A =

Ec I The reaction volume = 1 .

25mL The volume of enzyme
=

55M

= S A =

& Sc I

The information that we will use :

initial of change (A) 0 85 mint

rate of
=

we use the Absorbance

.

+ +
& =
6220M cm

The question wants us to :

0 calculate the rate of NADH Mmint Beer Lambert

in using
⑦ calculate triAl mal of NADH
appearing per minute from total volume of reaction mixture
-
> Invest into mmulment

⑦
anvest mmalment to cone unit Amount of enzyme that Immot substrate minute)
Units
catalyses
=

per

① SA =

E1C L

+ 0 . 85 mint =

(6220M/C) (AC) ( 1 CM)

a
5 min
1 c
=
1 . 367 x 10 M mint

② In this step we want to convert the M part to mot , then umol

: we use reaction mixture volume

1 c =
1 367 x 15-4 Mmin
G
.

This is the same as :

1 2 =
1 367
.
x 10 mot L" mint

Lt
Get rid of by multiplying with the reaction mixture volume

Renation mix. . Volume

=
1 .
20 mL
=
1 2.
x 10-3 L

=> 1 .

367 x 10-4 mo mint x 1 2

.
x 10-3
-
1 .
64 x 10-7 mol mint

C
3 convert to umalmint CI know instructions
say
mmul , but its easier to concert to umol)
&

S
1 64 x 10-7 mot min-

!
.

(1 mol ·
1000000 und)

. = .
0 164 umolmin-
(this is
why)
④ Now
, we need convert
to into UNITS =

Cone unit =
Amount of enzyme that catalyses Immot substrate per minute)

divide of umalmint
we well the rele
by volume of enzyme in me to

unity
get
:

: umamin
=
2 . 981 UnitemL" FINAL ANSWER
0 . 055 mL