Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0478
trehalose synthase

Lot Information:

Purification Lot: PC010478-41-02-05
sPlasmid ID: PC010478-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Inoue Y.
Purified Date: 10/19/2005
Total Protein Yield: 1.8 mg
Final Volume: 0.14 mL
Conc.: 12.9 mg/mL


Culture Lot:


host cell



07/05/2005B834(DE3)6 L

Cell Lysis and Heat Treatment:

24.0g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 500mM NaCl (pH8.0)
Heat Treatment at 70°C for 10 min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)6 
Super Q Toyopearl 650M 30 ml20 mM Tris-HCl (pH 8.0)40.22M NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)40.29M NaCl
Hydroxyapatite CHT5-I10 mM KPi (pH 7.0)2flowthrough
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl0.551.3ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10