Light-inducible carotenoid production controlled by a MarR-type regulator in Corynebacterium glutamicum

Sumi, Satoru; Suzuki, Yuto; Matsuki, Tetsuro; Yamamoto, Takahiro; Tsuruta, Yudai; Mise, Kou; Kawamura, Takahiro; Ito, Yusuke; Shimada, Yuka; Watanabe, Eri; Watanabe, Shoko; Toriyabe, Minami; Shiratori, Hatsumi; Uéda, Kenji; Takano, Hideaki

doi:10.1038/s41598-019-49384-7

Cited by 26 publications

(19 citation statements)

References 46 publications

(66 reference statements)

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“…Decaprenoxanthin production in Clip185 was induced under light (Figure 13A, right). Decaprenoxanthin production is strongly induced under light in C. glutamicum as stated in published literature 19 in contrast to that in Clip185 (Figure 13A; Figure 13B). K. rhizophila produces decaprenoxanthin constitutively irrespective of light conditions (Figure 13C).…”

Section: Light-regulation Of Decaprenoxanthin Production In Clip185supporting

confidence: 61%

“…[16][17][18] In many Actinobacteria, decaprenoxanthin biosynthesis is not light-regulated, but in some Actinobacteria decaprenoxanthin biosynthesis is strictly induced under light. 19 Light induces decaprenoxanthin biosynthesis in Clip185 like it does in the actinobacterium Corynebacterium glutamicum, 19 but the induction is weaker than that in C. glutamicum.…”

Section: Introductionmentioning

confidence: 98%

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Isolation and characterization of a heavy metal- and antibiotic-tolerant novel bacterial strain from a contaminated culture plate of Chlamydomonas reinhardtii, a green micro-alga.

et al. 2021

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Background: Chlamydomonas reinhardtii, a green micro-alga, is normally cultured in laboratories in Tris-Acetate Phosphate (TAP), a medium which contains acetate as the sole carbon source. Acetate in TAP can lead to occasional bacterial and fungal contamination. We isolated a yellow-pigmented bacterium from a Chlamydomonas TAP plate. It was named Clip185 based on the Chlamydomonas strain plate it was isolated from. In this article we present our work on the isolation, taxonomic identification and physiological and biochemical characterizations of Clip185. Methods: We measured sensitivities of Clip185 to five antibiotics and performed standard microbiological tests to characterize it. We partially sequenced the 16S rRNA gene of Clip185. We identified the yellow pigment of Clip185 by spectrophotometric analyses. We tested tolerance of Clip185 to six heavy metals by monitoring its growth on Lysogeny Broth (LB) media plates containing 0.5 mM -10 mM concentrations of six different heavy metals. Results: Clip185 is an aerobic, gram-positive rod, oxidase-negative, mesophilic, alpha-hemolytic bacterium. It can ferment glucose, sucrose and mannitol. It is starch hydrolysis-positive. It is very sensitive to vancomycin but resistant to penicillin and other bacterial cell membrane- and protein synthesis-disrupting antibiotics. Clip185 produces a C50 carotenoid, decaprenoxanthin, which is a powerful anti-oxidant with a commercial demand. Decaprenoxanthin production is induced in Clip185 under light. NCBI-BLAST analyses of the partial 16S rRNA gene sequence of Clip185 revealed a 99% sequence identity to that of Microbacterium binotii strain PK1-12M and Microbacterium sp. strain MDP6. Clip185 is able to tolerate toxic concentrations of six heavy metals. Conclusions: Our results show that Clip185 belongs to the genus Microbacterium. In the future, whole genome sequencing of Clip185 will clarify if Clip185 is a new Microbacterium species or a novel strain of Microbacterium binotii, and will reveal its genes involved in antibiotic-resistance, heavy-metal tolerance and regulation of decaprenoxanthin biosynthesis.

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Section: Light-regulation Of Decaprenoxanthin Production In Clip185supporting

confidence: 61%

Section: Introductionmentioning

confidence: 98%

Isolation and characterization of a heavy metal- and antibiotic-tolerant novel bacterial strain from a contaminated culture plate of Chlamydomonas reinhardtii, a green micro-alga.

et al. 2021

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“…C. glutamicum and C. callunae are close relatives, synthesizing the C50 carotenoid decaprenoxanthin and its glycosides as pigments [ 25 , 27 ]. P. nicotinovorans is pigmented most probably due to the accumulation of carotenoids [ 35 ]. M. luteus is a yellow-pigmented bacterium due to the accumulation of the C50 carotenoid sarcinaxanthin and its glycosides [ 53 ].…”

