ONE GENE-ONE ENZYME HYPOTHESIS

BEADLE AND TAUTUM EXPERIMENT

✓ The one gene, one enzyme hypothesis is the idea that each gene encodes a single enzyme. Though, this
idea is generally not correct.
✓ Sir Archibald Garrod, a British medical doctor, was the first to suggest that genes were connected to
enzymes.
✓ Beadle and Tatum confirmed Garrod's hypothesis using genetic and biochemical studies of the bread
mold Neurospora.
✓ Beadle and Tatum identified bread mold mutants that were unable to make specific amino acids. In
each one, a mutation had "broken" an enzyme needed to build a certain amino acid.
✓ Typical gene provides instructions for building a protein, which in turn determines the observable
features of an organism.
✓ For instance, Gregor Mendel's flower color gene specifies a protein that helps make pigment molecules,
giving flowers a purple color when it works correctly.
✓ Mendel, however, did not know that genes (which he called "heritable factors") specified proteins and
other functional molecules. In fact, he didn't even speculate about how genes affected the observable
features of living organisms.

Beadle and Tatum: Connecting genes to enzymes

✓ George Beadle and Edward Tatum, carried out a series of experiments in the 1940s.
✓ Beadle and Tatum worked with a simple organism: common bread mold, or Neurospora crassa.
✓ Using Neurospora, they were able to show a clear connection between genes and metabolic enzymes.

One Gene-One Enzyme Hypothesis-

✓ Beadle and Tatum made it clear that genes are the functional units and transmitted to progenies over
generations; also, they undergo mutations.
✓ Wild-type Neurospora crassa (red bread mold) is prototrophic.
✓ Prototrophs are able to grow on minimal medium.
✓ Minimal medium contains only inorganic salts, simple carbon source, and water.
✓ Auxotrophs require nutritional supplement(s) for growth.
✓ N. crassa treated with X-rays and selected for X-ray induced mutations that would have been lethal.
✓ Conidia are mutagenized with X-ray.
✓ Their selection would have been possible when N. crassa was allowed to grow on nutrient medium
containing vitamin B6.
✓ Possible mutants are crossed to wild-type and individual ascospores are cultured on complete
(permissive) medium.
✓ Cultures are plated on minimal medium and auxotrophs are isolated.
✓ Auxotrophs are tested for nutritional requirement.
✓ Each requirement for a specific nutrient (mutant effect) followed Mendelian pattern of inheritance.
✓ Beadle and Tatum deduced that each mutant was deficient for an essential enzyme in a biosynthetic
pathway.
✓ This explains that X-rays mutated vitamin B6 synthesizing genes.
✓ They concluded that a gene codes for the synthesis of one enzyme.
✓ In 1958, Beadle and Tatum with Lederberg received a Nobel prize for their contribution to
physiological genetics.
 Figure 1: Two or more are enzymes are involved in formation of a product

Figure 2: Diagrammatic Representation of Beadle and Tatum Experiment

Result of the Beadle and Tautum Experiment-

✓ The cells could grow on minimal medium, a nutrient source with just sugar, salts, and one vitamin
(biotin).
✓ Neurospora cells can survive on this medium, while many other organisms cannot.
✓ That's because Neurospora has biochemical pathways that turn sugar, salts, and biotin into all the
other building blocks needed by cells (such as amino acids and vitamins).
✓ Neurospora cells grow well on complete medium, which contains a full set of amino acids and
vitamins.
✓ After inducing X-rays, a few colonies grew normally on complete medium, but couldn't grow at all
on minimal medium.
✓ These were the nutritional mutants that Beadle and Tatum had predicted.
✓ On minimal medium, each mutant would die because it could not make an particular essential
molecule out of the minimal nutrients.
✓ Complete medium would "rescue" the mutant (allow it to live) by providing the missing molecule,
along with a variety of others.
 ✓ First, they grew each mutant on minimal medium supplemented with either the full set of amino
acids or the full set of vitamins (or sugars, though we won't examine that case here).
✓ If a mutant grew on minimal medium with amino acids (but not vitamins), it must be unable to make
one or more amino acids.
✓ If a mutant grew on the vitamin medium but not the amino acid medium, it must be unable to make
one or more vitamins.
✓ Start with a nutritional mutant. By definition, the nutritional mutant can grow on complete medium,
but not on minimal medium.

Conclusion-
✓ In this way, Beadle and Tatum linked many nutritional mutants to specific amino acid and vitamin
biosynthetic pathways.
✓ Their work produced a revolution in the study of genetics and showed that individual genes were
indeed connected to specific enzymes.