The petition, which has already received over 1,000 signatures, is demanding the chain close its first ever Scottish eatery, which quietly opened inside the Macdonalds Hotel Resort at Aviemore last month.
The opening follows the US food chain's unsuccessful attempt to open its first UK outlet in Reading, which is set to close after its licence was revoked by The Oracle shopping centre following protests over the company's consistent funding of anti-LGBTQ organisations and causes.
Started by the Equality Network’s Scott Cuthbertson, the petition reads: "Chick Fil A has for years funded several anti-LGBT organisations and causes. Giving millions to organisations that have opposed marriage and other rights for LGBT people.
"Lets ask Macdonalds not to support people who fund groups opposed to LGBT equality."
Speaking in an interview with PinkNews, Mr Cuthbertson said: “Scotland is a country with a proud LGBT legislative record, and the opening of such a restaurant isn’t compatible with that.
“Chick-fil-A need to learn that if they spend millions supporting anti-LGBT organisations that there should be consequences, consumers vote with their feet.
“Quietly sneaking into Scotland when you’ve been rejected south of the border won’t wash.”
A spokesperson for Chick-fil-A, Inc. responding to the petition, said: “We hope our guests in the U.K. will see that Chick-fil-A is a restaurant company focused on serving great food and hospitality, and does not have a social or political agenda.
"We are represented by more than 145,000 people from different backgrounds and beliefs, and we welcome everyone.”
In response to the claims about the money they have spent supporting anti-LGBT organisation, the spokesperson added: "In 2017 (the last reportable tax filing available), the Chick-fil-A Foundation gave $9.9 million in donations to communities across America.
"The sole focus of our donations was to support causes focused on youth and education. We are proud of the positive impact we are making in communities across America and have been transparent about our giving on our web site. To suggest our giving was done to support a political or non-inclusive agenda is inaccurate and misleading."
Opposition to the chain has also been backed by co-leader of the Scottish Greens, Patrick Harvie, who called on Holyrood MSPs to avoid the Macdonald Hotels chain following its decision to allow the chain to open.
Twitter users have also derided Macdonald Hotels, one person wrote: "Absolutely disgusting @MacdonaldHotels has welcomed anti equality @ChickfilA to Aviemore. Won’t use their hotels again."
While another added: "I’m disgusted. There is no place in Scotland for their brand of hatred."
Responding to the petition, Dillin Smith wrote: "To think any money spent at this business could financially contribute to the systematic suffering of the gay community is both terrifying and an affront to the values of our country. For shame to those who could possibly support this evil."
A spokesperson for Macdonald Aviemore Resort said: “We are pleased to have them [Chick-fil-A] 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.”