We present computer-assisted construction of regular polygonal knots by origami. The construction is completed with an automated proof based on algebraic methods. Given a rectangular origami or a finite tape, of an adequate length, we can construct the simplest knot by three folds. The shape of the knot is made to be a regular pentagon if we fasten the knot tightly without distorting the tape. We perform the analysis of the knot fold further formally towards the automated construction and verification. In particular, we show the construction and proof of regular pentagonal and heptagonal knots. We employ a software tool called Eos (e-origami system), which incorporates the extension of Huzita's basic fold operations for construction, and Gröbner basis computation for proving. Our study yields more mathematical rigor and in-depth results about the polygonal knots.