Application of simple fed-batch technique to high-level secretory production of insulin precursor using Pichia pastoris with subsequent purification and conversion to human insulin

Table 1 Insulin precursor production with P. pastoris

Strain (copy#)	MeOH (%)	Induction time (h)	Final biomass (g L^-1)	Product^a (g L^-1) Culture supernatant	Product^b (g L^-1) Culture broth	Mass	Reference^c
Mut^+/s (6-8)	nd ^d	87	nd	1.5	nd	nd	[26]
Mut^s (nd)	1.0	72	80 to 90	0.22	nd	nd	[25]
Mut^s (11)	<0.1	110	89	0.3	nd	MS ^e	[24]
Mut^s (11)	1 to 0.5	136	109	0.25	nd	MS	[23]
Mut^s (nd)	nd	72	nd	3.6	1.47	nd	[27]
Mut⁺ (12)	0.8	96	nd	0.18	nd	nd	[28]
Mut⁺ (~2) ^f	0.2	88	59	3.84	3.075	MS	Present study

^a Concentrations were determined by reverse phase HPLC from clarified culture supernatant
^b Concentrations calculated considering the volumetric portion of biomass in the culture broth
^c Only references on secretory insulin precursor production with P. pastoris included containing respective quantitative data
^d nd; not determined
^e MS; mass spectrometry
^f Copy number not experimentally determined. Survival at zeocin concentrations up to 0.5 mg mL^-1 indicates a copy number of 2 [41].

ISSN: 1475-2859