Experimental
        Arthrobacter viscosus NRRL B-1973 was obtained from the American Type Culture Collection (ATCC 19584), Rockville, MD.
 

Bacterial Growth Conditions - Modified Haggstrom Liquid Medium
Item
g·l-1
peptone
1.0
yeast extract
1.0
Na2HPO4
0.6
KH2PO4
0.4
MgSO4.7H2O
0.2
NH4Cl
0.1
 
Trace Elements
micromole/l
FeSO4.7H2O 0.036
H3BO3 0.097
CoCl2.6H2O 0.017
Cu2SO4.5H2O 0.08
MnSO4.H2O 0.019
ZnSO4.7H2O 0.008
 
1. Adjusted to pH 8.0 with 10N KOH 3. Cultured in 2 or 4 L flasks @ 28 °C
2. glucose 2% w/v 4. Using an orbital shaker @200 rpm
 
 

Enzyme Purification Procedure
1.  Harvested in early stationary phaseby centrifugation @  15,000 x g , 20min 4. Lysed using lysozyme (Sigma) 0.1 mg /ml, 4°C, 2 hr;  rapid freeze thaw cycle, add  MgCl2, 
2.  Wash in Tris-HCl  (50 mm pH 7.2) 5.  Then DNase I (25 micromol/ml, 4°C, 30 min), and centrifuge 25,000 x g @  4°C to remove cellular debris
 3.  Resuspend in lysis buffer: Tris-HCl (50mm, pH 7.2), EDTA (10 mM, Triton X-100 (0.1% v/v), 0.5 g cells/ml