Complete Pathway Elucidation and Heterologous Reconstitution of Rhodiola Salidroside Biosynthesis

Torrens-Spence, Michael P.; Pluskal, Tomáš; Li, Fu‐Shuang; Carballo, Valentina; Weng, Jing‐Ke

doi:10.1016/j.molp.2017.12.007

Cited by 103 publications

(131 citation statements)

References 69 publications

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“…We identified 11 contigs associated with partitiviruses from the transcriptome of Rhodiola rosea , which is a cold-tolerant perennial plant belonging to the family Crassulaceae used as traditional medicine (BioProject PRJNA398393) [34]. The sizes of 11 contigs ranged from 930 bp to 2,730 bp.…”

Section: Resultsmentioning

confidence: 99%

Identification of viruses belonging to the family Partitiviridae from plant transcriptomes

Cho

2020

Preprint

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1Viruses in the family Partitiviridae consist of non-enveloped viruses with bisegmented double-2 stranded RNA genomes. Viruses in this family have been identified from plants and fungi. In 3 this study, we identified several viruses belonging to the family Partitiviridae using plant 4 transcriptomes. From 11 different plant species, we identified a total of 74 RNA segments 5 representing 23 partitiviruses. Of 74 RNA segments, 28 RNA segments encode RNA-6 dependent RNA polymerases (RdRp) while 46 RNA segments encode coat proteins (CPs). 7According to ICTV demarcation for the family Partitiviridae, 25 RNAs encoding RdRp and 8 41 RNAs encoding CP were novel RNA segments. In addition, we identified eight RNA 9 segments (three for RdRp and five for CP) belonging to the known partitivruses. Taken together, 10 this study provides the largest number of partitiviruses from plant transcriptomes in a single 11 study. Viruses in the family Partitiviridae consist of non-enveloped viruses with bisegmented double-2 stranded (ds) RNA genomes [1]. Each RNA segment of the viruses in the family Partitiviridae 3 encodes a single protein including RNA-dependent RNA polymerase (RdRp) or coat protein 4 (CP). Members in the family Partitiviridae can be divided into five different genera according 5 to the host. For example, viruses in the genera Alphapartitivirus and Betapartitivirus are 6 identified from either plants or fungi while viruses in the genus Gammapartitivirus and 7 Deltapartitivirus are derived from only fungi and plants, respectively [1]. In addition, viruses 8 in the genus Cryspovirus are identified from protozoa. Transmission of partitiviruses is 9 occurred by seeds (plants), cell division and sporogenesis (fungi), and oocytes (protozoa) [1]. 10 To date, there are more than 45 partitivirus species. In particular, there are five deltapartitivirus 11 species infecting plants including Pepper cryptic virus 1 (PCV-1) [2], Pepper cryptic virus 2 12 (PCV-2), Fig cryptic virus (FCV) [3], Beet cryptic virus 2 (BCV2), and Beet cryptic virus 3 13 (BCV3) [4] according to International Committee on Taxonomy of Viruses (ICTV). 14 To identify novel partitiviruses with dsRNA genomes, dsRNA extraction followed by 15 next-generation sequencing (NGS) is an efficient approach. Based on those approaches, several 16 partitiviruses have been identified. For example, Melon necrotic spot virus was identified from 17 watermelon plant (Citrullus lanatus Thunb) using SOLiD NGS analysis [5]. Pittosporum 18 cryptic virus 1 was identified from an Italian pittosporum plant by NGS with extracted dsRNA 19 [6]. Similarly, application of NGS to extracted dsRNA has resulted in identification of 20 Trichoderma atroviride partitivirus 1 from the fungal species Trichoderma atroviride [7]. 21 Moreover, instead of traditional molecular cloning methods, in silico data analyses has 22 resulted in identification of diverse dsRNA viruses from expressed sequence tag (EST) 23 database [8] and plant transcriptomes [9-13]. Moreover, 120 mycoviruses infecting fungi have...

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Section: Resultsmentioning

confidence: 99%

Identification of viruses belonging to the family Partitiviridae from plant transcriptomes

Cho

2020

Preprint

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“…In plants of the Rhodiola genus, salidroside is synthesized from the aromatic amino acid L-Tyr in three enzymatic steps (Torrens-Spence et al, 2018). The involved glycosyltransferase, catalyzing the last step, uses uridine-diphosphate activated Glc (uridine diphosphate [UDP]-Glc) as donor for the sugar moiety.…”

