2.2. Culture systems
The nutrient spray bioreactor (NSB): The NSB system has developed by us consists of two vessels, each constructed from 1 L glass
reagent bottles – an upper culture chamber, and a lower medium
reservoir (Fig. 1a). The culture chamber is 12 cm in height with
9.5 cm inner diameter, and has 2.5 cm wide rims at the top and
bottom. It has a lid (diameter: 14.5 cm) with a brass spray nozzle
(diameter: 0.8 cm, length: 3.7 cm, spray orifice: 0.5 mm) attached.
The medium reservoir is 10 cm in height and 9.5 cm in diameter,
and has a 2.5 cm wide rim at the top, an air inlet, and a medium
outlet, which is connected to the spray nozzle by a silicone tube
(ID 0.6 mm). Nutrient spray was created by releasing compressed
air into the medium reservoir via a hydrophobic membrane filter
(0.22m) connected to the air inlet. There is a one-way check valve
at the bottom of the culture chamber, which allows the sprayed liquid to return to the medium reservoir but prevents reverse flow of
liquid or air from the medium reservoir to the culture chamber.
The semi-solid and liquid culture systems: For the semi-solid
culture system (Fig. 1b) the protocol described inPiao et al. (2003)
was followed. For the liquid culture system (Fig. 1c) a combined
support of stainless steel net and filter paper (Whatman No. 1) was
used and for sterile air exchange the caps of the culture vessels
were fitted with 0.22m hydrophobic filters
2.2. Culture systemsThe nutrient spray bioreactor (NSB): The NSB system has developed by us consists of two vessels, each constructed from 1 L glassreagent bottles – an upper culture chamber, and a lower mediumreservoir (Fig. 1a). The culture chamber is 12 cm in height with9.5 cm inner diameter, and has 2.5 cm wide rims at the top andbottom. It has a lid (diameter: 14.5 cm) with a brass spray nozzle(diameter: 0.8 cm, length: 3.7 cm, spray orifice: 0.5 mm) attached.The medium reservoir is 10 cm in height and 9.5 cm in diameter,and has a 2.5 cm wide rim at the top, an air inlet, and a mediumoutlet, which is connected to the spray nozzle by a silicone tube(ID 0.6 mm). Nutrient spray was created by releasing compressedair into the medium reservoir via a hydrophobic membrane filter(0.22m) connected to the air inlet. There is a one-way check valveat the bottom of the culture chamber, which allows the sprayed liquid to return to the medium reservoir but prevents reverse flow ofliquid or air from the medium reservoir to the culture chamber.The semi-solid and liquid culture systems: For the semi-solidculture system (Fig. 1b) the protocol described inPiao et al. (2003)was followed. For the liquid culture system (Fig. 1c) a combinedsupport of stainless steel net and filter paper (Whatman No. 1) wasused and for sterile air exchange the caps of the culture vesselswere fitted with 0.22m hydrophobic filters
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