Recombinant O-mannosylated protein production (PstS-1) from Mycobacterium tuberculosis in Pichia pastoris (Komagataella phaffii) as a tool to study tuberculosis infection

Table 1 Stoichiometric and kinetic parameters of P. pastoris growth and rPstS-1 production cultured in baffled shake flasks and in 1.0 L bioreactors

Parameter	Baffled shake flask	Bioreactor
Max O.D. 600 nm glycerol (A.U.)	26.5 ± 0.4	31.5 ± 2.2^b
Max O.D. 600 nm methanol (A.U.)	32.7 ± 0.8	47.0 ± 2.6^a
Max biomass glycerol (g/L)	8.8 ± 0.2 g/L	10.4 ± 0.9^b
Max biomass methanol (g/L)	10.8 ± 0.3 g/L	15.5 ± 0.9^a
μ glycerol (/h)	0.323 ± 0.025	0.286 ± 0.033
μ methanol (/h)	0.003 ± 0.001	0.006 ± 0.001^b
Y_x/s (g_DCW/g_Glyc)	0.69 ± 0.01	0.81 ± 0.03^b
Y_x/s (g_DCW/g_Meth)	0.051 ± 0.004	0.092 ± 0.004^a
q_s (g_Glyc/g_DCW h)	0.021 ± 0.004	0.034 ± 0.013
Supernatant total protein (mg/L)	127 ± 15	185 ± 9^a
Y_STP/DCW (mg _STP/g _DCW)	11.75 ± 1.13	11.92 ± 0.59
rPstS-1 (mg/L)	46 ± 5	46 ± 4
q_rPstS-1 (mg/L h)	0.639 ± 0.070	0.633 ± 0.054
q_p(rPstS-1) (mg_rPstS-1/g_DCW h)	0.059 ± 0.002	0.040 ± 0.004^a
Y_rPstS-1/x (mg_rPstS-1/g_DCW)	3.3 ± 0.01	4.0 ± 0.02^a

The mean and standard deviation for at least three biological replicates are shown
O.D., optical density; A.U., absorbance units; µ, specific growth rate; STP, supernatant total protein; DWC, dry cell weight; Y_x/s, total dry cell weight yield per glycerol; Y_x/s, total dry cell weight yield per methanol; Y_STP/DCW, STP yield per DCW; q_s, specific consumption rate of glycerol per DCW; q_PstS-1, volumetric productivity of rPstS-1; q_p(rPstS-1), specific productivity of rPstS-1; Y_rPstS-1/x, rPstS-1 yield per DCW. µ was calculated in first 8 h of culture
p value was calculated using t test by comparing the results of Shake flask VS Bioreactor
^ap < 0.005; ^b p < 0.05

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