Aluko previously played for Birmingham City, Charlton Athletic, and Chelsea in England's FA Women's Premier League.

She played for Saint Louis Athletica, Atlanta Beat, and Sky Blue FC in the American Women's Professional Soccer (WPS) from 2009 to 2011.

[4] Born in Lagos, Nigeria, to Gbenga and Sileola, Aluko moved with her family to Birmingham in the West Midlands region of England at the age of six months.

[19] She helped Charlton defeat Fulham 1–0 to win the FA Women's Premier League Cup at Underhill Stadium in March 2004.

[20][21] During the 2003–04 season, Aluko appeared as a second-half substitute when Charlton lost the FA Women's Cup final to Arsenal, 3–0 at Loftus Road in May.

[25] Pace was considered one of her attributes and this was evident when she sprinted onto a through ball from Emma Coss to score the winning goal in the 2005 FA Women's Cup Final at Upton Park.

[27] Aluko scored two goals in the first half of Charlton's 2–1 victory over Arsenal in the 2006 FA Women's Premier League Cup Final at Adams Park.

[37] In 2012, Aluko registered five goals and two assists in 17 FA WSL League and Cup appearances, as Birmingham City finished runners up to Arsenal in both competitions.

[40] Birmingham did defeat Chelsea 3–2 on penalties, after it was 2–2 at the end of extra time, in the 2012 FA Women's Cup Final on 26 May 2012 at Ashton Gate to win the club's first major honour in its 44-year history.

[44][45] Having had a poor season domestically,[15] Chelsea reached the final of the invitational 2013 International Women's Club Championship, but lost 4–2 to INAC Kobe Leonessa.

[55][56] In 2015, Aluko played in the first FA Women's Cup Final held at Wembley Stadium on 1 August 2015 in front of a record attendance of 30,710.

Individually, Aluko was at the heart of Chelsea's win as she put in a Player-of-the-Match performance and provided the assist for Ji So-yun's 39th-minute winner.

[62] However, in 2019 she spoke about negative experiences in the city, calling it "decades behind" and saying she had been made to feel like Pablo Escobar on occasions at Turin airport.

"[65] She returned to Britain in December having won Serie A, Coppa Italia and Supercoppa Italiana, as well as being Juventus' top scorer for last season.

[75] In the final group game against Sweden Aluko almost scored a bizarre equaliser, but was left disappointed as hosts England lost 1–0 and exited the competition.

[76] At the FIFA Women's World Cup 2007, Aluko featured in group matches against Japan and Argentina, as well as the 3–0 quarter-final defeat by the United States.

[77] After the tournament, she was critical of The Football Association and the level of financial support provided to England's top female players.

[78] Aluko featured much more prominently at UEFA Women's Euro 2009, scoring in the group match win over Russia[79] and adding two more in the quarter-final victory over hosts Finland.

[83] During the final tournament, Aluko responded angrily to public criticism of her performance in the 1–1 draw with Mexico, in which she wasted multiple goalscoring opportunities.

The Football Association later publicly apologised to Aluko at the select committee hearing of the Department of Culture, Media and Sport (with some DCMS MPs calling for relevant FA officials to resign over their handling of the case)[94] after the same barrister found racist remarks were made to her and teammate Drew Spence, following a third investigation.

When England's entire team—led by black forward Nikita Parris—pointedly ran to celebrate with coach Sampson after scoring in their next match against Russia, Aluko publicly criticised her former team-mates, accusing them of selfishness, lacking respect and requiring diversity training.

[97] Parris later apologised to Aluko in an open letter in June 2020 admitting that the celebration with coach Sampson was a "thoughtless action" that showed a lack of empathy, understanding and ignorance given that Sampson was under investigation for making racially discriminatory remarks to Aluko and fellow player Drew Spence of which he was found guilty and for which the FA later apologised.

[108] She made her Great Britain debut, in its first ever official fixture, in a friendly goalless draw with Sweden at the Riverside Stadium, Middlesbrough on 20 July 2012, starting and playing the entire match.

Aluko also started as Britain lost 2–0 to Canada in the quarter-finals, courtesy of two early goals, and ended hopes of at least reaching a match for an opportunity at a medal.

[122] In May 2021, Aluko became the sporting director of Los Angeles-based Angel City FC ahead of their inaugural season in the American National Women's Soccer League (NWSL).

[126] In July 2024, Eni Aluko launched a £60,000 scholarship in conjunction with Brunel University, supporting the next generation of lawyers and sports stars in their studies and career development.

[130] Her younger brother Sone is a retired footballer who played most recently for Ipswich Town and was an England youth international, but in May 2009 accepted a call-up to represent Nigeria.

[142] Aluko was appointed Member of the Order of the British Empire (MBE) in the 2023 Birthday Honours for services to association football and charity.