Kev Ntsuam Xyuas Ntawm Txoj Kev Biosynthetic Ntawm Acteoside

1.1 Phenylalanine Acteoside Metabolism Pathway

Tam sim no, dav dav biosynthetic muaj peev xwm metabolic modules ntawmActeosidetau mus txog qhov kev pom zoo, feem ntau suav nrog txoj hauv kev phenylalanine metabolic, txoj hauv kev dopamine / tyramine txoj hauv kev, thiab kev hloov pauv acyl downstream thiab glycosyl hloov pauv txoj hauv kev.

Cov kev tshawb fawb tau pom tias cov intermediates hauv synthesis ntawm Acteoside muaj xws li caffeic acid, caffeoyl-CoA, hydroxytyrosol thiab hydroxytyrosol glucoside [16-18]. Hauv vitro hydrolysis kev sim ua pov thawj tias Acteoside tau hydrolyzed rau hauv caffeic acid, de-rhamnosyl-Acteoside (mutongphenetoside A) thiab decaffeoyl-Acteoside nyob rau hauv kev ua ntawm glycoside hydrolase [19-20]. Kev tshawb fawb txog qhov cuam tshuam ntawm qhov kub thiab txias, lub teeb, pH thiab lwm yam ntawm kev ruaj ntseg ntawm Acteoside qhia tias Acteoside tuaj yeem degraded rau hauv caffeic acid thiab decaffeoyl-verbasin hauv kev daws [21]. Kev sim pub tsiaj txhu pom tau tias cov khoom siv hydrolyzed ntawm Acteoside hauv cov zis nas yog hydroxytyrosol thiab caffeic acid [22]. Nyob rau hauv txoj kev phenylalanine metabolic, phenylalanine yog thawj zaug hloov mus rau hauv cinnamic acid nyob rau hauv qhov kev txiav txim ntawm phenylalanine ammonia lyase (PAL), thiab cinnamic acid hloov mus rau phenylalanine nyob rau hauv qhov kev txiav txim ntawm cinnamic acid -4-hydroxylase (C4H). Lub nplhaib C4 yog tsim rau hauv p-coumaric acid, uas tom qab ntawd catalyzed los ntawm coumaric acid 3-hydroxylase (C3H) los tsim caffeic acid [3]. Tsis tas li ntawd, tyrosine ammonia lyase (TAL) muab tau los ntawm Saccharomyces radiata tuaj yeem hloov tyrosine mus rau p-coumaric acid, thiab p-coumaric acid tuaj yeem catalyzed los ntawm C3H los tsim caffeic acid, uas muab lub hauv paus rau heterologous synthesis ntawm caffeic acid. lub tswv yim tshiab [23].

