Figure captions
Figure 1 - The schematic diagram of α-farnesene biosynthetic
pathways with NADPH and ATP regeneration pathway in Pichia
pastoris. The pathways of NADPH biosynthesis are shown in blue lines,
and the pathways of NADPH catabolism are shown in red lines. The
pathways of ATP biosynthesis are shown in green lines, and the pathways
of ATP catabolism are shown in pink lines. The key genes were listed in
ellipses. Abbreviations: IPP
(isopentenyl pyrophosphate), DMAPP
(dimethylallyl pyrophosphate), GPP
(geranyl pyrophosphate), FPP (farnesyl pyrophosphate).
Figure 2 – Screening the best enzyme in oxiPPP for α-farnesene
biosynthesis. (a) Overexpression of the key enzymes in oxiPPP affects
the intracellular NADPH level. (b)
The
effects of overexpression of the key enzymes in oxiPPP on α-farnesene
production after 72 h cultivation. The strain X33-30* was used as the
control (Grey bar), and the best strain is shown in red bar. These data
represent average values and standard deviations achieved from three
independent experiments.
Figure 3 – Optimization of the expression combination ways of
the enzymes in oxiPPP for α-farnesene biosynthesis. (a) The effects of
the different expression combination ways on the intracellular NADPH
level. (b) The effects of the different expression combination ways on
α-farnesene production after 72 h cultivation. The strain X33-30*S was
used as the control (Grey bar), and the best strain is shown in red bar.
These data represent average values and standard deviations achieved
from three independent experiments.
Figure 4 – Inactivation of PGI1 negatively affects α-farnesene
biosynthesis. (a) α-Farnesene titers. (b) Dry cell weight (DCW). (c)
The intracellular NADPH level at 24 h and 72 h. The strains X33-30* and
X33-31 were used as the control (Grey bar). These data represent average
values and standard deviations achieved from three independent
experiments.
Figure
5 - Optimization of the expression level of cPOS5 for α-farnesene
biosynthesis. (a) Dry cell weight (DCW). (b) The intracellular NADPH
level at 24 h and 72 h. (c) α-Farnesene titers and productivity. The
strain X33-31 was used as the control (Grey bar). These data represent
average values and standard deviations achieved from three independent
experiments.
Figure 6 –Overexpression of APRT to
increase ATP supply for α-farnesene production. (a) The cell growth of
strain X33-37 with overexpression of APRT. (b) The α-farnesene
production of strain X33-37 with overexpression of APRT. The strain
X33-35 without overexpression of APRT was used as the control. These
data represent average values and standard deviations achieved from
three independent experiments.
Figure 7 – Inactivation
of GPD1 to decrease the NADH consumption in shunt pathway. (a)
α-farnesene titers of strains
X33-37 and X33-38. (b) DCW of strains X33-37 and X33-38. (c) Glycerol
titers of strains X33-37 and X33-38. These data represent average values
and standard deviations achieved from three independent experiments.
Table
1. The main strains used in the study