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Several mutants (1-4) are isolated, all of which require compound E for growth. The compounds A to D in the biosynthetic pathway to E are known,
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Growth will be supported by a particular compound if it is later in the pathway than the enzymatic step blocked in the mutant. Restated, the more mutants a compound supports, the later in the pathway growth must be. In this example, compound E supports growth of all mutants and can be considered the end product of the pathway. Alternatively, compound A does not support the growth of any mutant and can be considered the starting substrate for the pathway.