Abstract Background and objectives In order to investigate the effect of wheat gluten on thermal‐vacuum packaged Chinese steamed bread (CSB) quality during storage at room temperature, the wheat gluten addition levels of 0%, 1%, and 2% were set to study the effect on the protein structure, water mobility, starch retrogradation properties, and texture properties. Findings With the addition of wheat gluten, the β‐sheet content was increased and random coil content of thermal‐vacuum packaged CSB decreased. Wheat gluten enhanced the water holding capacity of CSB and restricted water migration, resulting in a lower content of bound water and higher content of less strongly bound water and free water according to the low‐field nuclear magnetic resonance (NMR). With the increase of wheat gluten, the diffraction peak intensity and relative crystallinity reduced. The elasticity and cohesion were higher, and the hardness, gumminess, chewiness, and resilience were lower than those in the control. Conclusions The results of this study revealed that the addition of wheat gluten can effectively inhibit moisture dispersion and starch retrogradation to delay the staling of CSB during storage. Significance and novelty As a natural food additive, wheat gluten can not only optimize protein structure and enhance nutritional value but also extend the shelf life of CSB. Therefore, these findings will promote industrial development of CSB.