Lotus seeds represent a significant economic crop and are abundant in starch. To further enhance their application value, this study investigates the structural characteristics of lotus seed starch (LS) under the combined influence of pullulanase and high hydrostatic pressure (HHP). Pullulanase increased amylose content from 39.80 % to 72.26 %, and HHP increased amylose content further. LS crystals changed from C-type to B-type, and the ordered structure of LS was destroyed by enzymatic hydrolysis, and amylose single helix and partial double helix structure were formed. At low concentration, lotus seed amylose single helix tends to form amylose double helix structure with itself. At high concentrations, they tend to aggregate, forming a network structure with large surface area and loose order. HHP destroys the double helix structure of amylose, resulting in the decrease of starch crystallinity. These findings provide new insights into improving the processing properties and application range of lotus seed starch.