Kom paub ntau ntxiv txog Cistanche Acteoside Function

1.2 Dopamine txoj kev / tyramine metabolism txoj kev

Cov intermediates tyrosol thiab hydroxytyrosol ntawm dopamine txoj kev / tyramine txoj kev yog lwm qhov tseem ceeb ua ntej rau biosynthesis ntawm Acteoside, thiab lawv tuaj yeem tsim salidroside lossis hydroxytyrosol glucoside, cov khoom ua ntej ntawm Acteoside, los ntawm txoj kev sib txawv. Txoj kev no yog ib txoj hauv kev tseem ceeb hauv txoj kev biosynthesis ntawm Acteoside (Fig. 2b). Tyrosine generates L-dopa raws li kev ua ntawm polyphenol oxidase (PPO) / tyrosine hydroxylase (TH), thiab dopa decarboxylase (DODC) / tyrosine decarboxylase (TyDC) catalyzes L-dopa. -Dopa tsim dopamine, uas tom qab ntawd ua tiav los ntawm thawj amine (muaj tooj liab) oxidase (CuAO) thiab cawv dehydrogenase (ALDH) los tsim hydroxytyrosol [16-18, 24-25]. Hauv lwm ceg, dopamine yog catalyzed los ntawm thawj amine (tooj liab) oxidase (CuAO)/monoamine oxidase (MAO) los tsim 3,4-DHPAA. Vavricka et al[26] siv hydroxyphenylacetaldehyde synthase (DHPAAS) muab tau los ntawm Bombyx mori nrog dual lub luag haujlwm ntawm aldehyde synthase thiab decarboxylase rau catalyze L-dopa rau 3, 4-DHPAA, thiab tom qab ntawd ALDH catalyzed 3, {{14.}}DHPA tsim hydroxytyrosol [24] Txoj kev tyramine tuaj yeem muab tyrosol lossis hydroxytyrosol precursors rau Acteoside biosynthesis txoj kev. Tyrosine yog catalyzed los ntawm TyDC los tsim tyramine, uas yog oxidized rau 4-hydroxyphenylacetaldehyde (4-HPAA) los ntawm CuAO/tyramine oxidase (TYO), thiab 4-HPAA tuaj yeem hloov mus rau {{19 }}hydroxyphenylacetaldehyde reductase los ntawm 4-hydroxyphenylacetaldehyde reductase. (4HPAR)/ALDH raug txo rau tyrosol, thiab tom qab ntawd tyrosol yog catalyzed los ntawm tyrosolhydroxylase (TLH) los tsim hydroxytyrosol [16-17,24,{24}}]. Vim yog lub heterogeneity ntawm TyDC, TYO thiab ALDH, cov kws tshawb fawb tau tso tawm qhov kev xav ntawm lwm txoj kev sib txuas ntawm tyramine, uas yog, tyrosine generates tyramine nyob rau hauv decarboxylation ntawm TyDC, thiab ces tyramine generates dopamine nyob rau hauv qhov kev txiav txim ntawm TH, thiab dopamine yog decarboxylated. los ntawm TH. TYO yog oxidized rau 3,4-DHPAA, uas tom qab ntawd txo mus rau hydroxytyrosol los ntawm ALDH, tab sis TH nyob rau hauv txoj kev putative no tsis tau txheeb xyuas [28]. Qee cov kws tshawb fawb tau xav tias polyphenol oxidase (PPO) tuaj yeem ua rau lub cim ntawm dopamine los ntawm tyramine thiab tiam ntawm hydroxytyrosol los ntawm salidroside los ntawm hydroxylation, tab sis nws txoj haujlwm tsis tau txheeb xyuas [16-18]. Rosmarinic acid yog ib txwm tshwm sim polyphenolic compound, thiab cov kev tshawb fawb tau pom tias tyrosine transaminase (TAT) yog thawj enzyme nyob rau hauv tyrosine-derived txoj kev nyob rau hauv lub synthesis ntawm rosmarinic acid, uas catalyzes lub transamination ntawm tyrosine. 4-Hydroxypyruvate (4-HPPDC) yog tsim, thiab tom qab ntawd 4-HPAA yog tsim los ntawm kev ua ntawm 4-hydroxyphenylpyruvate decarboxylase (HPPADC) [29-30]. Torrens-Spence et al. [31] thawj zaug pom ib pyridoxal phosphate-dependent hydroxyphenylacetaldehyde synthase (4HPAAS), uas tuaj yeem ncaj qha catalyze hloov dua siab tshiab ntawm tyrosine rau 4-HPAA. Ib tug noob encoding ib glycosyltransferase uas catalyzes zus tau tej cov salidroside los ntawm tyrosol. Tsis tas li ntawd, tyrosine tuaj yeem catalyzed los ntawm TYR los tsim hydroxytyrosol [25]. Cov enzymes nyob rau hauv cov enzymatic crossover txoj kev tag nrho enrich ntau haiv neeg ntawm biosynthetic txoj kev ntawm Acteoside. 1.3 Txoj kev hla kev hloov pauv ntawm acyl thiab glycosyl hloov kho ntawm Acteoside

