            F1: Base(240 ml) MeOH(270 ml) OD(35.05) Contamination(N) Pressure(N) Change Filter(N)
                                Activity assay (21.28 units/100ul), (4.98 mg/5-L)

Day 5 (Nov 29 Mon): MeOH Induction Day 4

      <Dennis>           11/29/2010 (14:00)      Fermentation 144:00  MeOH induction 72:00  
                            F1: Base(240 ml) MeOH(250 ml) OD(37.8) Contamination(N) Pressure(N) Change Filter(N)
                                Activity assay (22.59 units/100ul), (5.29 mg/5-L)

Day 6 (Nov 30 Tue): MeOH Induction Day 5

      <Dennis>           11/30/2010 (8:30)      Fermentation 168:00  MeOH induction 96:00  
                            F1: Base(240 ml) MeOH(245 ml) OD(37.8) Contamination(N) Pressure(N) Change Filter(N)
                                Activity assay (25.58 units/100ul), (5.99 mg/5-L)
      <Dennis>           11/30/2010 (11:30)      Fermentation 168:00  MeOH induction 96:00  
                            F1: Base(240 ml) MeOH(245 ml) OD(37.8) Contamination(N) Pressure(N) Change Filter(N)
                                Activity assay (24.71 units/100ul), (5.78 mg/5-L)
      <Sung-Hye>         HARVEST




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