The addition of starch or skim milk to bovine serum albumin‐containing regeneration medium enhances the regeneration of both Bacillus amyloliquefaciens and B. subtilis protoplasts. With B. amyloliquefaciens starch could replace bovine serum albumin; with B. subtilis either starch or skim milk could be used instead of bovine serum albumin. The maximum regeneration frequency achieved was 10% with B. amyloliquefaciens and 290% with B. subtilis.