Calgary's Allison Beveridge, Vancouver's Laura Brown and Jasmin Glaesser and Kirsti Lay, of Medicine Hat, Alta., earned Canada its third gold in the sport with a time of 4:19.664 in the head-to-head final to defend their 2011 Pan Am title and reset their own Pan Am record.
On Thursday, the Canadians had established a new record in qualifications, which they improved on by nearly five seconds in the final race.
The American team, which includes Jennifer Valente, Sarah Hammer, Kelly Catlin and Lauren Tamayo, were no match for the host nation, clocking in well behind at 4:26.426.
Canada's women's pursuit team has had an impressive streak of success following their bronze- medal victory at the London Olympics in 2012, reaching the podium in every major event they've entered since.
Mexico earned bronze by beating Colombia.