Cell disruption
Cellular disruption of P. cruentum was performed using a bead mill (Dyno Mill-
Multi Lab, Muttenz, Switzerland) equipped with a cooling jacket that allowed to
maintain constant the temperature at 14 ◦C through processing. The release of BPE
from P. cruentum using the bead mill device has not been reported in literature,
and therefore a characterization of the effect of different process parameters on BPE
release was necessary. Loaded cell suspension consisted on 50 mM dibasic phosphate
potassium buffer with 20% (w/w) P. cruentum cells. Cell suspension was fed to
the bead mill using a variable speed peristaltic pump (Manostat Vera Pump, Thermo
Scientific) at a constant flow rate of 6 cm3/s. The grinding chamber was initially filled
with a 70% (v/v) bead load.
In order to characterize and optimize P. cruentum disruption using a bead mill,
experiments were first done to determine the effect of rotational speed and bead
size in BPE liberation yield. Rotational speeds of 2300, 2900 and 4000 rpm were
employed. In order to determine the effect of bead size on BPE release efficiency
two different bead diameters (0.5 and 1.2 mm) were tested. Once optimal rotation
speed and bead size were determined, the effect of bead load (60–80%, v/v) and
biomass load (10–30%, w/w) were studied. The homogenate obtained from the cell
disruption stage (including cell debris) was referred to as BPE crude extract. BPE
release constant (k) under optimal process conditions was calculated using Eq. (1)
[21], where t is disruption time, R is the release of BPE at time t, and Rm is the
maximum achievable BPE release (1.5 mg BPE/g wet biomass, considering that after
25 min of cell disruption BPE release was constant in this value, hence being the
maximum observed yield).
Cell disruption
Cellular disruption of P. cruentum was performed using a bead mill (Dyno Mill-
Multi Lab, Muttenz, Switzerland) equipped with a cooling jacket that allowed to
maintain constant the temperature at 14 ◦C through processing. The release of BPE
from P. cruentum using the bead mill device has not been reported in literature,
and therefore a characterization of the effect of different process parameters on BPE
release was necessary. Loaded cell suspension consisted on 50 mM dibasic phosphate
potassium buffer with 20% (w/w) P. cruentum cells. Cell suspension was fed to
the bead mill using a variable speed peristaltic pump (Manostat Vera Pump, Thermo
Scientific) at a constant flow rate of 6 cm3/s. The grinding chamber was initially filled
with a 70% (v/v) bead load.
In order to characterize and optimize P. cruentum disruption using a bead mill,
experiments were first done to determine the effect of rotational speed and bead
size in BPE liberation yield. Rotational speeds of 2300, 2900 and 4000 rpm were
employed. In order to determine the effect of bead size on BPE release efficiency
two different bead diameters (0.5 and 1.2 mm) were tested. Once optimal rotation
speed and bead size were determined, the effect of bead load (60–80%, v/v) and
biomass load (10–30%, w/w) were studied. The homogenate obtained from the cell
disruption stage (including cell debris) was referred to as BPE crude extract. BPE
release constant (k) under optimal process conditions was calculated using Eq. (1)
[21], where t is disruption time, R is the release of BPE at time t, and Rm is the
maximum achievable BPE release (1.5 mg BPE/g wet biomass, considering that after
25 min of cell disruption BPE release was constant in this value, hence being the
maximum observed yield).
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