Global Christian campaigners, CitizenGo launched the online petition on 22nd of October, calling for the Reading branch to remain open after the decision was made not to renew the restaurant's lease following a number of demonstrations on site by LGBT activists.
Members of the LGBT community have taken issue with the organisation following comments made by the founder's Christian son.
Both chief executive Dan Cathy and his late father Samuel, who founded the chain in 1946, have publicly made donations towards Christian charities that oppose LGBT lifestyles.
In 2012, Dan Cathy confirmed that the company does not support same-sex marriage because it opposes the organisation's Christian values.
The petition claims that the "forced" closure of the restaurant is "an attack on religious freedom."
"Chick-Fil-A are not in breach of any UK employment laws and do not discriminate against any of their customers therefore there is no good reason why this popular restaurant cannot be allowed to remain open," it reads.
"It is not the role of a commercial landlord to police the corporate values or religious beliefs of their tenants. No company is compelled to engage in any type of philanthropic activity and it is entirely up to individuals and corporations to decide which organisations best reflect their values."
At the time of writing the petition has 17,300 signatures.
A second branch of the popular fast-food chain opened earlier this month in Scotland to similar opposition from the LGBT community.
The Edinburgh branch has prompted an MSP boycott of the Macdonald Aviemore Resort hotel and a petition calling for Macdonald Hotels not to support the firm.
LGBT rights campaigner, Scott Cuthbertson who launched the petition says Chick-Fil-A have "given millions to organisations that have opposed marriage and other rights for LGBT people, and calls on MacDonald's "not to support people who fund groups opposed to LGBT equality."
The petition has to date been signed by 825 people.
A spokeswoman for Macdonald Aviemore Resort told The Press and Journal: "Chick-fil-A is an enormously successful business, with over 2,300 restaurants in the US, and we are pleased to have them invest in the Aviemore economy, where the restaurant is proving extremely popular.
"It's vital to underline that, in both our recruitment and our customer care, we treat everyone with respect, regardless of race, religion, sexuality or gender."
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