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The Family Guy creator, who voices Peter and Stewie Griffin on the comedy, will appear at the Royal Albert Hall in London on August 27.
MacFarlane is taking part in a show paying homage to Broadway musicals past and present.
The 38-year-old, who also pens American Dad, will perform with the John Wilson Orchestra.
MacFarlane will release his debut album, a collection of big band arrangements, later this year.
The star is currently remarking vintage cartoon The Flintstones for Fox.
Watch Seth MacFarlane sing 'Two Sleepy People' with Sara Bareilles below:
