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Objectives
To explore an
l
-isoleucine (Ile)-induced biosensor for down-regulation of Ile synthesis pathway and enhancement of 4-hydroxyisoleucine (4-HIL) production in
Corynebacterium glutamicum
SN01.
Results
Four Ile-induced riboswitches (IleRSN) with different strength were screened from mutation library based on TPP riboswitch. Firstly, IleRSN were integrated into the chromosome of strain SN01 immediately upstream of
ilvA
gene. The 4-HIL titer of strains carrying P
tacM
-driven IleRS1 or IleRS3 (14.09 ± 1.07, 15.20 ± 0.93 g 4-HIL L
−1
) were similar with control strain S-
D5
I (15.73 ± 2.66 g 4-HIL L
−1
). Then, another copy of IleRS3-
ilvA
was integrated downstream of the chromosomal
cg0963
gene in SN01-derived strain D-RS with down-regulated
l
-lysine (Lys) biosynthesis. The Ile supply and 4-HIL titer increased in
ilvA
two-copy strains KIRSA-3-
D5
I and KIRSA-3-
9
I, and Ile concentration was maintained less than 35 mmol L
−1
under the control of IleRS3 during fermentation. The resulting strain KIRSA-3-
9
I produced 22.46 ± 0.96 g 4-HIL L
−1
.
Conclusion
The screened IleRS was effective in the dynamic down-regulation of Ile synthesis pathway in
C. glutamicum
, and IleRSN with different strength can be applied in various conditions.