Fatty acid metabolism not only affects cell membrane functionality but also correlates with the fruit quality and postharvest senescence. This study aims to assess the influence of calcium lactate immersion on lipid metabolism and senescence in ‘Jinfeng’ pear fruit during room temperature storage. The results demonstrated that calcium lactate effectively suppressed malondialdehyde content, respiratory rate, mass loss, and ethylene release, as well as phospholipase D (PLD), hydroperoxide lyase (HPL), lipoxygenase (LOX), lipase (LPS), alcohol acyltransferase (AAT), and alcohol dehydrogenase (ADH) activities in ‘Jinfeng’ pears. Additionally, calcium lactate resulted in an up-regulation of the gene expression levels associated with fatty acid biosynthesis, including PcACC, PcFabG, PcFabI, PcFabZ, PcKASII, PcACP, and PcFAD. Furthermore, there was an increase in the expression of genes involved in fatty acid elongation, such as PcKCS4, PcKCS6, PcKCS7, PcKCS10, PcLACS2, PcLACS4, PcLACS8, and PcCER. Conversely, it caused a down-regulation of the gene expression levels associated with fatty acid degradation, including PcLOX, PcAAT, PcADH, PcPLD, PcACD, PcECH, and PcACOX3. Simultaneously, calcium lactate treatment diminished the content of palmitic acid, while elevating the contents of linolenic acid, linoleic acid, and oleic acid. In summary, postharvest calcium lactate-immersed can efficaciously retard senescence of ‘Jinfeng’ pear fruit through regulating fatty acid lipid metabolism.