Section: Pathway Reconstruction For Microbial Production Of Salidrosimentioning

confidence: 99%

Identification and Microbial Production of the Raspberry Phenol Salidroside that Is Active against Huntington’s Disease

et al. 2018

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Edible berries are considered to be among nature's treasure chests as they contain a large number of (poly)phenols with potentially health-promoting properties. However, as berries contain complex (poly)phenol mixtures, it is challenging to associate any interesting pharmacological activity with a single compound. Thus, identification of pharmacologically interesting phenols requires systematic analyses of berry extracts. Here, raspberry (Rubus idaeus, var Prestige) extracts were systematically analyzed to identify bioactive compounds against pathological processes of neurodegenerative diseases. Berry extracts were tested on different Saccharomyces cerevisiae strains expressing disease proteins associated with Alzheimer's, Parkinson's, or Huntington's disease, or amyotrophic lateral sclerosis. After identifying bioactivity against Huntington's disease, the extract was fractionated and the obtained fractions were tested in the yeast model, which revealed that salidroside, a glycosylated phenol, displayed significant bioactivity. Subsequently, a metabolic route to salidroside was reconstructed in S. cerevisiae and Corynebacterium glutamicum. The best-performing S. cerevisiae strain was capable of producing 2.1 mM (640 mg L 21 ) salidroside from Glc in shake flasks, whereas an engineered C. glutamicum strain could efficiently convert the precursor tyrosol to salidroside, accumulating up to 32 mM (9,700 mg L 21 ) salidroside in bioreactor cultivations (yield: 0.81 mol mol 21 ). Targeted yeast assays verified that salidroside produced by both organisms has the same positive effects as salidroside of natural origin.

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“…Leaf tissue of seventy-day-old E. grandis plants were harvested for total RNA extraction using Qiagen's RNeasy Mini Kit (Qiagen). Total RNA for A. thaliana, P. somniferum, C. roseus and R. rosea were produced as previously reported (65,66). First-strand cDNAs of were synthesized by RT-PCR using total RNA sample as template.…”

Section: Cdna Production and Molecular Cloningmentioning

confidence: 99%

“…A second multigene vector, containing the S. cerevisiae tyrosine-metabolism feedback-resistant mutants ARO4 K229L and ARO7 G141S , was additionally used to boost tyrosine flux into (S)norcoclaurine production in S. cerevisiae. This construct was built using the 2-µm, pTDH3, tTDH1, HIS3 multigene vector as previously described (66). (66).…”

Section: Cdna Production and Molecular Cloningmentioning

confidence: 99%

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Structural basis for divergent and convergent evolution of catalytic machineries in plant aromatic amino acid decarboxylase proteins

Torrens-Spence

Chiang

Smith

et al. 2018

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AbstractRadiation of the plant pyridoxal 5’-phosphate (PLP)-dependent aromatic L-amino acid decarboxylase (AAAD) family has yielded an array of paralogous enzymes exhibiting divergent substrate preferences and catalytic mechanisms. Plant AAADs catalyze either the decarboxylation or decarboxylation-dependent oxidative deamination of aromatic L-amino acids to produce aromatic monoamines or aromatic acetaldehydes, respectively. These compounds serve as key precursors for the biosynthesis of several important classes of plant natural products, including indole alkaloids, benzylisoquinoline alkaloids, hydroxycinnamic acid amides, phenylacetaldehyde-derived floral volatiles, and tyrosol derivatives. Here, we present the crystal structures of four functionally distinct plant AAAD paralogs. Through structural and functional analyses, we identify variable structural features of the substrate-binding pocket that underlie the divergent evolution of substrate selectivity toward indole, phenyl, or hydroxyphenyl amino acids in plant AAADs. Moreover, we describe two mechanistic classes of independently arising mutations in AAAD paralogs leading to the convergent evolution of the derived aldehyde synthase activity. Applying knowledge learned from this study, we successfully engineered a shortened benzylisoquinoline alkaloid pathway to produce (S)-norcoclaurine in yeast. This work highlights the pliability of the AAAD fold that allows change of substrate selectivity and access to alternative catalytic mechanisms with only a few mutations.SignificancePlants biosynthesize their own proteinogenic aromatic L-amino acids, namely L-phenylalanine, L-tyrosine and L-tryptophan, not only for building proteins but also for the production of a plethora of aromatic-amino-acid-derived natural products. Pyridoxal 5’-phosphate (PLP)-dependent aromatic L-amino acid decarboxylase (AAAD) family enzymes play important roles in channeling various aromatic L-amino acids into diverse downstream specialized metabolic pathways. Through comparative structural analysis of four functionally divergent plant AAAD proteins together with biochemical characterization and molecular dynamics simulations, we reveal the structural and mechanistic basis for the rich divergent and convergent evolutionary development within the plant AAAD family. Knowledge learned from this study aids our ability to engineer high-value aromatic-L-amino-acid-derived natural product biosynthesis in heterologous chassis organisms.

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Complete Pathway Elucidation and Heterologous Reconstitution of Rhodiola Salidroside Biosynthesis

Cited by 103 publications

References 69 publications

Identification of viruses belonging to the family Partitiviridae from plant transcriptomes

Identification of viruses belonging to the family Partitiviridae from plant transcriptomes

Identification and Microbial Production of the Raspberry Phenol Salidroside that Is Active against Huntington’s Disease

Structural basis for divergent and convergent evolution of catalytic machineries in plant aromatic amino acid decarboxylase proteins

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