One-Step Enzymatic Synthesis of 1-Tuliposide A Using Tuliposide-Converting Enzyme

Nomura, Taiji; Ogita, Shinjiro; Kato, Yasuo

doi:10.1007/s12010-018-2903-3

Cited by 5 publications

(8 citation statements)

References 26 publications

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“…However, the HPLC t R of PosG obviously differed from that of PosD ( Figure S3). In the enzyme reaction of PosD by TgTCEA1, PosD was converted to PaA and 1-PosA ( Figure 3A), as we reported recently [16]. However, the enzyme reaction of PosG by TgTCEA1 resulted in the detectable formation only of an equimolar amount of PaA.…”

Section: Identification Of Posg In Tulip Bulbssupporting

confidence: 78%

“…To search for HPLC peaks arising from a novel Pos, crude methanolic extracts of tissues of the tulip cultivar 'Murasakizuisho', which has been used as a standard cultivar in our laboratory, were directly subjected to reaction with recombinant TgTCEA1 (T. gesneriana tuliposide A-converting enzyme 1), which catalyzes the removal of the acyl group at the C-6 position of Pos as Pa in a regiospecific manner [16]. Reversed-phase HPLC analysis of the reaction mixture revealed that a peak at retention time (t R ) 26 min, which was present in the reaction mixture without enzyme and did not correspond to a known Pos, disappeared after the TgTCEA1-catalyzed reaction ( Figure S1), suggesting that the t R 26 peak corresponds to a novel Pos, which we tentatively named 'PosG'.…”

Section: Identification Of Posg In Tulip Bulbsmentioning

confidence: 99%

“…However, at this stage, we could not find a clear HPLC peak corresponding to another product formed after the enzymatic removal of the C-6 acyl group from PosG (comparable to the 1-PosA formed from PosD). As the molar extinction coefficient of PosG (ε = 7940 at λ max 205 nm) was considerably smaller than that of PosD (ε = 17,800 at λ max 208 nm [16]), we predicted that the structure of PosG beyond the 6-PosA moiety lacks a π-conjugated system having strong ultraviolet (UV) absorbance. Then, the enzyme reaction of PosG by TgTCEA1 was performed at high substrate concentration (5 mM), and the reaction mixture was analyzed by HPLC with refractive index detection.…”

Section: Identification Of Posg In Tulip Bulbsmentioning

confidence: 99%

“…a This study. b Data taken from Nomura et al [16]. 1 H NMR and heteronuclear single quantum coherence spectroscopy (HSQC) spectra indicated the presence of one methine group (H-2″) and three methylene groups consisting of non-equivalent geminal protons (H-3″, H-4″, and H-5″) (Table 1, Figure S5A,D).…”

Section: Identification Of Posg In Tulip Bulbsmentioning

confidence: 99%

“…To identify high-performance liquid chromatography (HPLC) peaks corresponding to PosD and PosF in tulip tissue extracts, we adopted verification by reaction of the tissue extracts with tuliposide-converting enzymes (TCEs), based on our recent finding that PosD and PosF can serve as good substrates for TCEA and TCEB, respectively [16]; the corresponding Pos peaks disappeared from the HPLC chromatogram after reaction. The presence of a 4-hydroxy-2-methylenebutanoyl or 3,4-dihydroxy-2-methylenebutanoyl group at the C-6 position of d-glucose is crucial for being accepted as a substrate by TCEs, irrespective of the structural modification of the C-1 hydroxyl group of d-glucose [8].…”

Section: Introductionmentioning

confidence: 99%

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Identification of tuliposide G, a novel glucoside ester-type tuliposide, and its distribution in tulip

