James Hood, 70, died in his hometown of Gadsden, Alabama, according to his obituary notice published on Friday.
Hood and Vivian Malone were the first African-American students to register to attend the all-white state university, and did so over the objections of a governor known for his resistance to the U.S. civil rights movement.
Fifty years ago this week, on January 14, 1963, Wallace promised "segregation now, segregation tomorrow, segregation forever" during his inaugural speech at the Capitol in Montgomery.
Five months later, on June 11, 1963, Wallace stood outside an auditorium at the University of Alabama in Tuscaloosa in a failed effort to prevent Hood and Malone from registering.
The governor's attempt was thwarted by President John F. Kennedy, who ordered Alabama National Guard units to the campus. Wallace then stepped aside, allowing the students to enter.
Hood "was willing to put his life on the line that day, but he wasn't afraid. He wanted to create opportunities for others," said Samory Pruitt, vice president for community affairs at the University of Alabama.
Hood attended the university for a few months before moving to Michigan and going to school there. He returned in 1997 to earn his doctorate in education, according to the university.
In 2010, the university honored his pioneering efforts by naming the site of the infamous confrontation the Malone-Hood Plaza.
Hood met with Wallace in 1996, and the former governor apologized for his actions. When Wallace died in 1998, Hood attended the funeral, calling the former segregationist a changed man.
(Reporting by Verna Gates; Editing by Colleen Jenkins, Cynthia Johnston and Cynthia Osterman)