The lac operon

2.1.5

Enzyme induction
Enzymes involved in basic cellular functions are synthesised at a fairly constant rate. Inducible enzymes are synthesised at varying rates, according to the cells circumstances. Bacteria adapt to their environments by producing enzymes to metabolise certain nutrients only when those nutrients are present. E.coli normally respires glucose but it can also use lactose as a respiratory substrate. E.coli grown in a culure medium with no lactose can be placed in a medium with lactose. At first they cannot metabolise the lactose because they only have tiny amounts of the two proteins needed to metabolise it. B-galactosidase catalyses the hydrolysis of lactose to glucose and galactose, and the protein lactose permease transports lactose into the cell. Once lactose is added the bacteria can increase the synthesis of the two proteins by about 1000 times. Lactose must trigger the production of the two proteins, and is known as the inducer. However, production is only triggered if glucose is absent.

Lac genes form an operon

The lac operon is a section of DNA within the bacteriums DNA. It consists of a number of parts: The structural genes: Z codes fo the enzyme b-galactosidase, and Y codes for lacose permease . Each consists of a sequence of base pairs that cn be transcribed into a length of mRNA. The operator region: O, is a length of DNA next to the structural genes. It can switch them on or off. The promoter region: P, is a length of DNA to which the enzyme RNA polymerase binds to begin the transcription of the structural genes, Z and Y. The regulator gene, I, is not part of the operon and is some distance from the lac operon.

How it works
When lactose is absent from the growth medium. The regulator gene is expressed and the repressor protein is synthesised. It has two binding sites, one that binds to the lactose and one that binds to the operator region. The repressor protein binds to the operator region. In doing so it covers part of the promoter region, where RNA polymerase normally attatches . RNA polymerase cannot bind to the promoter region so the structural genes cnnot be transcribed into mRNA. Without mRNA these genes cannot be translated and bgalactosidase and lactose permease cannot be synthesised.

Continued...
When lactose is added to the growth medium Lactose molecules bind to the other site on the repressor protein. This causes the molecules or repressor protein to change shape so that it's other binding site cannot now bind to the operator region. The repressor breaks away from the operator region. This leaves the promoter region unblocked. RNA polymerase can now bind to it ad initiate the transcription of mRNA for genes Z and Y. The operator-repressor-inducer system acts as a molecular switch. It allows transcription and subsequent tranlation of the structural genes, Z and Y, into the lac proteins b-galactosidase and lactose permease As a result E.coli bacteria can use lactose permease to take up lactose from the medium, into their cells. They can then convert the lactose to glucose and glactose using the b-gaactosidase enzyme. These sugars can then be used for respiration, thus gaining energy from lactose.

Questions
Structural genes - codes for lac proteins Regulator gene - produces repressor protein Promoter - binds to RNA polymerase Operator - binds to repressor What is the function of the repressor protein? It binds to the operator region and blocks the promoter region so RNA polymerase can't bind and therefore genes Z and Y can't be transcribed. What is the role of lactose? It binds to repressor at another site. It causes it's shape to change so it moves away from operator and leaves the promoter region free for RNA polymerase to bind, Z and Y genes are transcribed. Explain the advantage of having a lac operon to induce enzymes It only makes enzymes to metabolise lactose when lactose is present in the culture medium environment. The enzymes are inducible so resources aren't wasted.