Konjac glucomannan (KGM) is recognized as a safe material for its health-promoting benefits and thus widely used in various fields including pharmaceutical industry. In recent decades, the combination of collagen and KGM attracts more attentions for biomedical purpose, especially the hybrid films of collagen–KGM or collagen–KGM–polysaccharide. In this study, to further and deeply develop the intrinsic values of both collagen and KGM as biomaterials, a novel kind of composite hydrogel comprising collagen and KGM at a certain ratio was fabricated under mild conditions via fibrillogenesis process of the aqueous blends of collagen and KGM that experienced deacetylation simultaneously. The chemical composition, microcosmic architectures, swelling behavior, biodegradation and dynamic mechanic properties of such resulted composite hydrogels were systematically investigated. Biologic experiments, including cell culture in vitro and hypodermic implantation in vivo, were also conducted on these collagen/KGM composite hydrogels to evaluate their biologic performances. The relevant results prove that, based on collagen self-assembly behavior, this synthesis strategy is efficient to construct a composite hydrogel of collagen/KGM with improved mechanical properties, biodegradability, excellent biocompatibility and bioactivity, which are promising for potential biomedical applications such as tissue engineering and regenerative medicine.