Exploring the full natural diversity of single amino acid exchange reveals that 40–60% of BSLA positions improve organic solvents resistance

Bioresources and Bioprocessing

Table 2 Effect of each type of amino acid substitution on organic cosolvent resistance of BSLA

	AA substitutions from each type of AA in BSLA WT
	15 Aromatic AA in BSLA WT (300 substitutions)				79 Aliphatic AA in BSLA WT (1580 substitutions)				54 polar AA in BSLA WT (1080 substitutions)				33 charged AA in BSLA WT (660 substitutions)
	+ (44)	= (193)	− (36)	× (27)	+ (155)	= (843)	− (269)	× (313)	+ (108)	= (759)	− (136)	× (77)	+ (64)	= (417)	− (98)	× (81)
DMSO
Aromatic	7	20	8	0	9	14	17	19	8	14	24	19	22	12	21	15
Aliphatic	32	26	17	19	21	32	20	12	23	28	16	16	25	24	30	25
Polar	27	36	33	44	41	36	39	27	32	36	34	29	25	35	36	43
Charged	34	18	42	37	29	18	24	42	36	22	26	36	28	29	13	17
	+ (19)	= (195)	− (59)	× (27)	+ (60)	= (827)	− (380)	× (313)	+ (56)	= (780)	− (167)	× (77)	+ (24)	= (486)	− (69)	× (81)
DOx
Aromatic	5	22	3	0	17	13	15	19	16	13	22	19	17	14	19	15
Aliphatic	26	26	25	19	13	34	18	12	16	28	19	16	17	26	22	25
Polar	21	35	34	44	32	37	39	27	32	37	32	29	25	35	32	43
Charged	47	17	37	37	38	16	28	42	36	23	27	36	42	25	28	17
	+ (12)	= (211)	− (50)	× (27)	+ (69)	= (836)	− (362)	× (313)	+ (62)	= (751)	− (190)	× (77)	+ (38)	= (479)	− (62)	× (81)
TFE
Aromatic	8	19	6	0	23	12	17	19	18	15	14	19	16	15	16	15
Aliphatic	0	27	28	19	14	33	20	12	19	26	28	16	16	26	23	25
Polar	33	34	34	44	29	38	37	27	26	38	31	29	45	33	32	43
Charged	58	20	32	37	33	18	26	42	37	22	28	36	24	26	29	17

Twelve patterns per organic solvent are shown. The absolute number of AA substitutions per type is shown in brackets
Amino acid groups: aromatic (F, Y, and W), aliphatic (A, V, L, I, and G), charged (D, E, H, K, and R), and polar (C, M, P, S, T, N, and Q). For activity group definitions (+, =, −, ×), see text. Values are given as percentages for easier comparison, as BSLA amino acid composition is not evenly distributed (43.6% aliphatic, 29.8% polar, 18.2% charged and 8.3% aromatic AA)