| Elementary Leisure Ltd | Hospitality | 2025 |
| Elenita Properties Ltd | Real estate | 2026 |
| ELEV8 Mccoll Fitness Ltd | Arts & recreation | 2025 |
| Elise Mitchell Limited | Information & communication | 2024 |
| Elite Ironmongery Limited | Professional & scientific | 2015 |
| Elite-Landworx Ltd | Construction | 2025 |
| Elitevirtualsolutionsuk Ltd | Information & communication | 2025 |
| EM AI Ltd | Information & communication | 2026 |
| Emarketing Scotland Limited | Professional & scientific | 2014 |
| Emarketing Success Limited | Other services | 2015 |
| Ember Technology Holdings Limited | Financial services | 2020 |
| Ember Technology Ltd | Information & communication | 2016 |
| Empowered by Fitness Ltd | Arts & recreation | 2025 |
| EMS (Fife) Limited | Wholesale & retail | 2018 |
| ENM Enterprises Ltd | Wholesale & retail | 2026 |
| Enveraco Ltd | Professional & scientific | 2016 |
| Enviroworx Ltd. | Water & waste | 2014 |
| Epoint Embedded Computing Limited | Information & communication | 2008 |
| Equipco Investments Ltd | Professional & scientific | 2022 |
| Esme (Fife) Limited | Wholesale & retail | 2019 |
| Esosa Limited | Hospitality | 2025 |
| Euan Thomson Bespoke Ltd | Construction | 2023 |
| Eurocastors Limited | Manufacturing | 1968 |
| Eurospray Group Limited | Financial services | 2023 |
| Eurospray Limited | Construction | 1997 |
| Euthenia Corporate Ltd | Manufacturing | 2016 |
| Euthenia Recruitment Ltd | Administrative & support | 2017 |
| Eventrail Limited | Information & communication | 2017 |
| Evolved Energy Services Ltd | Construction | 2020 |
| Ewan Mackie Limited | Information & communication | 2015 |
| Ewen Property Holdings Limited | Real estate | 2023 |
| Ewen Property Ltd | Construction | 2008 |
| Ex-Spec Ltd | Professional & scientific | 2014 |
| Expert Auto Ltd | Wholesale & retail | 2020 |
| Eyzan Ltd | Information & communication | 2025 |
| EZ Boilers Limited | Construction | 2019 |
| F1 Developments Ltd | Construction | 2015 |
| F4 Holdings Ltd | Financial services | 2026 |
| F4 Lettings Ltd | Real estate | 2026 |
| Fabster Limited | Real estate | 2018 |
| Face IT Fife Aesthetics Ltd | Other services | 2021 |
| Fair Group (Fife) Ltd | Construction | 2025 |
| Falcon Fire Safety Ltd | Administrative & support | 2025 |
| FAR North Stays Limited | Real estate | 2026 |
| Fast Focus Logistics Limited | Transport & logistics | 2025 |
| Fast Recycling Ltd | Water & waste | 2020 |
| Fauji PK Ltd | Hospitality | 2020 |
| FDC Coatings Ltd | Construction | 2026 |
| Female Futsal Alliance CIC | Arts & recreation | 2025 |
| Fennes Jewellery Ltd | Wholesale & retail | 2018 |
| Ferrymuir Limited | Professional & scientific | 1995 |
| Fersa Ltd | Professional & scientific | 2001 |
| Fifab Holdings Limited | Manufacturing | 2017 |
| Fife & Kinross Contracts Ltd | Construction | 2016 |
| Fife & Kinross Joinery Ltd. | Construction | 2016 |
| Fife Adrenaline Sports Team Ltd | Education | 2016 |
| Fife Aluminium and Metal Company Ltd | Construction | 2016 |
| Fife Aluminium and Metal Installations Ltd | Construction | 2026 |
| Fife Auto Centre Ltd. | Wholesale & retail | 1999 |
| Fife Autotech Ltd | Wholesale & retail | 2020 |
| Fife Commercial Flooring Ltd | Construction | 2026 |
| Fife Cooperage Limited | Manufacturing | 2024 |
| Fife Developments Ltd | Construction | 2021 |
| Fife Female Football CIC | Arts & recreation | 2024 |
| Fife Free Cleaning Limited | Administrative & support | 2024 |
| Fife Gingerbread | Health & social care | 2009 |
| Fife Hydro Ltd | Wholesale & retail | 2025 |
| Fife IT Services Limited | Information & communication | 2015 |
| Fife Local Medical Committee Limited | Health & social care | 2010 |
| Fife LOG & Tree Services Ltd | Other services | 2014 |
| Fife Metal Recycling Limited | Wholesale & retail | 2025 |
| Fife Powdercoatings Limited | Wholesale & retail | 2016 |
| Fife Resource Solutions LLP | | 2013 |
| Fife Ropeaccess Ltd | Construction | 2025 |
| Fife Services Ltd | Construction | 2019 |
| Fife Shopmobility Limited | Other services | 1990 |
| Fife Shopping & Support Services Ltd | Health & social care | 2010 |
| Fife Shutters Limited | Manufacturing | 2002 |
| Fife Society for the Blind | Health & social care | 1996 |
| Fife Storage Solutions Ltd | Transport & logistics | 2024 |
| Fife Van Repair Centre Limited | Wholesale & retail | 2017 |
| Fife Vans and Trucks Limited | Wholesale & retail | 2009 |
| Fife Voluntary Action | Administrative & support | 2000 |
| Fife Windows & Doors Limited | Manufacturing | 2003 |
| Fife Women's AID | Hospitality | 2007 |
| Fifefoods Limited | Hospitality | 2026 |
| FIG Tree Hotel Ltd | Hospitality | 2025 |
| Finror Limited | Manufacturing | 2017 |
| Firebelly 83 Ltd | Wholesale & retail | 2025 |
| Firework CJR Limited | Administrative & support | 2025 |
| First Endeavour Holdings Limited | Financial services | 2022 |
| First for Homes Limited | Real estate | 2015 |
| Fishing Lures Ltd | Wholesale & retail | 2020 |
| FIX Auto Garage Ltd | Wholesale & retail | 2025 |
| FIX4YOU Limited | Construction | 2026 |
| Fixing IT (Scotland) Ltd | Information & communication | 2018 |
| Flakes Dessert Parlour Ltd | Hospitality | 2019 |
| Fletcher Property Holdings Ltd | Real estate | 2025 |
| Flexcon Europe Limited | Manufacturing | 1996 |
| Flexel International Limited | Manufacturing | 1993 |