Construction and comprehensive characterization of an EcLDCc-CatIB set—varying linkers and aggregation inducing tags

Table 1 Morphological and enzymatic data of the CatIB variants and controls

	CatIBs visibility [Yes/No]	Cells with CatIBs [%]	Ø CatIBs per cell [-]	Area_M of CatIBs [µm²]	Area_M of cells [µm²]	CatIB area per cell area [%]	Pellet activity [Yes/No]	Conversion after 3 min [%]	P_v [g L^− 1d^− 1]	Specific P_v [g L^− 1d^− 1 g_CatIB⁻¹]
Control_neg	No	0	0.00	0.00	2.72	0	No	0	0	0
LDC_WT	Yes	88	1.35	0.47	2.94	24	Yes	57^a 24^b	219^a 87^b	106^a
LDC-SG-TDoT	Yes	78	1.55	0.38	2.50	23	Yes	67	258	147
LDC-PT-TDoT	Yes	61	1.43	0.30	2.86	15	Yes	74	286	165
LDC-SG-18AWT	No	0	0.00	0.00	2.01	0	Yes	36	136	67
LDC-PT-18AWT	No	0	0.00	0.00	2.11	0	Yes	65	251	116
LDC-SG-L6KD	Yes	88	1.35	0.65	2.92	31	Yes	20	105	63
LDC-PT-L6KD	Yes	88	1.18	0.66	2.32	37	Yes	93	457	256
LDC-SG-GFIL8	Yes	72	1.44	0.49	2.76	28	Yes	41	156	83
LDC-PT-GFIL8	Yes	86	1.24	0.45	2.30	24	Yes	80	308	179
LDC-SG-3HAMP	Yes	82	1.78	0.48	2.81	32	Yes	59	227	115
LDC-PT-3HAMP	Yes	71	1.83	0.43	2.65	31	Yes	76	294	155

Pellet activity as well as conversion, after 3 min, was tested by adding 10 mM l-lysine, 0.1 mM PLP and 50 mM Kpi buffer (pH 7.2) to the CatIB pellet fraction after purification. Microscopic images were taken after main cultivation (72 h) and analyzed regarding the number of cells that carry CatIBs and the amount of CatIBs in the cells with CatIBs (n ≥ 111). Moreover, the area of cells and CatIBs were determined and the ratio of CatIB area per cell area (n = 100). Abbreviations: ^aPellet fraction, ^bSupernatant fraction, SN Supernatant, Area_MArea of the median, P_Vvolumetric productivity

ISSN: 1475-2859