3. A newly appointed biochemical engineer was tasked with inoculum preparation and scale up of a culture of a sensitive

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3. A newly appointed biochemical engineer was tasked with inoculum
preparation
and scale up of a culture of a sensitive bacterium strain. They
undertook the
following operating procedure:
Step 1: They prepared the working culture of the bacterium on an
agar slant,
and waited for 1 day
Step 2: They added saline and glass beads to the slant, and waited
for 1 day
Step 3: They transferred the culture to a shake flask preloaded
with fresh agar,
and waited for 1 day
Step 4: They transferred the culture to a seed fermenter and waited
for 1 day
Step 5: They transferred the fermenter contents to the production
fermenter
(a) What was the purpose of the 1 day waiting time between steps?
Use an
appropriate sketch to support your explanation.

(b) Tests carried out on the production fermenter indicated that
the cell mass
concentration was far below the level expected. Review the
engineer’s
operating procedure and identify three possible reasons for
this.

(c) Consider the relevance of the five pillars of GMP to the
scenario detailed in
this question and propose one specific improvement for each
pillar.

(d) Out of the improvements you proposed in part (c), which do you
think is the
most important? Justify your choice.