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Three aliquots of 200 μl from each flask were transferred to a sterile Costar 96-well black plate with flat clear bottom (Corning).
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Subsequently, the flask was transferred into a water bath set at a certain temperature.
After 24 h of growth, the entire broth from the growth flask was transferred to a conversion flask.
The desired amount of AuNPs-PSNPs was added and reaction flask was transferred to ultrasonic bath cleaner for 6 h.
An aliquot (100 μL) of the content of each Erlenmeyer flask was transferred to a microtube filled with 900 μL of sterile NaCl aqueous solution.
The flask was transferred to a preheated oil bath (270 °C) and heated for 6 h with mechanical stirring under nitrogen atmosphere.
Seed culture (300 ml), cultivated at 30°C for 17 h in shake flask was transferred to the fermenter containing 2.7 L of the fermentation medium to initial production in bioreactor.
The content of the flask was transferred to 30 mL Falcon tube and centrifuged at 4,400 rpm for 10 min.
When the cells were grown to early stationary phase, 5 mL of culture broth from the preceding flask was transferred to a new flask culture with the same medium composition.
Allowing the growth to reach to the early stationary phase, 5 ml of culture broth from the preceding flask was transferred to a new flask culture with the appropriate medium composition.
The mixture was heated in a microwave (280 W) for 5 min. Thereafter, the flask was transferred to the ultrasonic bath (Branson Ultrasonic, Danbury, USA) and the frequency and power was adjusted to 20 kHz and 200 W, respectively.
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