Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli

Misawa, Norihiko; Nakagawa, Masaya; Kobayashi, Kazuo; Yamano, Shigeyuki; Izawa, Yuko; Nakamura, Katsumi; Harashima, Keiji

doi:10.1128/jb.172.12.6704-6712.1990

Cited by 548 publications

(401 citation statements)

References 27 publications

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“…First, CRT-I type of phytoene desaturases have been cloned from the bacteria Rhodobacter capsulatus [13], Erwinia uredovora and E. herbicola (crtl genes) [14,15] and from the fungus Neurospora crassa (aLl gene) [16] and sequenced. Then, the genes encoding phytoene desaturases from cyanobacteria [12,17], green algae and plants [11,18,19] have been isolated (pds genes).…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning and functional expression in E. coli of a novel plant enzyme mediating ξ‐carotene desaturation

et al. 1995

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~bstract We have cloned a cDNA from the plant Capsicum annuum which encodes a novel enzyme mediating the dehydrogenation of ~-carotene and neurosporene to lycopene when expressed in E. coli cells accumulating ~-carotene or neurosporene. this enzyme is unable to dehydrogenate either phytoene or lycopene. The deduced amino acid sequence suggests that this cDNA encodes a polypeptide whose mature size is ca. 59 kDa and which is synthesized as a precursor with a NH2-terminal extension resembling transit peptides for plastid targeting. Sequence comparison reveals 33-35% similarity with previously cloned plant or cyanobacterial phytoene desaturases. In contrast, only limited sequence similarity is found with a ~-carotene desaturase from the cyanobacterium Anabaena.

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Section: Discussionmentioning

confidence: 99%

Molecular cloning and functional expression in E. coli of a novel plant enzyme mediating ξ‐carotene desaturation

et al. 1995

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“…Briefly, Escherichia coli BL21 (DE3) cells (Novagen) were transformed with combinations of pACCAR25DcrtB and the expression constructs pETb-MPSY3, pETa-SPSY1, pETa-SPSY3, or empty vector pET23b (1). Carotenoids were extracted (Gallagher et al, 2004), resuspended in methanol, and subjected to HPLC analysis (Quinlan et al, 2007); zeaxanthin diglucoside was identified as before (Gallagher et al, 2004) and in comparison with literature data (Misawa et al, 1990).…”

Section: Functional Complementationmentioning

confidence: 99%

“…Maize PSY3 and sorghum PSY1 and PSY3 cDNAs were subcloned into the pET23 expression vector (Novagen) and transformed into Escherichia coli harboring pACCAR25DcrtB, which carries a bacterial carotenoid gene cluster missing the bacterial PSY gene crtB (Misawa et al, 1990). Cells produce the pathway end product, zeaxanthin diglucoside, only when a functional PSY enzyme is present; a peak corresponding to this end product is seen in an HPLC chromatogram of extracted carotenoids from positive control cells containing the entire carotenoid cluster, including bacterial PSY (pACCAR25; Fig.…”

Section: Psy3 Encodes a Functional Psymentioning

confidence: 99%

PSY3, a New Member of the Phytoene Synthase Gene Family Conserved in the Poaceae and Regulator of Abiotic Stress-Induced Root Carotenogenesis

Vallabhaneni²,

Wurtzel³

2007

Plant Physiology

240

216

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Abscisic acid (ABA) plays a vital role in mediating abiotic stress responses in plants. De novo ABA biosynthesis involves cleavage of carotenoid precursors by 9-cis-epoxycarotenoid dioxygenase (NCED), which is rate controlling in leaves and roots; however, additional bottlenecks in roots must be overcome, such as biosynthesis of upstream carotenoid precursors. Phytoene synthase (PSY) mediates the first committed step in carotenoid biosynthesis; with PSY3 described here, maize (Zea mays) and other members of the Poaceae have three paralogous genes, in contrast to only one in Arabidopsis thaliana. PSY gene duplication has led to subfunctionalization, with each paralog exhibiting differential gene expression. We showed that PSY3 encodes a functional enzyme for which maize transcript levels are regulated in response to abiotic stresses, drought, salt, and ABA. Drought-stressed roots showed elevated PSY3 transcripts and ABA, responses reversed by rehydration. By blocking root carotenoid biosynthesis with the maize y9 mutation, we demonstrated that PSY3 mRNA elevation correlates with carotenoid accumulation and that blocking carotenoid biosynthesis interferes with stress-induced ABA accumulation. In parallel, we observed elevated NCED transcripts and showed that, in contrast to dicots, root zeaxanthin epoxidase transcripts were unchanged. PSY3 was the only paralog for which transcripts were induced in roots and abiotic stress also affected leaf PSY2 transcript levels; PSY1 mRNA was not elevated in any tissues tested. Our results suggest that PSY3 expression influences root carotenogenesis and defines a potential bottleneck upstream of NCED; further examination of PSY3 in the grasses is of value for better understanding root-specific stress responses that impact plant yield.

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“…For example, for lycopene production, crtB, crtE, and crtI from Erwinia uredovora (now Pantoea ananatis) were cloned into E. coli ( Figure 4). [31] Varying the order of genes in the crt operon affected the titer of lycopene which is presumable due to the change in the expression level of each gene. [32] Choosing the gene sources was also important.…”

Section: Introduction Of Heterologous Genesmentioning

confidence: 99%

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

Park

Yang

et al. 2017

Adv. Biosys.

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Natural products have been attracting much interest around the world for their diverse applications, especially in drug and food industries. Plants have been a major source of many different natural products. However, plants are affected by weather and environmental conditions and their successful extraction is rather limited. Chemical synthesis is inefficient due to the complexity of their chemical structures involving enantioselectivity and regioselectivity. For these reasons, an alternative means of overproducing valuable natural products using microorganisms has emerged. In recent years, various metabolic engineering strategies have been developed for the production of natural products by microorganisms. Here, the strategies taken to produce natural products are reviewed. For convenience, natural products are classified into four main categories: terpenoids, phenylpropanoids, polyketides, and alkaloids. For each product category, the strategies for establishing and rewiring the metabolic network for heterologous natural product biosynthesis, systems approaches undertaken to optimize production hosts, and the strategies for fermentation optimization are reviewed. Taken together, metabolic engineering has enabled microorganisms to serve as a prominent platform for natural compounds production. This article examines both the conventional and novel strategies of metabolic engineering, providing general strategies for complex natural compound production through the development of robust microbial‐cell factories.

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Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli

Cited by 548 publications

References 27 publications

Molecular cloning and functional expression in E. coli of a novel plant enzyme mediating ξ‐carotene desaturation

Molecular cloning and functional expression in E. coli of a novel plant enzyme mediating ξ‐carotene desaturation

PSY3, a New Member of the Phytoene Synthase Gene Family Conserved in the Poaceae and Regulator of Abiotic Stress-Induced Root Carotenogenesis

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

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