| AM Kitchens (Fife) Limited | Construction | 2026 |
| AO Elite Ltd | Arts & recreation | 2024 |
| Bayview Accounting Services Limited | Professional & scientific | 2014 |
| Bayview Carpets Ltd | Wholesale & retail | 2023 |
| Bedford Falls Equestrian Estate Ltd | Arts & recreation | 2026 |
| Bricksclothing Limited | Wholesale & retail | 2024 |
| Calgary Communications Limited | Professional & scientific | 2014 |
| Cruinn Consulting Ltd | Professional & scientific | 2018 |
| Cruinn Projects Limited | Administrative & support | 2022 |
| DCS Repairs & Maintenance Ltd | Manufacturing | 2025 |
| Denbeath Retail Ltd | Wholesale & retail | 2021 |
| Direct HGV Driver Training Ltd | Wholesale & retail | 2025 |
| DOE Sport (North) Limited | Construction | 1996 |
| East Dock Bar Ltd | Hospitality | 2026 |
| East Fife Football Club, Limited | Arts & recreation | 1911 |
| East Neuk 50 Cycle Route Community Interest Company | Construction | 2019 |
| EGA (Scotland) Ltd | Other services | 2025 |
| Fife Access Scaffolding Services Ltd | Construction | 2021 |
| Floorfit Fife Ltd | Construction | 2026 |
| Flora Fusion Ltd | Wholesale & retail | 2016 |
| G & G Recruitment Consultants Ltd | Administrative & support | 2016 |
| Groundhoggs Garden Services Ltd | Administrative & support | 2015 |
| International Testing Services Ltd | Professional & scientific | 2020 |
| JW Electrical (Fife) Ltd | Construction | 2022 |
| Kerr Miller Recruitment Ltd | Administrative & support | 2017 |
| Kingdom Carpets Limited | Wholesale & retail | 2010 |
| LF Access Limited | Other services | 2025 |
| Linrod Properties Ltd | Real estate | 2022 |
| LRS Pharmaesthetics Limited | Other services | 2026 |
| Lynch Bookkeeping & Payroll Ltd | Professional & scientific | 2019 |
| MADE4YOU Scotland Ltd | Administrative & support | 2024 |
| Mclean & SON Registered Plumbing, Heating & Renewable Engineers Ltd | Construction | 2023 |
| Millcastle EBS Limited | Professional & scientific | 2023 |
| Millcastle Limited | Financial services | 2023 |
| MMS Dynasty Holdings Ltd | Financial services | 2025 |
| Morespace Fife Ltd | Transport & logistics | 2019 |
| Myio Ltd | Professional & scientific | 2018 |
| Nonna's Pasta Ltd | Hospitality | 2026 |
| Northgate Training & Compliance Limited | Construction | 2025 |
| Prichard Developments Limited | Other services | 2015 |
| R.T. Stuart Bakers Limited | Manufacturing | 2017 |
| R.T. Stuart Limited | Manufacturing | 1941 |
| Rachel George Furniture Ltd | Wholesale & retail | 2025 |
| Sandy Cunningham Plumbing & Heating Limited | Construction | 2017 |
| Sea Harvest (Services) Ltd | Wholesale & retail | 2020 |
| Spey Inspection Ltd | Professional & scientific | 2025 |
| THE Tyre Store Accessories UK Ltd | Wholesale & retail | 2010 |
| Trade Bathroom Supplies Ltd | Wholesale & retail | 2023 |
| Wild Glen Florist Ltd | Wholesale & retail | 2023 |
| Wyse Water Solutions Ltd | Construction | 2021 |