Commander Barry "Butch" Wilmore and flight engineer Terry Virts began their six-and-a-half hour spacewalk at 6:51 am (12:51 GMT) when they placed their suits on internal battery power before emerging from the airlock, NASA said.
The astronauts are both wearing helmet cameras that allow viewers on Earth to see the spacewalkers' perspective, as part of a live broadcast on NASA television and on its website.
The team's goal for the spacewalk is to route two bundles of cable, lubricate latching parts of the space station's robotic arm and prepare for another operation later this year to move modules around the orbiting outpost.
The whole robotic arm process will take about three hours, said NASA commentator Rob Navias.
Launched in 2001, the robotic arm is regularly used to move equipment and astronauts around the exterior of the space station.
It also reaches out to grab the arriving unmanned space capsules bringing supplies and science experiments to the space station.
Navias described the robotic arm as "the workhorse for the assembly of the space station," and said parts of it have "grown a bit arthritic."
Boeing plans to send its first astronaut to space aboard the CST-100 spaceship in late 2017, followed by SpaceX soon after.
During the first outing by Wilmore and Virts on Saturday, the pair routed 364 feet (110 meters) of cable, with goal of setting up 700 feet by the end of the final spacewalk of this series on Sunday.
Several more spacewalks in the coming months will help the station get ready for the arrival of a pair of international docking adapters that will be delivered later this year.
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