Los ntawm cov qauv analogues ntawm Acteoside, nws tuaj yeem pom tias nws qhov chaw yog tag nrho cov piam thaj, uas yog esterified nrog caffeoyl, thiab C3 txoj hauj lwm yog hloov los ntawm rhamnosyl (Fig. 1a), tab sis nws acylation thiab rhamnosylation Molecular catalytic mechanism kev tshawb fawb tsis tau. qhia. Los ntawm qhov rov qab inference ntawm hydrolysis, kev sim metabolic thiab cov qauv tshuaj ntawm Acteoside, muaj ob lub peev xwm ua tau rau kev sib txuas ntawm Acteoside[19-22]. Ua ntej, caffeoyl-CoA thiab hydroxytyrosol glucoside raug rau kev ua ntawm acyltransferase (quinine hydroxycinnamyl transferase, HCT) los tsim deltoxoside A, uas hloov mus rau rhamnosyltransferase (UDP-rhamnosyltransferase). rhamnose glucosyltransferase, URT) catalyzed ntxiv zus tau tej cov verbasose Lwm txoj hauv kev yog hydroxytyrosol tsim hydroxytyrosol glucoside nyob rau hauv qhov kev txiav txim ntawm glucosyltransferase, thiab hydroxytyroyl glucoside tsim decaffeyl verbasin nyob rau hauv catalysis ntawm URT. Nws condenses nrog caffeoyl-CoA los ua Acteoside (Fig. 2c).

2 Kev tswj hwm zoo ntawm Acteoside synthesis

2.1 Transcriptome mining ntsig txog Acteoside synthesis

Transcriptome yog hais txog cov lej ntawm mRNAs sau los ntawm cov kab mob nyob rau hauv ib qho kev mob ntawm lub cev lossis lub voj voog loj hlob thiab yog ib qho tseem ceeb rau kev kawm cov qauv noob, kev ua haujlwm thiab kev loj hlob. Nws yog ib qho tseem ceeb ntawm kev qhia cov noob thiab cov txheej txheem tseem ceeb ntawm kev tshawb fawb phenotype. Daim ntawv thov ntawm transcriptome thev naus laus zis muab cov cuab yeej zoo rau kev mining ntsig txog enzyme noob hauv biosynthesis ntawm cov khoom nquag ntawm cov nroj tsuag tshuaj. Tseem muaj ntau cov enzymes tseem ceeb hauv txoj kev sib txuas ntawm Acteoside tsis tau daws, yog li mRNA abundance nyob rau hauv cov hlwb nyob rau hauv tej yam physiological tej yam kev mob yuav siv tau los piav qhia cov theem ntawm noob thiab infer lub abundance ntawm cov khoom protein kawg. Los ntawm kev txheeb xyuas cov ntaub ntawv tseem ceeb, cov ntaub ntawv hais txog kev sib txuas ntawm phenylethanoid glycosides tau txais (Table 1), uas tuaj yeem muab tswv yim rau biosynthesis ntawm Acteoside.

