Dynamic control of 4-hydroxyisoleucine biosynthesis by multi-biosensor in Corynebacterium glutamicum

Lai, Wenmei; Shi, Feng; Tan, Shuyu; Liu, Haiyan; Li, Yongfu; Xiang, Youhe

doi:10.1007/s00253-022-12034-6

Cited by 10 publications

(9 citation statements)

References 47 publications

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“…The amino acids and 4-HIL concentrations were detected by Agilent 1260 HPLC detector equipped with a Thermo ODS-2 HYPER-SIL C18 column (250 mm × 4.6 mm, USA) using the ortho-phthalaldehyde precolumn derivatization method. The HPLC samples processed as described previously (Lai et al 2022).…”

Section: -Hil Fermentationmentioning

confidence: 99%

“…pBAD/arabinose), biosensors auto-induced by endogenous metabolites are more convenient and economical for regulation. In our previous work, several biosensors were applied to balance the cell growth and production in C. glutamicum SN01 to overproduce 4-HIL, including Lrp-P brnFE , L-lysine riboswitch (LysRS) and P ilvBNC attenuator (Lai et al 2022;Tan et al 2020).…”

Section: Dynamic Regulation Of Chromosomal Ilva Expression and 4-hil ...mentioning

confidence: 99%

“…The synthesis of Ile should be enhanced in Corynebacterium glutamicum to supply Ile for 4-HIL production. However, when Ile synthesis is extremely strengthened, IDO activity will be inhibited by excessive Ile, which resulted in the decrease of 4-HIL titer (Lai et al 2022; Tan et al 2020). In order to dynamically modulate the biosynthesis of Ile, the Ile synthesis pathway should be down-regulated when Ile is excessively accumulated.…”

Section: Introductionmentioning

confidence: 99%

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Exploring l-isoleucine riboswitches for enhancing 4-hydroxyisoleucine production in Corynebacterium glutamicum

et al. 2023

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ObjectivesTo 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. ResultsFour 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 . ConclusionThe 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.

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Section: -Hil Fermentationmentioning

confidence: 99%

Section: Dynamic Regulation Of Chromosomal Ilva Expression and 4-hil ...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Exploring l-isoleucine riboswitches for enhancing 4-hydroxyisoleucine production in Corynebacterium glutamicum

et al. 2023

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“…4-hydroxyphenylpyruvate (HPP) plays a pivotal role as a fundamental precursor in the biosynthesis of aromatic compounds, and the synthesis of numerous highvalue compounds can be achieved based on this premise [1,8] (Figure 1). Monitoring the amount of HPP in chassis cells can provide valuable support for the biosynthesis of these downstream compounds, and the development of biosensors for precursor compounds is beneficial for the synthesis of various downstream compounds [9]. In this regard, the development of biosensors is an important role.…”

Section: Introductionmentioning

confidence: 99%

“…Sci. 2024, 25, x FOR PEER REVIEW 2 of 17 support for the biosynthesis of these downstream compounds, and the development of biosensors for precursor compounds is beneficial for the synthesis of various downstream compounds [9]. In this regard, the development of biosensors is an important role.…”

Section: Introductionmentioning

confidence: 99%

Directed Evolution of 4-Hydroxyphenylpyruvate Biosensors Based on a Dual Selection System

Du,

Liang,

et al. 2024

IJMS

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Biosensors based on allosteric transcription factors have been widely used in synthetic biology. In this study, we utilized the Acinetobacter ADP1 transcription factor PobR to develop a biosensor activating the PpobA promoter when bound to its natural ligand, 4-hydroxybenzoic acid (4HB). To screen for PobR mutants responsive to 4-hydroxyphenylpyruvate(HPP), we developed a dual selection system in E. coli. The positive selection of this system was used to enrich PobR mutants that identified the required ligands. The following negative selection eliminated or weakened PobR mutants that still responded to 4HB. Directed evolution of the PobR library resulted in a variant where PobRW177R was 5.1 times more reactive to 4-hydroxyphenylpyruvate than PobRWT. Overall, we developed an efficient dual selection system for directed evolution of biosensors.

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Enhanced polysaccharide production through quorum sensing system in Cordyceps militaris

Qiao,

Chen,

Yang

2024

J Basic Microbiol

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This study aimed to enhance extracellular polysaccharide (EPS) production in Cordyceps militaris by constructing a quorum sensing (QS) system to regulate the expression of biosynthetic enzyme genes, including phosphoglucomutase, hexokinase, phosphomannomutase, polysaccharide synthase, and UDP‐glucose 4‐epimerase genes. The study found higher EPS concentrations in seven recombinant strains compared to the wild‐type C. militaris, indicating that the overexpression of key enzyme genes increased EPS production. Among them, the CM‐pgm‐2 strain exhibited the highest EPS production, reaching a concentration of 3.82 ± 0.26 g/L, which was 1.52 times higher than the amount produced by the wild C. militaris strain. Additionally, the regulatory effects of aromatic amino acids on the QS system of the CM‐pgm‐2 strain were investigated. Under the influence of 45 mg/L tryptophan, the EPS production in CM‐pgm‐2 reached 4.75 ± 0.20 g/L, representing a 1.90‐fold increase compared to wild C. militaris strains. This study provided an effective method for the large‐scale production of EPSs in C. militaris, and opened up new avenues for research into fungal QS mechanisms.

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Dynamic control of 4-hydroxyisoleucine biosynthesis by multi-biosensor in Corynebacterium glutamicum

Cited by 10 publications

References 47 publications

Exploring l-isoleucine riboswitches for enhancing 4-hydroxyisoleucine production in Corynebacterium glutamicum

Exploring l-isoleucine riboswitches for enhancing 4-hydroxyisoleucine production in Corynebacterium glutamicum

Directed Evolution of 4-Hydroxyphenylpyruvate Biosensors Based on a Dual Selection System

Enhanced polysaccharide production through quorum sensing system in Cordyceps militaris

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