Isolation, expression, and functional analysis of the geranylgeranyl pyrophosphate synthase (GGPPS) gene from Liriodendron tulipifera

Zhang, Chengge; Liu, HuanHuan; Zong, Yaxian; Tu, Zhonghua; Li, Huogen

doi:10.1016/j.plaphy.2021.06.052

Cited by 17 publications

(7 citation statements)

References 45 publications

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“…In conclusion, the expression of AlGGPPS and/or AlIDI, two key precursor synthase encoding genes in Aurantiochytrium, both improved the production of β-carotene (Figure 5b,d). The results were consistent with previous studies [8,[29][30][31]. Furthermore, the highest β-carotene yield was observed in the stain coordinately expressing AlGGPPS and AlIDI, indicating the additive effects of AlGGPPS and AlIDI on β-carotene overproduction.…”

Section: Discussionsupporting

confidence: 92%

“…Thereby, IDI and GGPPS define the availability of IPP, DMAPP and GGPP, functioning as the key regulatory nodes by directing metabolic flux to carotenoid biosynthesis. The two key precursor synthases have been derived from different species [24][25][26][27][28][29], and it has proved to be an effective strategy to strengthen DMAPP and GGPP supply through overexpression of IDI or/and GGPPS, diverting the metabolite flow towards carotenoid biosynthesis [8,[29][30][31].…”

Section: Introductionmentioning

confidence: 99%

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Identification and Functional Analysis of Two Novel Genes—Geranylgeranyl Pyrophosphate Synthase Gene (AlGGPPS) and Isopentenyl Pyrophosphate Isomerase Gene (AlIDI)—from Aurantiochytrium limacinum Significantly Enhance De Novo β-Carotene Biosynthesis in Escherichia coli

Shi

Chen

et al. 2023

Marine Drugs

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Precursor regulation has been an effective strategy to improve carotenoid production and the availability of novel precursor synthases facilitates engineering improvements. In this work, the putative geranylgeranyl pyrophosphate synthase encoding gene (AlGGPPS) and isopentenyl pyrophosphate isomerase encoding gene (AlIDI) from Aurantiochytrium limacinum MYA-1381 were isolated. We applied the excavated AlGGPPS and AlIDI to the de novo β-carotene biosynthetic pathway in Escherichia coli for functional identification and engineering application. Results showed that the two novel genes both functioned in the synthesis of β-carotene. Furthermore, AlGGPPS and AlIDI performed better than the original or endogenous one, with 39.7% and 80.9% increases in β-carotene production, respectively. Due to the coordinated expression of the 2 functional genes, β-carotene content of the modified carotenoid-producing E. coli accumulated a 2.99-fold yield of the initial EBIY strain in 12 h, reaching 10.99 mg/L in flask culture. This study helped to broaden current understanding of the carotenoid biosynthetic pathway in Aurantiochytrium and provided novel functional elements for carotenoid engineering improvements.

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Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Identification and Functional Analysis of Two Novel Genes—Geranylgeranyl Pyrophosphate Synthase Gene (AlGGPPS) and Isopentenyl Pyrophosphate Isomerase Gene (AlIDI)—from Aurantiochytrium limacinum Significantly Enhance De Novo β-Carotene Biosynthesis in Escherichia coli

Shi

Chen

et al. 2023

Marine Drugs

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“…The comparison of these amino acid sequences was shown in Figure 2B , with five highly conserved domains (from I to V), a result similar to that previously reported ( Chen et al, 1994 ). Two of these aspartic acid-rich motifs are located in domains II and V, respectively, where near the N-terminal of the amino acid sequence is known as FARM (first aspartic acid-rich motif) and near the C-terminal is known as SARM (second aspartic acid-rich motif) ( Zhang et al, 2021 ). In the previous crystal structures of prenyltransferase showed that the allylic substrate is bound in the middle of FARM and SARM via magnesium ions ( Aaron and Christianson, 2010 ).…”

Section: Resultsmentioning

confidence: 99%

Functional verification and characterization of a type-III geranylgeranyl diphosphate synthase gene from Sporobolomyces pararoseus NGR

Yan

Zhang

et al. 2022

Front. Microbiol.

