辅因子（Cofactor）是指与酶（酵素）结合且在催化反应中必要的非蛋白质化合物。某些分子如水和部分常见的离子所扮演的角色和辅因子相当类似，但由于含量不受限制且普遍存在，因此不归类为辅因子。

一个不含辅因子的酶称为脱辅基酶（apoenzyme），脱辅基酶加上辅因子并产生完整作用时，称为全酶（holoenzyme）：

金属离子是常见的辅因子，这些金属离子反映在生物必须的微量元素名单当中。例如钙、镁、锰、铁、钴、镍、铜、锌与钼等。除了这些无机化学物之外，辅因子也包括一些有机物质，例如血红蛋白中的铁。另外有些维生素也可作为辅因子如维生素C；或是辅因子的前趋物，如维生素B₁。

A cofactor is a non-protein chemical compound that is required for the protein's biological activity. These proteins are commonly enzymes, and cofactors can be considered "helper molecules" that assist in biochemical transformations.

Cofactors can be subdivided into either one or more inorganic ions, or a complex organic or metalloorganic molecule called a coenzyme; most of which are derived from vitamins and from required organic nutrients in small amounts. A cofactor that is tightly or even covalently bound is termed a prosthetic group. Additionally, some sources also limit the use of the term "cofactor" to inorganic substances. An inactive enzyme without the cofactor is called an apoenzyme, while the complete enzyme with cofactor is called a holoenzyme.