The American three-time world champion, who won the 100m by five thousandths of a second from Jamaican Kishane Thompson in 9.79sec, finished second in his semi-final behind Botswana's Letsile Tebogo.
Tebogo, also a finalist in the 100m, struck a psychological blow against Lyles, crossing the line in 19.96sec, while Lyles timed 20.08sec.
America's Kenny Bednaraek, the second-fastest in the world this year over the distance, clocked 20.00sec to edge out Alexander Ogando of the Dominican Republic in the first semi-final.
Erriyon Knighton of the US, the 20-year-old who has been a silver and bronze medallist over 200m at the past two world championships, qualified first from the third semi-final, clocking 20.09sec, ahead of Liberia's Joseph Fahnbulleh.
The two next-fastest qualifiers were Zimbabwe's Tapiwanashe Makarawu and his compatriot Makanakaishe Charamba, meaning defending Olympic champion Andre De Grasse does not qualify after a season in which the Canadian has been beset by injuries.
De Grasse said he knew it would be tough after he hurt his hamstring.
"After the 100m semis I didn't realise that when I came back the next day after the 200m first round I kind of re-aggravated a lingering issue that I have in my hamstring," he said.
"They gave me some meds for the pain, and so I just tried to come out here today and see what I could do. But it was going to be really tough to try to come away with a fast time, because of course these guys are running the Olympics."
South Africa's Wayde van Niekerk, the 400m Olympic champion at the 2016 Rio Games and world record holder at the distance, missed out on the 200m final, with his decision to step down to the shorter sprint failing to pay off.
"I'm a competitor, I won medals and I'm not happy with where I am at the moment," said Van Niekerk, 32.
Treble-chasing Lyles has the chance to be the standout star of the athletics programme in Paris.
Jamaican sprint star Shericka Jackson withdrew from the women's 100m and then pulled out of her 200m heat.
St Lucia's Julien Alfred, pursuing the women's sprint double, came second behind Gabby Thomas in the 200m after winning the shorter sprint.
Lyles, the Tokyo 2020 bronze medallist, has been dominant in his preferred 200m, winning the past three world titles.
He is the third-fastest half-lap runner in history, with his mark of 19.31sec set at the world championships in Oregon in 2022 ranking behind only Jamaica's Yohan Blake and Bolt's world record of 19.19sec.
Retired Jamaican sprint superstar Bolt did the 100m-200m sprint double at three consecutive Games -- Beijing 2008, London 2012 and Rio 2016, adding the 4x100m gold in London and Rio.
The first round of the sprint relay is on Thursday morning but Lyles is likely to be rested following his 200m semi-final.
The 200m final takes place at 1830 GMT on Thursday.
