User:Fermenter User/Notebook/MACG F2

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<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext"></div><div style="display:none;" id="page">User:Fermenter User/Notebook/MACG F2</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

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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 F2

Day -2 (Feb 15 Sun)

   <Ann>                 2/15/2009 (13:45) Prepare for small culture (15mL).

Day -1 (Feb 16 Mon)

  <Bioreactor headplate configuation>
   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)
   
   <Sung-Hye's Note>
    MeOH sensors also detects ethanol! :See reference #1 on the bottom of the page
   
 
   <Sung-Hye>            2/16/2009 (10:30)  Pour media
                         2/16/2009 (11:00)  pH calibration
                         2/16/2009 (11:42)  Antifoam added
                         2/16/2009 (12:36)  Autoclave bioreactor
                                            2 empty bottles/fermenter
                                            Carbon source+Inoculation

2/16/2009 (14:15) Start Temp Control 2/16/2009 (15:00) Add carbon source using pumps at 30°C 2/16/2009 (15:12) F1: Connect Base (400 ml) Start pH control 2/16/2009 (15:17) Start F1 software 2/16/2009 (15:18) Start F3 software
   <Ann>                 2/16/2009 (15:45)  Prepare small culture for inoculum. (215mL)

Day 0 (Feb 17 Tue)

   
    Sung-Hye: CHBE Class Auditing 9:30 - 11:00
   
   <Sung-Hye>           2/17/2009 (7:35)  Connect MeOH sensor
                        2/17/2009 (7:45)  Connect Acid
                        2/17/2009 (7:50)  Calibrate dO2
                        2/17/2009 (7:50)  Set dO2

<Sung-Hye and Ann> 2/17/2009 (8:40) Fermentation 00:00 Inoculation 50 ml each using pump. <Ann> 2/17/2009 (10:00) Fermentation 01:20 F1: OD (0.280) Contamination? (N) F3: OD (0.335) Contamination? (N) <Ann> 2/17/2009 (10:40) Fermentation 02:00 F1: OD (0.300) Contamination? (N) F3: OD (0.380) Contamination? (N) <Ann> 2/17/2009 (11:15) Fermentation 02:35 F1: OD (0.310) Contamination? (N) F3: OD (0.370) Contamination? (N) <Ann> 2/17/2009 (11:50) Fermentation 03:10 F1: OD (0.390) Contamination? (N) F3: OD (0.55) Contamination? (N) Induced with Galactose (55 mL) F3: Induction 00:00 <Ann> 2/17/2009 (12:50) Fermentation 04:10 F1: OD (0.390) Contamination? (N) F3: OD (0.470) Contamination? (N) F3: Induction 01:00 <Ann> 2/17/2009 (13:50) Fermentation 05:10 F1: OD (0.510) Contamination? (N) F3: OD (0.480) Contamination? (N) F3: Induction 02:00 <Ann> 2/17/2009 (15:50) Fermentation 07:10 F1: OD (0.590) Contamination? (N) F3: OD (0.500) Contamination? (N) F3: Induction 04:00 <Ann> 2/17/2009 (17:50) Fermentation 09:10 F3: OD (1.09) Contamination? (N) F3: Induction 06:00 <Ann> 2/17/2009 (18:50) Fermentation 10:10 F1: OD (0.950) Contamination? (N) <Ann> 2/17/2009 (19:50) Fermentation 11:10 F3: OD (1.780) Contamination? (N) F3: Induction 08:00

Day 1 (Feb 18 Wed)

   <Ann>                2/18/2009 (03:50)  Fermentation 19:10 
                             F1: OD (5.74) Contamination? (N)  
                             F3: OD (6.65) Contamination? (N)  F3: Induction 16:00 
   <Sung-Hye>           2/18/2009 (07:30)  Fermentation 22:50 
                             F1: Base consumption (400 ml - 340 ml = 60 ml)
   <Ann>                2/18/2009 (08:50)   Fermentation 24:00 
                             F3: OD (6.70) Contamination? (N) Complete Fermentation  F3: Induction 21:00 
                             F1: OD (7.20) Contamination? (N) F1: Induction 00:00 
                             Start feeding: (1 rpm)
File:Watsonpump.tif
   <Ann>                2/18/2009 (12:50)   Fermentation 25:00 
                             F1: OD (6.58) Contamination? (N)  F1: Induction 04:00 
   <Ann>                2/18/2009 (16:50)   Fermentation 29:00 
                             F1: OD (8.12) Contamination? (N)  F1: Induction 08:00 

Day 2 (Feb 19 Thu)

