During a engagement to Lichfield Cathedral on Monday, the King was heckled by a individual who asked him about his brother's connections with convicted sex offender Jeffrey Epstein.
The man asked how long the King had been aware about his brother's relationship with Epstein, and also raised doubts about whether he had instructed the authorities to "conceal" for Andrew.
Despite the heckling, there were throngs of supporters and cheers for the King, who was visiting the region for an event.
Prince Andrew's connections with Epstein have been under close examination in recent weeks. The royal, who gave up his royal duties earlier this month, has repeatedly denied any misconduct.
"How long have you known about Andrew and Epstein? Have you asked the authorities to cover up for him?"
The King did not respond but carried on greeting fans, some of whom chanted "Long live the King". One supporter can be overheard requesting him for a hug.
A female bystander outside the venue was also heard telling the protester to "leave" and to "be quiet".
The scandal over Prince Andrew's connections to Jeffrey Epstein has been revived by misconduct allegations repeated in Virginia Giuffre's recently released memoir.
In the memoir, Ms Giuffre claims she had sex with the royal on multiple times, including one instance with Epstein and "several other minors"
Prince Andrew, who reached a out-of-court agreement with Ms Giuffre in recent years, has consistently denied any wrongdoing. In a previous interview, he told a news program he did not recall meeting Ms Giuffre "ever", and that they "never had any sort of sexual contact."
But recently, fresh questions have been asked about how he is able to fund his way of life despite not being a working royal.
The revelation that he has only ever paid a token annual rent on his Windsor mansion has heightened the scrutiny over his living arrangements - even though he made significant contributions initially, including for renovations.
The political party said he should be summoned before members of parliament to give evidence about his lease of his residence.
But last Thursday, the government refused to give parliamentarians time to debate the royal's titles or his home, despite growing pressure for investigation.
Buckingham Palace has not issued a statement on whether the Duke of York might move out of his current residence - or where he might reside.
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