bioreactor studies of heterologous protein production by ...

1) Reagent solution preparation was carried out as foiiows. Enzyme Solution was
prepared by adding contents of I capsule of PGO enyme$ No. 5106, to 1ûûm.I
distïiled water in an amber bottle. Invert bottie several times with gentle shaking to
dissolve. Color Reagent Solution was prepared by recoastituting one vial of o-
Dianisidine Dihydrochloride, Catalogue No. 510-50, with 20 ml water. Combined
EnzymeColor Reagent Solution was prepared by combining 100 ml Enzyme Solution
and 1.6 ml Color Reagent Solution. The solutions were mixed by inverdng several
times or with rnild shaking. 1OOO m& Glucose Standard Solution was supplied ready
to use.
2) Label the testing tubes as foUows: BLANK, STANDARD, TEST 1, EST 2, etc.
3) To BLANK tube add 0.5 ml water. To STANDARD tube add 0.5 ml of a 20-fold
dilution of 1 g/L Glucose Standard Solution. To TEST tubes add 0.5 mi of a suitable
diluted samples (not greater than 100 mg fi).
4) To each tube add 5.0 ml of Combined EnzymeColor Reagent Solution and mix each
tube thoroughly.
5) Incubate aü tubes at 37 O C for 30 minutes in water bath.
6) At the end of incubation period, remove ail tubes h m water bath, nad absorbante
[A] of STANDARD and TESTS at 450 nm, using BLAMC as reference.
7) Caldate TEST values as foïlows:
Glucose [mg / L] = x50
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