The teams, who have vastly different international experience, were paired when the CAF Confederation Cup play-offs draw was made in Cairo on Tuesday.
Based in the southern Congolese city of Lubumbashi, Mazembe have been competing in Africa since 1967, winning 11 titles, including the marquee CAF Champions League five times.
Royal were promoted to the South African top flight only last year and are making their first appearance in a continental competition.
Without a CAF title since 2016, Mazembe started this season in the Champions League, but were relegated to the Confederation Cup after a shock loss to Ugandan side Vipers last weekend.
The Congolese reacted by firing French coach Franck Dumas and putting former Mazembe star Pamphile Mihayo in charge a third time.
Mihayo aims to win the Confederation Cup a third time after successes in 2016 and 2017 before Moroccan clubs became dominant, winning four finals and finishing runners-up in the other.
Rookies Royal upset Zesco United of Zambia in the second Confederation Cup qualifying round after a routine victory over Mbabane Highlanders of Eswatini.
All three Royal goals in Africa came from veterans with Ruzaigh Gamildien, 33, scoring twice and Mozambican Elias 'Domingues' Pelembe, 38, once.
Nigerians to face Libyans
To score against Mazembe, who lost to Inter Milan in the 2010 Club World Cup final, they must pierce a defence likely to include DR Congo international Issama Mpeko, 33.
The other South African hopefuls were paired with north African clubs -- Marumo Gallants meet Al Ahly Tripoli of Libya while Cape Town City face USM Alger of Algeria.
Nigerian contenders Rivers United and Plateau United, both Champions League losers in the final qualifying round, drew Libyan opponents.
Rivers play Al Nasr and Plateau come up against Al Akhdar in the first legs on November 2, with the return matches seven days later.
Title-holders Renaissance Berkane of Morocco continue a quest for a record-equalling third Confederation Cup title against US Monastir of Tunisia.
Draw
Rail Kadiogo (BUR) or V Club (COD) v St Eloi Lupopo (COD)
Royal Leopard (ESW) v Real Bamako (MLI)
TP Mazembe (COD) v Royal AM (RSA)
Primeiro Agosto (ANG) v Future (EGY)
ASEC Mimosas (CIV) v SC Gagnoa (CIV)
Djoliba (MLI) v FAR Rabat (MAR)
Al Ahly Tripoli (LBA) v Marumo Gallants (RSA)
ASKO Kara (TOG) v CS Sfaxien (TUN)
Young Africans (TAN) v Club Africain (TUN)
Flambeau Centre (BDI) v DC Motema Pembe (COD)
Rivers Utd (NGR) v Al Nasr (LBA)
US Monastir (TUN) v Renaissance Berkane (MAR, holders)
Cape Town City (RSA) v USM Alger (ALG)
Nigelec (NIG) v Pyramids (EGY)
La Passe (SEY) v Diables Noirs (CGO)
Plateau Utd (NGR) v Al Akhdar (LBA)
First legs: Nov 2, second legs: Nov 9
Note: The 16 winners qualify for group stage from Feb 12
