| 2BOX Group LP | | 2015 |
| 42DELIVERIES Ltd | Transport & logistics | 2026 |
| A & G King Timber Limited | Manufacturing | 1999 |
| A Carruthers Limited | Wholesale & retail | 2016 |
| A Hammond Ltd | Agriculture & fishing | 2023 |
| A Henderson Roofing Limited | Construction | 2020 |
| A.Q.F. Contractors Limited | Agriculture & fishing | 2012 |
| Ab.Built Ltd | Manufacturing | 2025 |
| ABC Early Learning Limited | Education | 2005 |
| Acon Limited | Construction | 2004 |
| Acorn 2 Limited | Extraterritorial | 2006 |
| ACTIV8 Lubricants Limited | Manufacturing | 2002 |
| AF Sloan | | 1986 |
| Agri Ignite Ltd | Information & communication | 2019 |
| AHV Cleaning Services Ltd | Administrative & support | 2025 |
| Alba Racing Limited | Arts & recreation | 2009 |
| Alexandra 1952 Limited | Extraterritorial | 2015 |
| Almagill Farm | | 1988 |
| Andrew James Windows & Conservatories Limited | Construction | 2014 |
| Annandale and Lochwood Estates LLP | | 2018 |
| Annandale Community Transport Services | Transport & logistics | 1999 |
| Annandale Timber Products Limited | Agriculture & fishing | 2008 |
| Archicraft Solutions Ltd | Construction | 2023 |
| Arla Foods GP Limited | Financial services | 2016 |
| Arla Foods Limited Partnership | | 2016 |
| Arla Foods Nairn Limited | Wholesale & retail | 1998 |
| AW Commercial Consultants Ltd | Professional & scientific | 2024 |
| B&G Properties (Lockerbie) Ltd | Real estate | 2025 |
| Bacol | Agriculture & fishing | 2025 |
| Bankhead of Tinwald Farm | | 2000 |
| Barefoot Engineering Design Ltd | Professional & scientific | 2020 |
| BAREFOOT7 Ltd | Manufacturing | 2020 |
| Barony Country Foods Ltd. | Manufacturing | 1998 |
| Beaulay Limited | Information & communication | 2023 |
| Bell Electrical Group Limited | Construction | 1994 |
| Bishopcleugh Pods Ltd | Hospitality | 2021 |
| Bistro 53 Limited | Hospitality | 2025 |
| BJN Services Limited | Health & social care | 2019 |
| Black Bull Gspc Limited | Hospitality | 2026 |
| Blindhillbush Farmers (NO 1) | | 2000 |
| Boden Ecology Ltd | Professional & scientific | 2025 |
| Bonshaw Grange Country Retreat Ltd | Hospitality | 2024 |
| Border Biofuels Limited | Wholesale & retail | 2013 |
| Border Fine Arts Company Limited | Extraterritorial | 1974 |
| Border Meats Limited | Agriculture & fishing | 2016 |
| Border Motorcycles Limited | Wholesale & retail | 2024 |
| Border Transport Services Ltd | Transport & logistics | 2014 |
| Border Valleys Healthcare Ltd | Health & social care | 2025 |
| Border Valleys Holdings Ltd | Real estate | 2025 |
| Borderlands Brewery Ltd | Manufacturing | 2025 |
| Boudicca Records LLP | | 2014 |
| Bracken Glen Ltd | Agriculture & fishing | 2023 |
| Braes Farm Kirtlebridge Limited | Agriculture & fishing | 2016 |
| Braw Estates Ltd | Real estate | 2025 |
| Broathill Farming Ltd | Agriculture & fishing | 2026 |
| Building Craftsmen (Dumfries) Limited | Construction | 1974 |
| Building Craftsmen (Yeovil) Limited | Construction | 1987 |
| Building Craftsmen Limited | Professional & scientific | 1998 |
| C G P Welding Limited | Manufacturing | 2008 |
| C.P. Porteous & Sons - Penlaw Farms | | 1985 |
| C.S.L Roofing Services Ltd | Construction | 2025 |
| Cairn Isle Property Company Limited | Professional & scientific | 1962 |
| Caledonia Construction Dumfries Ltd. | Construction | 2016 |
| Campbell Construction (Lockerbie) Ltd | Construction | 2021 |
| Cargen Tower Limited | Real estate | 2003 |
| Carruthers Bros. (Annan) Limited | Construction | 1975 |
| Castle Milk Nominees Limited | Real estate | 2000 |
| Castlehill Farms | | 1985 |
| Catenary Management Solutions Ltd | Real estate | 2017 |
| Chariots of Fire Equestrian Centre CIC | Education | 2016 |
| Charlesfield AD Limited | Energy | 2014 |
| Chevin Limited | Professional & scientific | 2007 |
| Coalnet LP | | 2016 |
| Corncockle Estate Limited | Hospitality | 2019 |
| Corncockle Farming Ltd | Agriculture & fishing | 2025 |
| Cowburn Farms | | 1991 |
| Cowen Hospitality Ltd | Administrative & support | 2025 |
| Crazy Critters Experience Limited | Administrative & support | 2017 |
| Creatomatic Ltd | Information & communication | 2012 |
| Cressfield House Limited | Hospitality | 2018 |
| Curling Stadium Europe Ltd | Arts & recreation | 2023 |
| D & L Timber (Holdings) Ltd | Financial services | 2023 |
| D & W Agriculture Limited | Agriculture & fishing | 2011 |
| D Cowan Joinery Ltd | Construction | 2018 |
| D J Kincaid Limited | Agriculture & fishing | 2012 |
| D&G Companion and Care Agency Limited | Health & social care | 2024 |
| D.S. Roofing Limited | Construction | 2006 |
| Daddysgym Ltd | Manufacturing | 2020 |
| Dalton Hook Farm Limited | Agriculture & fishing | 2015 |
| DAU DAU Ltd | Transport & logistics | 2026 |
| David Holmes Livestock Trading Ltd | Agriculture & fishing | 2018 |
| Davidholmeslockerbie Ltd | Agriculture & fishing | 2026 |
| Davidson Thorburn Consulting Limited | Professional & scientific | 2007 |
| Daviewalker Ltd | Transport & logistics | 2025 |
| Davrow Limited | Real estate | 2011 |
| Derek J Steen | | 2005 |
| DGB 2023 Ltd | Financial services | 2023 |
| DHJ Trading Limited | Administrative & support | 2019 |
| Dine Around Scotland Limited | Professional & scientific | 2008 |
| Dormont Distilling Limited | Manufacturing | 2017 |