   <Ann>                2/19/2009 (00:50)  Fermentation 37:00 
                             F1: OD (8.64) Contamination? (N)  F1: Induction 16:00 
   <Ann>                2/19/2009 (06:50)  Fermentation 43:00 
                             F1: OD (9.04) Contamination? (N)  F1: Induction 22:00 
   <Sung-Hye>           2/19/2009 (09:30)  Fermentation 45:00 
                             Measure Base (400 ml - 270 ml= 130 ml total)
   <Ann>                2/19/2009 (13:30)  Fermentation 49:40 
                             F1: OD (10.36) Contamination? (N)  F1: Induction 28:40 
   <Ann>                2/19/2009 (17:50)  Fermentation 53:00 
                             F1: OD (10.00) Contamination? (N)  F1: Induction 33:00 
   <Ann>                2/19/2009 (23:50)  Fermentation 58:00 
                             F1: OD (10.72) Contamination? (N)  F1: Induction 38:00 

Day 3 (Feb 20 Fri)

   <Ann>                2/20/2009 (07:20)  Fermentation 65:30 
                             F1: OD (11.75) Contamination? (N)  F1: Induction 45:30 
   <Sung-Hye>           2/20/2009 (10:00)  Fermentation 68:10 
                             F1: Base (400 ml - 170 ml = 230 ml total)  F1: Induction 48:10 
   <Ann>                2/20/2009 (13:50)  Fermentation 72:10 
                             F1: OD (10.95) Contamination? (N)  F1: Induction 52:00 
   <Ann and Sung-Hye>   2/20/2009 (15:00)  Fermentation 73:10  
                             F1: Harvest F1: Induction 52:10

References

  1. www.ravenbiotech.com/product.html [Paper1]


  1. High Cell Density Fermentation of Saccharomyces cerevisiae JUL3 in Fed-batch Culture for the Production of β-Glucan (2007) 13, 1, 153-158 [Paper2]
  File:IE13-1-0153.pdf
  Fermentation condition: 30°C, 200 rpm, no pH control, 1 vvm
  
  1. Optimization of enterokinase fermentation using a recombinant Saccharomyces cerevisiae. (2005) 40, 717-722 [Paper3]
  File:PB-40-717.pdf
  Fermentation condition: 30°C, 500 rpm, no pH control, 2 vvm
  1. Alakomi HL, Paananen A, Suihko ML, Helander IM, and Saarela M. Weakening effect of cell permeabilizers on gram-negative bacteria causing biodeterioration. Appl Environ Microbiol. 2006 Jul;72(7):4695-703. DOI:10.1128/AEM.00142-06 | PubMed ID:16820461 | HubMed [Paper4]
  1. Peamiability test: Aliquots (100 µl) of this cell suspension were pipetted into fluoroplate wells, 
     which contained NPN (10 µM) and, as test substances, either EDTA (1.0 and 0.1 mM), PEI (10 µg ml–1), 
     DMSA (1 mM), AOT (1 mM), or HEPES buffer (control) to make up a total volume of 200 µl. If desired, 
     MgCl2 was added to the cell suspension before addition of NPN. Fluorescence was monitored within 
     3 min from four parallel wells per sample (excitation, 355 nm; half bandwidth, 38 ± 3 nm; emission, 
     402 nm; half bandwidth, 50 ± 5 nm). Each assay was performed at least three times. 
  1. Hounsa CG, Brandt EV, Thevelein J, Hohmann S, and Prior BA. Role of trehalose in survival of Saccharomyces cerevisiae under osmotic stress. Microbiology. 1998 Mar;144 ( Pt 3):671-80. DOI:10.1099/00221287-144-3-671 | PubMed ID:9534237 | HubMed [Paper5]
  1. OD600 unit is equivalent to 0.64 +/- 0.02 g cells (dry weight)/L
  1. Hao Q, Hong SH, and Maret W. Lipid raft-dependent endocytosis of metallothionein in HepG2 cells. J Cell Physiol. 2007 Feb;210(2):428-35. DOI:10.1002/jcp.20874 | PubMed ID:17111383 | HubMed [Paper6]
  1. To demonstrate the localization of the proteins relative to the cells, cell contours were visualized 
     with Alexa 647-concanavalin A.
  1. Drew D, Newstead S, Sonoda Y, Kim H, von Heijne G, and Iwata S. GFP-based optimization scheme for the overexpression and purification of eukaryotic membrane proteins in Saccharomyces cerevisiae. Nat Protoc. 2008;3(5):784-98. DOI:10.1038/nprot.2008.44 | PubMed ID:18451787 | HubMed [Paper7]
  1. Newstead S, Kim H, von Heijne G, Iwata S, and Drew D. High-throughput fluorescent-based optimization of eukaryotic membrane protein overexpression and purification in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 2007 Aug 28;104(35):13936-41. DOI:10.1073/pnas.0704546104 | PubMed ID:17709746 | HubMed [Paper8]
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