The singer had been due on stage at the Theatre Royal in Norwich on Wednesday in Dvorak's Rusalka as part of the company's 2012 tour.
Glyndebourne said the performance had been cancelled out of respect to the singer's family and colleagues.
The company paid tribute to Poulton's "distinguished operatic career" and said he would be "enormously missed".
After studying at the Guildhall School of Music and Drama and the National Opera Studio, the singer began his professional career in the Glyndebourne chorus and performed central roles both in its festival and tours.
His recent projects included La Traviata with Opera Holland Park and the Royal Opera House, Tosca with the Scottish Opera, and Aida at the Royal Albert Hall.
International appearances included engagements with Netherlands Opera, Opera Zuid and Salzburger Landestheater.
"It is testament to Robert's generous character and amazing musicianship that he had such a distinguished operatic career, not just at Glyndebourne but also at many of the leading international houses," said Glyndebourne's general director David Pickard.
Poulton's management company also paid tribute following his death on Tuesday, saying his "immense contribution to the opera world will not be forgotten".
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