James Guthrie (December 5, 1792–March 3, 1869) was an American businessman and politician.

He was born in Nelson County, Kentucky, studied law and worked as a laywer until 1827 when he was elected to the Kentucky General Assembly . He served in Kentucky's lower house from 1827 to 1829, and in the upper house, the Kentucky Senate , from 1831 to 1840.

Guthrie served as president of Kentucky Constitutional Convention in 1849.

A city leader in Louisville, he invested in railroads, and served as president of the Louisville & Nashville Railroad and the Louisville and Portland Canal Company . He also helped found the University of Louisville and served as its first president.

Guthrie was appointed the 21st Secretary of the Treasury by President Franklin Pierce in 1853 and served for the entirety for Pierce's term.

He was also a member of the Peace Congress of 1861 held in Washington, D.C., to devise means to prevent the impending war

Guthrie also served in the United States Senate from 1865 through 1868, when he resigned due to ill health. He died the following year in Louisville and was buried in Cave Hill Cemetery.