Cistanche Tubulosayog ib qho tseem ceeb tshuaj nroj tsuag rauExtract phenylethanoid glycosides, thiab kev kawm ntawm nws cov genome thiab cov ntaub ntawv transcriptome muab ib qho txiaj ntsig zoo rau kev kawm txog txoj hauv kev biosynthetic ntawmActeoside. Sib sib zog nqus transcriptome sequencing ntawm succulent stems ntawmCistanche Tubulosatau nqa tawm siv RNA-seq txoj kev tsom xam rauCistanche Tubulosa, thiab nws cov ntaub ntawv transcriptome tau txais thawj zaug. Los ntawm kev sib piv sib piv thiab kev soj ntsuam phylogenetic, cov enzymes tseem ceeb txuas cov metabolism hauv thawj thiab theem nrab metabolism tau txheeb xyuas. PAL noob, uas tseem yog thawj enzyme tseem ceeb hauv phenylethanoid biosynthesis, thiab 17 enzyme noob koom nrog hauv phenylethanoid biosynthesis hauv rhizomes tau kwv yees thawj zaug [32]. Cov txiaj ntsig ntawm kev tshawb nrhiav qhov zoo sib txawv ntawmCistanche Tubulosanyob rau hauv ntau hom suab puam los ntawm metabolomics thiab transcriptomics pom tau hais tias cov ntsiab lus ntawm phenylethanoid nyob rau hauv saline-alkali av tej zaum yuav siab dua vim lub up-regulation ntawm cov kab lus ntawm PAL, ALDH, aspartate aminotransferase (GOT), thiab lwm yam. [33]. Kev tsom xam ntawm txoj kev hluavtaws ntawm phenethyl glycoside muab lub hauv paus. Scrophulariaceae tsob nroj Rehmannia glutinosa yog nplua nuj nyob rau hauv phenylethanol glycosides. Los ntawm de novo transcriptome sequencing ntawm tuberous cag ntawm Rehmannia glutinosa, thiab muab piv cov KEGG txoj kev thiab txoj kev sim, 19 tus neeg sib tw uas yuav muaj feem xyuam rau biosynthesis ntawm Acteoside yog npaj. noob, uas muab cov kev taw qhia rau kev tsom xam ntawm txoj kev sib txuas ntawm Acteoside. Tom qab ntawd, cov ntaub ntawv tau txheeb xyuas qhov tseem ceeb los ntawm RT-PCR, thiab nws tau pom tias cov ntaub ntawv sau tseg ntawm Rehmannia glutinosa encodes prephenate dehydratase (ADT), aspartate aminotransferase (ASP5), GOT2, 4CL, thawj amine oxidase (AOC3), C4H, HCT. thiab C3H noob [34]. Interestingly, cov ntsiab lus ntawm Acteoside nyob rau hauv cov nplooj ntawm Rehmannia glutinosa yog siab dua li ntawm cov hauv paus hniav, uas tej zaum yuav yog vim hais tias ntawm txawv theem ntawm cov enzymes nyob rau hauv ntau qhov chaw vim hais tias ntawm qhov sib txawv ntawm cov noob nyob rau hauv tej yam kev loj hlob theem.

