The invention discloses a strain of Acinetobacter and use thereof in the production of chiral 3-cyclohexene-1-carboxylic acid. Its taxonomic name is Acinetobacter sp., which is deposited on Jan. 21, 2019 at the China General Microbiological Culture Collection Center, under accession number CGMCC No. 17220. Using the Acinetobacter strain of the invention to produce chiral methyl 3-cyclohexene-1-carboxylate, the resulting methyl (S)-3-cyclohexene-1-carboxylate has an optical purity of 99% or more, and the catalyst has good stability, mild reaction condition and can withstand high concentrations of substrate and product. Using the resolution process of the invention, (S)-3-cyclohexene-1-carboxylic acid with high optical purity and high concentration can be simply and efficiently obtained, and the process is energy-saving and environmentally friendly, and the high-concentration of product is beneficial to downstream product recovery process. The invention provides an efficient method for production of (S)-3-cyclohexene-1-carboxylic acid, and has a good industrial application prospect.