Members of the Free Presbyterian Church sang hymns and passed out leaflets at the entrance of the Theatre At The Mill in Newtownabbey last night as theatre-goers made their way to the play.
The group assembled at around 7pm before being led by a banner to the theatre's entrance.
The protesters, led by Rev Brian McClung of the Newtownabbey Free Presbyterian Church, handed over a letter of protest to Newtownabbey Borough Council.
On Wednesday, the first night of the two-night sold-out show, not a single protester was present.
Last night the group explained this was because the play clashed with their weekly church meeting.
The play performed by the Reduced Shakespeare Company was thrust into the spotlight when members of the council's artistic board pulled it because of complaints from the DUP.
Following an outcry, the board then decided to put the play back on, a decision which was ratified by the full council.
Rev McClung described it as "perverse human nature" that the play had sold so many tickets since opposition was raised.
He said: "Primarily we are here tonight to stand up for the honour of God's word. We believe this play mocks the word of God.
"We are offended because people are mocking the scriptures and we are here to show our offense. Supposedly the council and society is to have regard for minorities, but they don't seem to have much regard for evangelical Christians when it comes to things like this."
Police maintained a small presence but left before the play began.
Audience member Glenn Mayes said it would have set a dangerous precedent if the DUP had been allowed to tell the public what they could and couldn't watch.
"If I want to come see it, I'll come see it," he said.
"Where that type of censorship stops I don't know, so I'm glad they put it back on and I hope that's the end of it, because anything can be found offensive."
Husband and wife Steve and Jill Laird admitted that they wouldn't have known about the play were it not for the row.
"They are entitled to protest if they want to," Steve said.
"I think they are entirely wrong and it's an indictment on our society in Northern Ireland that people take things so seriously and get offended so easily."
"We can do what we want. No one should be telling us that," Jill added.
Protester Norman Davidson said that by mocking the Lord and the Bible the play had opened the door for more anti-Christian groups to come into Northern Ireland and mock God as they please.
"I hope it will make people think about the direction that this country is heading without God," he said.
"It needs a God in this society to cure all the ills that we see now, such as anti-social behaviour on the streets, drugs and alcohol."