Construction and characterization of BsGDH-CatIB variants and application as robust and highly active redox cofactor regeneration module for biocatalysis

Table 1 Protein production and productivity data of BsGDH wild type and BsGDH-CatIB variants

	CatIBs visibility (SDS) (Yes/No)	CatIBs visibility (Microscopy) (Yes/No)	Cells with CatIBs [%]	Pellet activity (Yes/No)	P_v Pellet (g L^-1d^-1)	Specific P_v Pellet (g L^-1d^-1 g_CatIB^-1)	P_v Supernatant (g L^-1d^-1)
BsGDH_WT	No	No	0	No	0	0	76.8
BsGDH-PT-TorA	No	No	0	No	0	0	1.9
BsGDH-PT-CBDCell	Yes	Yes	52	Yes	19.0	55.8	0.3
BsGDH-PT-3HAMP	Yes	Yes	6	Yes	12.6	48.3	0.8
BsGDH-PT-GFIL8	Yes	Yes	75	Yes	6.7	26.8	1.3
BsGDH-PT-L6KD	Yes	Yes	63	Yes	19.6	36.3	5.4
BsGDH-PT-18AWT	No	No	0	No	0	0	11.3
BsGDH-PT-TDoT	Yes	Yes	13	Yes	16.3	24.3	11.3
BsGDH-SG-ELK16	Yes	Yes	75	Yes	2.3	7.9	0
BsGDH-SG-TorA	No	Yes	32	No	0	0	24.7
BsGDH-SG-CBDCell	Yes	Yes	75	Yes	0.6	2.6	0
BsGDH-SG-3HAMP	Yes	Yes	16	Yes	1.0	3.2	3.6
BsGDH-SG-GFIL8	Yes	Yes	63	No	0	0	0
BsGDH-SG-18AWT	Yes	Yes	79	No	0	0	1.3
BsGDH-SG-TDoT	No	No	0	Yes	1.4	6.4	11.2

Pellet activity was tested by adding 200 mM glucose, 0.4 mM NAD⁺ and 40 mM TAE buffer (pH 7) to the CatIB pellet fraction after purification. Productivity was calculated after 20 min (pellet fraction), 12 min (supernatant; BsGDH-CatIBs) or 6 min (supernatant BsGDH_WT) reaction time. Microscopic images were taken after main cultivation (72 h) and analyzed regarding the number of cells that carry CatIBs (n ≥ 45).
P_V: volumetric productivity (g_NADH L⁻¹ d⁻¹ g_CatIB⁻¹)

ISSN: 1475-2859