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Carotenoids, a group of natural pigments, have strong antioxidant properties and act as precursors to vitamin A, which have garnered attention from industry and researchers. Sporobolomyces pararoseus represents a hyper-producer of carotenoids, mainly including β–carotene, torulene, and torularhodin. Geranylgeranyl diphosphate synthase (GGPPS) is regarded as a key enzyme in the carotenoid biosynthesis pathway. However, the precise nature of the gene encoding GGPPS in S. pararoseus has not been reported yet. Here, we cloned a cDNA copy of the GGPPS protein-encoding gene crtE from S. pararoseus NGR. The crtE full-length genomic DNA and cDNA are 1,722 and 1,134 bp, respectively, which consist of 9 exons and 8 introns. This gene encodes 377 amino acids protein with a predicted molecular mass of 42.59 kDa and a PI of 5.66. Identification of the crtE gene encoding a functional GGPPS was performed using heterologous complementation detection in Escherichia coli. In vitro enzymatic activity experiments showed that CrtE utilized farnesyl diphosphate (FPP) as an allylic substrate for the condensation reaction with isopentenyl diphosphate (IPP), generating more of the unique product GGPP compared to other allylic substrates. The predicted CrtE 3D-model was analyzed in comparison with yeast GGPPS. The condensation reaction occurs in the cavity of the subunit, and three bulky amino acids (Tyr110, Phe111, and His141) below the cavity prevent further extension of the product. Our findings provide a new source of genes for carotenoid genetic engineering.

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“…GGPP serves as a key precursor substrate of volatile and non-volatile terpenoids ( Beck et al, 2013 ). GGPPS encodes an important enzyme involved in the synthesis of volatile and non-volatile terpenoids, constituting a key node that regulates carbon flow in the isoprenoid pathway ( Zhang et al, 2021 ). Furthermore, based on Café pipeline analysis, terpene synthase (TPS) genes, including TPS2 , TPS4 , and TPS10 , were expanded in A. tsao-ko , and significantly over-represented in the monoterpenoid biosynthesis pathway.…”

Section: Resultsmentioning

confidence: 99%

“… Schematic diagram of MEP and MVA pathways ( Zhang et al, 2021 ). Each solid arrow represents a biosynthetic reaction step, and dashed arrows represent multiple-step reactions.…”

Section: Resultsmentioning

confidence: 99%

Genome Sequencing of Amomum tsao-ko Provides Novel Insight Into Its Volatile Component Biosynthesis

Sun

Yan

et al. 2022

Front. Plant Sci.

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As an important economic and medicinal crop, Amomum tsao-ko is rich in volatile oils and widely used in food additives, essential oils, and traditional Chinese medicine. However, the lack of the genome remains a limiting factor for understanding its medicinal properties at the molecular level. Here, based on 288.72 Gb of PacBio long reads and 105.45 Gb of Illumina paired-end short reads, we assembled a draft genome for A. tsao-ko (2.70 Gb in size, contig N50 of 2.45 Mb). Approximately 90.07% of the predicted genes were annotated in public databases. Based on comparative genomic analysis, genes involved in secondary metabolite biosynthesis, flavonoid metabolism, and terpenoid biosynthesis showed significant expansion. Notably, the DXS, GGPPS, and CYP450 genes, which participate in rate-limiting steps for terpenoid backbone biosynthesis and modification, may form the genetic basis for essential oil formation in A. tsao-ko. The assembled A. tsao-ko draft genome provides a valuable genetic resource for understanding the unique features of this plant and for further evolutionary and agronomic studies of Zingiberaceae species.

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Isolation, expression, and functional analysis of the geranylgeranyl pyrophosphate synthase (GGPPS) gene from Liriodendron tulipifera

Cited by 17 publications

References 45 publications

Identification and Functional Analysis of Two Novel Genes—Geranylgeranyl Pyrophosphate Synthase Gene (AlGGPPS) and Isopentenyl Pyrophosphate Isomerase Gene (AlIDI)—from Aurantiochytrium limacinum Significantly Enhance De Novo β-Carotene Biosynthesis in Escherichia coli

Identification and Functional Analysis of Two Novel Genes—Geranylgeranyl Pyrophosphate Synthase Gene (AlGGPPS) and Isopentenyl Pyrophosphate Isomerase Gene (AlIDI)—from Aurantiochytrium limacinum Significantly Enhance De Novo β-Carotene Biosynthesis in Escherichia coli

Functional verification and characterization of a type-III geranylgeranyl diphosphate synthase gene from Sporobolomyces pararoseus NGR

Genome Sequencing of Amomum tsao-ko Provides Novel Insight Into Its Volatile Component Biosynthesis

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