Table 2.

Summary of yield, stability, and bioactivity of cloacaenodin variants. For yield, +++ signifies made at near wild-type levels, ++ signifies made at ~40-60% wild-type levels, + signifies made at ≤10% wild-type levels, all as detected by absorbance at UV-215 measured from HPLC. L signifies only detected via LC-MS. Stability was judged by ratio of threaded-to-unthreaded peptide in supernatant extract after purification, judged by LC-MS. N/A stands for not applicable. Variants that were not tested for bioactivity are listed as n.d. for no data.

Cloacaenodin Variant	Yield Relative to WT	Stability Relative to WT	Bioactivity Relative to WT
F22W	++	Comparable	Comparable
Y23W	+	More	n.d.
S24G	+++	Less	Abolished
S24T	++	Comparable	Less
S24C	++	Comparable	Less
S24A	++	Comparable	Comparable
S24Y	++	Comparable	Comparable
10-membered ring	L	Less	n.d.
G1A	Not tolerated	N/A	N/A
E9D	Not tolerated	N/A	N/A
P8A	++	Comparable	n.d.
V4P	L	More	n.d.
Y10A	+	Comparable	Much less