| Durie Farms Limited | Agriculture & fishing | 2007 |
| Dusted Limited | Administrative & support | 2022 |
| E Scott Rentals Ltd | Real estate | 2024 |
| E&S Group Ltd | Professional & scientific | 2025 |
| E-Agora Limited | Wholesale & retail | 2026 |
| E-Descender Limited | Manufacturing | 2018 |
| E.G Developments Ltd | Construction | 2026 |
| Earlsferry Town Hall Limited | Other services | 2020 |
| East Fife Joinery Limited | Construction | 2014 |
| East Fife TV Limited | Information & communication | 2020 |
| East Links Wealth Limited | Financial services | 2026 |
| East Neuk Books Ltd | Wholesale & retail | 2014 |
| East Neuk Chef Limited | Hospitality | 2022 |
| East Neuk Literary Festival Limited | Arts & recreation | 2020 |
| East Neuk Properties Limited | Households | 2024 |
| East Neuk Spirit Company Ltd | Manufacturing | 2021 |
| East Scotland Joiners Ltd | Construction | 2025 |
| Eco Gleam Cleaning Limited | Administrative & support | 2021 |
| Eco Huts Fife Ltd | Hospitality | 2026 |
| Eco Your Home (Fife) Ltd | Energy | 2019 |
| Ecommerce by Aliraza Ltd | Wholesale & retail | 2026 |
| Ekkelkamp Contracting Operator Ltd | Construction | 2016 |
| Elder & Paton (Leven) Limited | Wholesale & retail | 2017 |
| Electric Bill Services Limited | Energy | 2017 |
| Elementx Group Limited | Financial services | 2026 |
| Elevett Global Limited | Other services | 2023 |
| Elie Escapes Limited | Hospitality | 2021 |
| Elie Golf Services Limited | Wholesale & retail | 2026 |
| Elie Leisure Company Ltd | Hospitality | 2016 |
| Elie Watersports Ltd | Arts & recreation | 2021 |
| Elite Resin Driveways and Landscaping Ltd | Construction | 2022 |
| Emaar Property Limited | Real estate | 2021 |
| Emma Gabellone Aesthetics Limited | Other services | 2017 |
| Energy Investments Limited | Professional & scientific | 2016 |
| Esme Headen Limited | Health & social care | 2014 |
| Euro Installations Ltd | Construction | 2025 |
| Ewan Archer Joinery Limited | Construction | 2024 |
| EX Electrical Limited | Manufacturing | 2011 |
| Excelsior Tree Surgery Limited | Agriculture & fishing | 2021 |
| Extra Energy Savings Limited | Other services | 2023 |
| Fa-Latch Construction Ltd | Construction | 2022 |
| Families in Trauma Limited | Health & social care | 2015 |
| FCE Group Ltd | Financial services | 2016 |
| FCE Remik Ltd | Manufacturing | 1998 |
| Feat Trading Community Interest Company | Hospitality | 2012 |
| Feelmightea Ltd | Wholesale & retail | 2024 |
| Fence-N-Deck Ltd | Wholesale & retail | 2026 |
| Fenstone Limited | Wholesale & retail | 1999 |
| Field Clothing Ltd | Manufacturing | 2025 |
| Fife Accounting Services Limited | Professional & scientific | 2017 |
| Fife Care Limited | Health & social care | 2022 |
| Fife Coatings and Inspection Ltd | Construction | 2025 |
| Fife Contract Car Hire Limited | Transport & logistics | 1996 |
| Fife Employment Access Trust | Administrative & support | 2011 |
| Fife Heat Treatment, Sales&Rentals Ltd | Manufacturing | 2025 |
| Fife Life Ltd | Real estate | 2022 |
| Fife Orthopaedic Specialists and Associates - Fosa LLP | | 2009 |
| Fife Properties Ltd | Real estate | 2006 |
| Fiona Livingstone Fitness Limited | Arts & recreation | 2022 |
| First Class Barbers Scotland Ltd | Other services | 2023 |
| Fixed Wire Testing Scotland Ltd | Construction | 2025 |
| FL Services (Aberdeen) Ltd | Manufacturing | 2026 |
| Flippin Properties (Fife) Limited | Construction | 2025 |
| Food Organisation Co Ltd | Hospitality | 2021 |
| Fortel Holdings Limited | Information & communication | 1996 |
| Fortel International Limited | Manufacturing | 1996 |
| Fortel Technology (UK) Limited | Information & communication | 1991 |
| Fortel Technology Limited | Professional & scientific | 1988 |
| Forth and Tide Properties Ltd | Real estate | 2025 |
| Forthview GAS Limited | Construction | 2023 |
| Fortune Garden Methil Limited | Hospitality | 2026 |
| Fortunest Ltd | Hospitality | 2024 |
| Fraser Properties (Fife) Ltd | Real estate | 2024 |
| Fraser Properties and Investments Ltd | Real estate | 2022 |
| Front Machinery Limited | Manufacturing | 2011 |
| FS1 Trade Limited | Water & waste | 2019 |
| Fyml Limited | Manufacturing | 2014 |
| G & C Murray Limited | Wholesale & retail | 2021 |
| G Henderson & Sons Properties (Star) Limited | Real estate | 1994 |
| G&Kstyles Enterprises Limited | Wholesale & retail | 2026 |
| Gabellone Rigging Ltd | Manufacturing | 2016 |
| Gallda Consultants Limited | Professional & scientific | 2019 |
| Garraffo Catering Solutions Limited | Manufacturing | 2016 |
| Gary Graham Limited | Real estate | 2023 |
| GAS Cats Ltd | Construction | 2015 |
| GAT Plastering Ltd | Construction | 2018 |
| Gatterson Group Ltd | Health & social care | 2025 |
| GC Heating & Plumbing Limited | Construction | 2013 |
| GCM Haulage Ltd | Transport & logistics | 2025 |
| Gentle Touch Removals Ltd | Transport & logistics | 2023 |
| Gfrf Services Ltd | Health & social care | 2024 |
| Gg&Fm Limited | Real estate | 2020 |
| GGD Engineering Services Ltd | Professional & scientific | 2023 |
| Gibson Training and Care Limited | Health & social care | 2007 |
| Gingey's Crafting Corner Ltd | Wholesale & retail | 2025 |
| GJ Accounting Services Ltd | Professional & scientific | 2021 |
| GJG Consulting Ltd | Professional & scientific | 2026 |
| Glassfibre Production Limited | Manufacturing | 2018 |
| Glenairlie Property Limited | Professional & scientific | 2026 |
| Gleneagles Holiday Park Limited | Hospitality | 2011 |