In the third round, we addressed the solubility issue by narrowing down the input MSA to HLD subfamilies I-II and implementing stricter preprocessing steps to suppress the frequencies of indels. Moreover, we manually curated the sequences based on AlphaFold structural and MutCompute stability assessments. These steps were successful as the majority of the new designs were soluble. Interestingly, the non-curated AncDhaA13 showed good solubility and catalytic activity, despite our expected risky mutations P34V and L238F. Current computational tools are not particularly strong in predicting the epistasis effects. The curation step rescued poorly soluble AncDhaA17, which was crucial in designing AncDhaA16 and predicting the disruptive effect of the proline insertion on AncDhaA15 activity.