The clergyman argued rule by females was contrary to the Bible. The modern Church of Scotland has long rejected the infamous point of view, yet in some areas the Church still struggles with sexism.
Knox's stance would be strongly challenged by one woman who, having dealt with the poll-tax riots in the 1980s while working for a major local authority despite sympathising with the rioters, would undoubtedly get her point across.
Pauline Weibye is as near as the Kirk gets to having a chief executive and she'd relish the chance to tackle Knox.
The 60-year-old mother of two "feisty" grown-up daughters is one of a growing number of women in key roles in the Kirk since the so-called stained glass ceiling was finally broken only in 2004 with the first female Moderator, Dr Alison Elliot.
But the Church has also been forced to admit there are geographical pockets where there remains "painful" discrimination.
The current Moderator of the General Assembly of the Church of Scotland, the Right Reverend Lorna Hood, is only the third woman and the first parish minister to take on the ambassadorial one-year-role since the 16th century Reformation in Scotland.
Mrs Weibye, an "occasionally crusty" knitting fanatic, has a background in human resources and is striving to make the Kirk a "slicker" operation.
From her fifth-floor George Street office at the Church's administrative headquarters in the Scottish capital, Mrs Weibye looks out across Edinburgh and sees a snapshot of the wide-ranging work she oversees as the Kirk's first Secretary to the Council of Assembly.
The Church's pledge is to bring pastoral care to all in Scotland and beyond. It means more than just spiritual guidance and affinity. Running the Church is the same as any business with a £120m annual budget, says Mrs Weibye. But it also helps that she is Session Clerk at Craigmillar Park Church in Edinburgh.
She has spent her working life in public-sector management roles, joining local government soon after graduating from Edinburgh University.
In the newly-created position, Mrs Weibye ensures the KIrk nerve-centre runs smoothly as well as looking to revamp the image of the HQ, known as 121 after the building number, which she says is a title not always veiwed with positive connotations.
The Kirk does everything from carrying out so-called paupers' burials - where often the minister can be the only person present at the ceremony - to feeding the hungry through food banks.
The Kirk's presence is evident from the wealthy congregations, which subsidise the less well-off parishes, to the care homes, the safe havens for abuse victims and places where people with alcohol and drug dependencies can get help and support with no difficult questions asked.
The local governance aspect of Presbyterianism is emphasised in the rounded, involving structures of many churches that encourage participation.
The main decisions, though, reach national level. The General Assembly is also the highest court of the Kirk, consisting of around 400 ministers, 400 elders, and members of the diaconate, all representing the presbyteries.
Mrs Weibye said: "This is a new job, I'm the first person to hold it. It's not a top-down organisation. Sometimes it is said that it is chief executive authority, but I don't have the authority.
"It is not centralised, pyramidical organisation. It can sometimes be frustrating, but is a great challenge."
The Church has bounced back from bullying claims in its HQ but is still struggling with the issue of openly gay clergy.
At the General Assembly, the Kirk moved towards greater acceptance of gay clergy in the debate sparked in 2009 by the calling of openly gay minister the Reverend Scott Rennie to an Aberdeen church.
The move is still under consideration by the Church and will be for at least another year.
Mrs Weibye said: "You can't ignore the elephant in the room. The uncertainty around the issue of same-sex relationships is going to have an effect and it is harming."
But the feared schism over the issue has yet to materialise, with only 11 ministers having quit.
Fervour among hardline evangelicals has been said to be waning and many are seeking to push their case from with the Kirk as the debate continues.
She said in some areas congregation members wishing to stay with the Kirk have been left confused by those pushing to leave the Church.
"It's desperately sad," she says and she is concerned pastoral care may be affected.
Other women are also taking more key roles across the Kirk. The outspoken Reverend Sally Foster-Fulton speaks for the powerful Church and Society Council which takes a firm stance on issues affecting the country and its people, from next year's independence referendum to same-sex marriages.
Mrs Weibye, however, echoed the concerns of the current Moderator who surprised some in the Kirk when she spoke out again localised sexist attitides within the Church.
Mrs Hood said she was sure women who believed they were being denied recognition and positions of responsibility because of their gender would in the first instance raise this issue with their local Presbytery.
She called on women who did not feel their case received a fair hearing to come forward and said she was "taken aback and saddened to discover that in some pockets of Scotland there are women dedicated to the Lord and his Church who have experienced real discrimination".
She said she had "spent time with women still denied a place on Kirk sessions and local ruling bodies", adding: "There are women elders who, having moved from one area to another, find their ordination is called into question."
Mrs Weibye said while she had not experienced sexism in the church personally, she is hugely concerned about the issue.
She said: "The very real skills of women are not being recognised and used to the full in church committees and those women feel real frustration and pain."
She has also helped the Kirk bounce back from damaging bullying allegations made by those working in its HQ and stresses support is on hand for anyone with such concerns.
Brought up in Port Seton, her father was a church elder and she takes her name from her Norwegian husband whose seafaring family founded Norway House, the first members' club in Edinburgh for Norwegian armed forces and merchant navy personnel.
Of the poll-tax riots she experienced earlier in her working life, she said: "I was responsible for liaising with the police over security and also for media relations and found it challenging, partly because I had a lot of sympathy with the protesters. I was glad to move on from that role."
In her current role in an organisation run by committees and councils with no one person in charge, Mrs Weiby finds ways to keep tabs on what is going on.
The sea, a compelling draw to her, attracts her attention from time to time and on rare breaks in 121 she will steal a look across the Forth with binoculars at the ships in the Firth. When she gets home, she checks up on them.
"There's a website where you can track the ships all over the world. You know exactly where they are."
It seems she also knows excatly where the challenges in her day job lie.