Section: Discussionmentioning

confidence: 99%

“…CrtR has also been associated with the light-dependent regulation of carotenogenesis, although the mechanism remains unclear [ 35 ]. CrtR homologs are found mainly in actinobacteria, and crtR genes often cluster with carotenoid biosynthetic genes and/or a mmpL -like transporter gene [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

Corynebacterium glutamicum CrtR and Its Orthologs in Actinobacteria: Conserved Function and Application as Genetically Encoded Biosensor for Detection of Geranylgeranyl Pyrophosphate

Henke

Austermeier

Grothaus

et al. 2020

IJMS

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Carotenoid biosynthesis in Corynebacteriumglutamicum is controlled by the MarR-type regulator CrtR, which represses transcription of the promoter of the crt operon (PcrtE) and of its own gene (PcrtR). Geranylgeranyl pyrophosphate (GGPP), and to a lesser extent other isoprenoid pyrophosphates, interfere with the binding of CrtR to its target DNA in vitro, suggesting they act as inducers of carotenoid biosynthesis. CrtR homologs are encoded in the genomes of many other actinobacteria. In order to determine if and to what extent the function of CrtR, as a metabolite-dependent transcriptional repressor of carotenoid biosynthesis genes responding to GGPP, is conserved among actinobacteria, five CrtR orthologs were characterized in more detail. EMSA assays showed that the CrtR orthologs from Corynebacteriumcallunae, Acidipropionibacteriumjensenii, Paenarthrobacternicotinovorans, Micrococcusluteus and Pseudarthrobacterchlorophenolicus bound to the intergenic region between their own gene and the divergently oriented gene, and that GGPP inhibited these interactions. In turn, the CrtR protein from C. glutamicum bound to DNA regions upstream of the orthologous crtR genes that contained a 15 bp DNA sequence motif conserved between the tested bacteria. Moreover, the CrtR orthologs functioned in C. glutamicum in vivo at least partially, as they complemented the defects in the pigmentation and expression of a PcrtE_gfpuv transcriptional fusion that were observed in a crtR deletion mutant to varying degrees. Subsequently, the utility of the PcrtE_gfpuv transcriptional fusion and chromosomally encoded CrtR from C. glutamicum as genetically encoded biosensor for GGPP was studied. Combined FACS and LC-MS analysis demonstrated a correlation between the sensor fluorescent signal and the intracellular GGPP concentration, and allowed us to monitor intracellular GGPP concentrations during growth and differentiate between strains engineered to accumulate GGPP at different concentrations.

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“…Naturally, C. glutamicum contains the glycosylated C50 carotenoid decaprenoxanthin as a yellow pigment and, correspondingly, carotenoid biosynthetic genes of the non-mevalonate pathway, involving isopentenyl pyrophosphate as a central intermediate [104]. Native carotenoid biosynthesis is controlled by the GGPP-responsive transcriptional repressor CrtR [108] and is increased via CrtR upon exposure to light [109].…”

Section: Carotenoidsmentioning

confidence: 99%

Advances in metabolic engineering of Corynebacterium glutamicum to produce high-value active ingredients for food, feed, human health, and well-being

Wolf

Becker

Tsuge

et al. 2021

Essays in Biochemistry

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The soil microbe Corynebacterium glutamicum is a leading workhorse in industrial biotechnology and has become famous for its power to synthetise amino acids and a range of bulk chemicals at high titre and yield. The product portfolio of the microbe is continuously expanding. Moreover, metabolically engineered strains of C. glutamicum produce more than 30 high value active ingredients, including signature molecules of raspberry, savoury, and orange flavours, sun blockers, anti-ageing sugars, and polymers for regenerative medicine. Herein, we highlight recent advances in engineering of the microbe into novel cell factories that overproduce these precious molecules from pioneering proofs-of-concept up to industrial productivity.

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Light-inducible carotenoid production controlled by a MarR-type regulator in Corynebacterium glutamicum

Cited by 26 publications

References 46 publications

Isolation and characterization of a heavy metal- and antibiotic-tolerant novel bacterial strain from a contaminated culture plate of Chlamydomonas reinhardtii, a green micro-alga.

Isolation and characterization of a heavy metal- and antibiotic-tolerant novel bacterial strain from a contaminated culture plate of Chlamydomonas reinhardtii, a green micro-alga.

Corynebacterium glutamicum CrtR and Its Orthologs in Actinobacteria: Conserved Function and Application as Genetically Encoded Biosensor for Detection of Geranylgeranyl Pyrophosphate

Advances in metabolic engineering of Corynebacterium glutamicum to produce high-value active ingredients for food, feed, human health, and well-being

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