Nomura

Kato

2020

Zeitschrift Für Naturforschung C

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Tuliposides (Pos) are major defensive secondary metabolites in tulip (genus Tulipa), having 4-hydroxy-2-methylenebutanoyl and/or (3S)-3,4-dihydroxy-2-methylenebutanoyl groups at the C-1 and/or C-6 positions of d-glucose. The acyl group at the C-6 position is converted to antimicrobial lactones, tulipalins, by tuliposide-converting enzymes (TCEs). In the course of a survey of tulip tissue extracts to identify novel Pos, we found a minute high-performance liquid chromatography peak that disappeared following the action of a TCE, and whose retention time differed from those of known Pos. Spectroscopic analyses of the purified compound, as well as its enzymatic degradation products, revealed its structure as 5″-O-(6-O-(4′-hydroxy-2′-methylenebutanoyl))-β-d-glucopyranosyl-(2″R)-2″-hydroxymethyl-4″-butyrolactone, which is a novel glucoside ester-type Pos. We gave this compound the trivial name ‘tuliposide G’ (PosG). PosG accumulated in bulbs, at markedly lower levels than 6-PosA (the major Pos in bulbs), but was not found in any other tissues. Quantification of PosG in bulbs of 52 types of tulip, including 30 cultivars (Tulipa gesneriana) and 22 wild Tulipa spp., resulted in the detection of PosG in 28 cultivars, while PosG was present only in three wild species belonging to the subgenus Tulipa, the same subgenus to which tulip cultivars belong, suggesting the potential usefulness of PosG as a chemotaxonomic marker in tulip.

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Section: Identification Of Posg In Tulip Bulbssupporting

confidence: 78%

Section: Identification Of Posg In Tulip Bulbsmentioning

confidence: 99%

Section: Identification Of Posg In Tulip Bulbsmentioning

confidence: 99%

Section: Identification Of Posg In Tulip Bulbsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Identification of tuliposide G, a novel glucoside ester-type tuliposide, and its distribution in tulip

Nomura

Kato

2020

Zeitschrift Für Naturforschung C

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Protein Engineering for Enhanced Acyltransferase Activity, Substrate Scope, and Selectivity of the Mycobacterium smegmatis Acyltransferase MsAcT

et al. 2020

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The highly efficient and versatile acyltransferase MsAcT from Mycobacterium smegmatis catalyzes aqueous acyl transfer reactions, enabling applications in environmentally friendly processes and enzyme cascades. We rationally designed several variants with up to 30-fold increased acyl transfer to hydrolysis ratios while mostly retaining initial activity. Variants exhibiting broader acyl-donor substrate scope and higher or inverted enantioselectivity were also designed. Alterations of the catalytic His-Asp pair decreased the activation of hydrolytic water, thereby increasing acyl transfer to hydrolysis ratios. This study demonstrates that targeting the disruption of water networks and manipulating the activation of nucleophiles are promising strategies for engineering promiscuous acyltransferase activities.

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Isolation and identification of tuliposides D and F from tulip cultivars

Nomura

Ogita

Kato

2019

Zeitschrift Für Naturforschung C

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Abstract6-Tuliposides A (6-PosA) and B (6-PosB) are major defensive secondary metabolites in tulip cultivars (Tulipa gesneriana), having an acyl group at the C-6 position of d-glucose. Although some wild tulip species produce 1,6-diacyl-glucose type of Pos (PosD and PosF), as well as 6-PosA/B, they have not yet been isolated from tulip cultivars. Here, aiming at verifying the presence of PosD and PosF in tulip cultivars, tissue extracts of 25 cultivars were analyzed by high-performance liquid chromatography (HPLC). Although no HPLC peaks for PosD nor PosF were detected in most cultivars, we found two cultivars giving a minute HPLC peak for PosD and the other two cultivars giving that for PosF. PosD and PosF were then purified from petals of cultivar ‘Orca’ and from pistils of cultivar ‘Murasakizuisho’, respectively, and their identities were verified by spectroscopic analyses. This is the first report that substantiates the presence of 1,6-diacyl-glucose type of Pos in tulip cultivars.

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One-Step Enzymatic Synthesis of 1-Tuliposide A Using Tuliposide-Converting Enzyme

Cited by 5 publications

References 26 publications

Identification of tuliposide G, a novel glucoside ester-type tuliposide, and its distribution in tulip

Identification of tuliposide G, a novel glucoside ester-type tuliposide, and its distribution in tulip

Protein Engineering for Enhanced Acyltransferase Activity, Substrate Scope, and Selectivity of the Mycobacterium smegmatis Acyltransferase MsAcT

Isolation and identification of tuliposides D and F from tulip cultivars

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