Table 1 Comparison of different cell disintegration techniques in the recovery of GAD

Enzymatic lysis

Yatalase^® (3 mg/ml, 30% w/v cell concentration, 30 min, 60˚C)

0.4 ± 0.01

0.61 ± 0.02

0.70 ± 0.02

Chemical permeabilization

Solvent permeabilization

Toluene, 10% (v/v), 120 min, 25°C

0.09 ± 0.02

0.14 ± 0.03

0.97 ± 0.05

Chloroform, 10% (v/v), 120 min, 25°C

0.15 ± 0.04

0.17 ± 0.07

0.88 ± 0.06

Butanol, 10% (v/v), 60 min, 25°C

0.06 ± 0.02

0.08 ± 0.04

0.78 ± 0.02

Ethanol, 10% (v/v), 120 min, 25°C

0.08 ± 0.01

0.19 ± 0.05

0.42 ± 0.04

Acetone, 10% (v/v), 60 min, 25°C

0.04 ± 0.03

0.17 ± 0.04

0.24 ± 0.07

Detergent permeabilization

Triton X-100, 0.2% (v/v), 120 min, 25°C

0.13 ± 0.04

0.14 ± 0.07

0.93 ± 0.08

CHAPSO, 0.2% (v/v), 120 min, 25°C

0.10 ± 0.07

0.17 ± 0.02

0.62 ± 0.04

Tween 80, 0.2% (v/v), 120 min, 25°C

0.09 ± 0.02

0.26 ± 0.05

0.36 ± 0.05

SDS, 0.2% (v/v), 120 min, 25°C

0.04 ± 0.02

0.15 ± 0.01

0.28 ± 0.08

Mechanical disruption

Sonication (10% w/v cell concentration, 50% pulser, 30 W, 10 min)

6.7 ± 0.02

3.40 ± 0.04

1.99 ± 0.38

High speed Homogenization (10% w/v cell concentration, 18,000 rpm, 30 min)

3.52 ± 0.02

14.0 ± 0.01

0.25 ± 0.03