Tom qab ntawd, plaub qhov sib txawv ntawm Rehmannia glutinosa tau cog qoob loo nrog cov kab txaij radial thiab cov kab txaij tsis-radial hauv cov hauv paus cag, thiab lawv cov ntaub ntawv transcriptome tau txheeb xyuas kom tau txais 223 noob ntsig txog Acteoside synthesis. PAL, C4H, C3H, 4CL, TyDC, PPO, CuAO, ALDH, HCT, UGT thiab lwm yam noob tau txheeb xyuas los ntawm cov ntaub ntawv transcriptome [16]. 231 noob koom nrog hauv biosynthesis ntawm Acteoside tau mined nyob rau hauv transcriptome sequencing ntawm Centantheragrandiflora Benth. 10 cov noob suav nrog PPO thiab HCT. Ntawm lawv, CuAO tau tshaj tawm, ua raws li 4CL, ALDH thiab UGT, thiab qhov tsawg tshaj plaws yog HCT. Genes koom nrog txoj hauv kev phenylalanine thiab tyrosine muaj ntau nyob rau hauv nplooj thiab stems tshaj nyob rau hauv cov hauv paus hniav, tshwj xeeb tshaj yog PAL thiab PPO noob. Thiab piv nrog cov hauv paus hniav, plaub noob PAL, C4H, C3H thiab 4CL nyob rau hauv nplooj thiab stems yog tag nrho cov up-regulated, qhia hais tias phenylalanine-derived txoj kev yog heev nthuav qhia nyob rau hauv cov huab cua qhov chaw [18]. Txawm li cas los xij, cov kev tshawb fawb saum toj no tau kwv yees tias HCT thiab UGT ua rau lub hauv paus enzymatic txoj hauv kev ntawm caffeoyl-CoA thiab hydroxytyrosol glucoside rau Acteoside los ntawm cov kev tshawb fawb transcriptome, tab sis cov haujlwm ntawm cov noob muaj feem xyuam tsis tau txheeb xyuas [16,18]. Inducers hais txog yam uas tuaj yeem ntxias cov nroj tsuag los tsim kev tiv thaiv lossis tiv thaiv tus kheej thiab tiv thaiv, thiab tuaj yeem ua tau sai, tshwj xeeb thiab xaiv ua kom muaj kev qhia ntau yam ntawm cov noob hauv cov nroj tsuag, yog li ua kom cov metabolites theem nrab [35]. Txhawm rau tshuaj xyuas cov elicitor zoo tshaj plaws uas ua rau muaj kev nce ntxiv ntawm Acteoside hauv cov hauv paus plaub hau ntawm Rehmannia glutinosa, cov kev hloov pauv caj ces ntawm Rehmannia glutinosa kho tau los ntawm Agrobacterium rhizogenes tau siv los sib piv cov tshuaj elicitors nrog ntau qhov sib txawv. Nws tuaj yeem tsis tsuas yog txhawb kev loj hlob ntawm cov hauv paus plaub hau, tab sis kuj tseem ua kom cov ntsiab lus ntawm Acteoside. Tom qab ntawd, RNA-Seq thev naus laus zis tau siv los txheeb xyuas cov ntaub ntawv ntawm cov hauv paus plaub hau ntawm Rehmannia glutinosa induced ntawm lub sijhawm sib txawv, kom pom cov noob qhia txawv nyob rau hauv cov xwm txheej sib txawv, thiab thaum kawg los txiav txim siab PAL, C4H, C3H, 4CL, TyDC, PPO, CuAO. , ALDH, HCT, UGT. Isogenic tuaj yeem koom nrog hauv Acteoside biosynthesis. Ntawm lawv, C3H thiab ALDH yog cov noob uas muaj ntau tshaj thiab tsawg tshaj plaws homologous genes, feem [17]. Tsis tas li ntawd, Fatemi et al. [29] soj ntsuam cov txiaj ntsig ntawm ntau qhov sib txawv ntawm cov elicitors xws li methyl jasmonate (MeJA) thiab ntau phab ntsa carbon nanotubes (MWCNTs) ntawm kev sib sau ntawm rosmarinic acid thiab qhia qib ntawm cov noob ntsig txog nws txoj kev biosynthetic. Cov noob ntawm PAL, TAT, 4HPPR thiab rosmarinic acid synthase (RAS) tau raug txheeb xyuas tias yog cov noob tseem ceeb, ntawm cov TAT muaj kev hloov pauv, uas muab lub tswv yim tshiab rau kev tshuaj xyuas ntawm txoj kev sib txuas ntawm Acteoside. Olive (Oleaeuropaea) yog ib qho chaw nplua nuj ntawm bioactive polyphenols, uas tuaj yeem tsim hydroxytyrosol, ib qho tseem ceeb ua ntej ntawm Acteoside. Cov noob muaj feem xyuam rau polyphenol biosynthesis nyob rau hauv txiv ntseej txiv ntoo thiab nplooj raug txheeb xyuas los ntawm tag nrho-ntev transcriptome sequencing. Kaum ob lub tebchaw polyphenol tau suav nrog hauv qhov kev sim, thiab 106 cov ntawv sau tseg ntawm 6 tsev neeg cov noob tau txheeb xyuas thiab hydroxytyrosol synthesis Cov noob muaj feem xyuam rau kev sib txuas ntawm hydroxytyrosol suav nrog PPO, DODC, CuAO, ALDH, thiab PAR [24]. Cov txiaj ntsig ntawm qhov kev tshawb fawb phenethylamine transcriptome saum toj no muab lub hauv paus rau kev tsom xam ntawm txoj kev sib txuas ntawm Acteoside thiab cov mining ntawm kev tswj cov noob thiab cov enzymes tseem ceeb.