The effect of glycerol mixed substrate on the heterologous production of a Rhizopus oryzae lipase in Pichia pastoris system
► Mixed substrates are good strategy to increase protein production in Pichia pastoris. ► Methanol set-point and μ are the key parameters in the bioprocess production. ► Glycerol represses PAOX1 at μ highest than 0.04 h −1 in Mut s phenotype. ► Proteolytic activity was detected only when glycerol wa...
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Published in | Biochemical engineering journal Vol. 57; pp. 30 - 37 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Amsterdam
Elsevier B.V
15.11.2011
Elsevier |
Subjects | |
Online Access | Get full text |
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Summary: | ► Mixed substrates are good strategy to increase protein production in
Pichia pastoris. ► Methanol set-point and
μ are the key parameters in the bioprocess production. ► Glycerol represses
PAOX1 at μ highest than 0.04
h
−1 in Mut
s phenotype. ► Proteolytic activity was detected only when glycerol was used as co-substrate. ► Sorbitol was better co-substrate than glycerol for ROL production.
A recombinant
Rhizopus oryzae lipase producing Mut
s
Pichia pastoris strain was used as a model organism to study the effect of mixed substrates (glycerol and methanol) on the specific product productivity. Different fed-batch cultivations were performed under three constant specific growth rates (0.02, 0.05 and 0.1
h
−1), maintaining a constant methanol concentration of 2
g
l
−1.
At the lowest
μ tested (0.02
h
−1), the specific productivity was 1.23 and 1.61 fold higher and the specific methanol consumption rate (
q
sMeOH) was 3 and 3.5 fold higher than values obtained when
μ was 0.05 and 0.1
h
−1, respectively. This implies a relation between the
q
sMeOH and the specific productivity, yielding higher specific productivities whenever the consumption of methanol is higher. Although glycerol was maintained under limiting conditions in all
μ tested, when the relation between the
μ
Gly and
μ
MeOH was larger than 4, an important decrease on the maximal activity value was observed.
Finally, a comparison under the same conditions using glycerol or sorbitol as co-substrates was also performed, obtaining better specific productivity when sorbitol was used. In addition, protease activity was detected when glycerol was used as co-substrate. |
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Bibliography: | http://dx.doi.org/10.1016/j.bej.2011.08.004 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1369-703X 1873-295X |
DOI: | 10.1016/j.bej.2011.08.004 |