The 31-year-old former Free State Stars and Mamelodi Sundowns defender joins the club on a permanent transfer having previously spent three seasons on loan for Babina Noko while contracted to the Brazilians.
He has also played for Stellenbosch and Chippa United on loan but signs for Sekhukhune after a season with Siwelele.
In his first spell with Sekhukhune, Mobbie made 90 appearances, scoring one goal and providing two assists.
A club statement confirmed the news: "Sekhukhune United Football Club is delighted to welcome Nyiko Mobbie back to Babina Noko on a permanent transfer.
"Mobbie returns to the club after a highly successful spell between the 2021/22 and 2023/24 seasons, during which he established himself as one of the Premier Soccer League's standout performers.
"His impressive displays earned him multiple call-ups to the South African senior national team and was part of the team that earned third-place finish at the 2023 Africa Cup of Nations.
"A dynamic and experienced defender, Mobbie returns with valuable top-flight and international experience, further strengthening the squad ahead of the 2026/27 Betway Premiership campaign.
"Everyone at Sekhukhune United warmly welcomes Nyiko back to Babina Noko and wishes him every success in the club's colours once again."