The long-running Mr Spokes bike-hire business closed its doors at West Basin on Sunday after being paid out by the ACT government, which wants the land for residential and commercial development.
The nearby paddle-boat business settled in December, also paid out in a confidential deal.
Both businesses were bruised by their dealings with the Land Development Agency, with Mr Spokes owners Jillian Edwards and Martin Shanahan close to desperation last year over what they said then were bullying tactics from the agency.
But the couple said they were satisfied with the final deal. They closed on January 31, settling the deal on Monday.
The closure marks the end of an era, with Mr Spokes operating at the lake for as many as 35 years.
Mr Shanahan and Ms Edwards bought the business and the building in 2006 for $480,000 with a crown lease that was to run until 2027. Ms Edwards said the business had been operating for as many as 25 years before they bought it.
The couple were locked in a bitter dispute with the government for two years as the government sought to take back the area for its "city to the lake" project, developing West Basin for apartments, shops, a park and boardwalk, and plans also for hotels and a swimming pool. Work on the park begins this year and the first residential and commercial space is to be sold in the 2016-17 financial year.
After initially threatening to compulsorily acquire the land, then valuing the business at much less than the couple believed it was worth, the government called a halt in August 2015, telling them they could stay put while development went ahead around them.
The Land Development Agency wrote to the couple in August saying it "no longer needs to pursue the acquisition of your Crown lease". "A level of noise and other disruption will be unavoidable," the agency wrote.
Talks resumed and agreement was reached at the end of December. Details are confidential and neither party would reveal the payout figure, nor discuss the negotiations. The government has bought the building and the 250 bikes, along with the lease.
Ms Edwards said she was relieved and "a bit numb".
"We are very relieved. The uncertainty of what was happening, when combined with the stress of negotiations that had gone sour, that was awful, absolutely awful," she said.
Paddle-boat operator Jim Seears, who subleased the business from his brother Pat Seears, settled in December, handing over the business and about 45 canoes and paddle boats.
The boat hire had been on the site for 35 or 40 years, and he had operated it for 16 years, he said. He, too, found negotiations stressful and characterised recent months as "the big boys squashing the little boys".
"I didn't sleep for 12 months," he said. "The problems it caused with our health and the anxiety and the stress, so it's good to get that out of the way."
The end of the bike business leaves Canberra with no lakeside bike hire. Segways operate on the south side of the lake, and in January, a kayak, canoe and paddle-board hire business opened in Yarralumla.
Land Development Agency chief executive David Dawes said the agency was "exploring options to re-establish boat and bicycle hire operations".
The West Basin design included a boardwalk positioned just above water level, berthing facilities, bike lanes and pavilions suitable for the hire of bicycles, paddle boating and other watercraft, he said.
The agency has earmarked the sale of land for 200 residential dwellings, plus 5000 square metres of commercial space in the 2016-17 year, with similar amounts to follow in each of the following two years. Mr Dawes said the releases were subject to developing a promenade and the final alignment and timing of Parkes Way, which is to have a new road built over the top for access to the lakefront.