Acetate Pathway PDF

Acetate Pathway Primary Metabolism - Overview Primary metabolites Primary metabolism Secondary metabolites CO2 + H2O PHOTOSYNTHESIS -그 1) ’light reactions1: hv ATP and NADPH 2) [dark reactions1: CO? -> sugars fCa/v/n cyc/e) SHI KI MATE METABOLITES cinnamic acid derivatives □romat/c compound...

Acetate Pathway Primary Metabolism - Overview Primary metabolites Primary metabolism Secondary metabolites CO2 + H2O PHOTOSYNTHESIS -그 1) ’light reactions1: hv ATP and NADPH 2) [dark reactions1: CO? -> sugars fCa/v/n cyc/e) SHI KI MATE METABOLITES cinnamic acid derivatives □romat/c compounds lignans, flavinoids ALKALOIDS penicillins cephalosporins cyclic peptides FATTY ACIDS & POLYKETIDES prostaglandins polyacetylenes □ron?3t/c compounds, polyphenols macrolides ISOPRENOIDS terpenoids steroids carotenoids 2 Acetate Pathway Biosynthesis of Malonyl Coenzyme A Malonyl coenzyme A is the key ‘extender unit’ for the biosynthesis of fatty acids (<& polyketides): - is formed by the carboxylation of acetyl coenzyme A mediated by a biotin-dependent enzyme - this is the first committed step of fatty acid/polyketide biosynthesis (& is a rate controlling step) Secondary metabolism FATTY ACIDS & POLYKETIDES prostaglandins po/yacety/enes aromatic compounds, polyphenols macrolides mevalonate (MVA) ISOPRENOIDS terpenoids steroids carotenoids Acetate Pathway (Fatty Acid) CO2H acyl carrier protein (ACP) 1 CH3CO-SCOA acetyl-CoA CO2H 1 CH2co-SCoA malonyl-CoA 1 1 1 CHoCO-S-ACP malonyl-ACP I Claisen reaction H OH RCH2c0 —S —Enz acyl-enzyme thioester NADPH RCHf^CH2co-S-ACP RCH2coeH2co-S-ACP each turn of the cycle extends the chain length of the acyl group by two carbons P-keto acyl-ACP p-hydroxy acyl-ACP stereospecific reduction of carbonyl £2 eliinination ofH2O RCH2 스〜f,CO —S—ACP aJ3-unsaturated acyl-ACP NADPH reduction of double bond RCH2cH2cH2co-S-ACP fatty acyl-ACP HSCoA RCH2cH2cH2co -SCoA fatty acyl-CoA RCH2cH2cH2co2H fatty acid 4 Acetate Pathway (Fatty Acid) SCoA HOOC SCoA malonyl CoA C2 unit Acetyl CoA SE SE Reduction SE COOH //느COOH 5 Acetate Pathway(Aromatic Polyketides) Acetate Pathway (Aromatic Polyketides) 7 8 Senna - ■ ■ Cassia angustifolia, Cassia senna （ 차풀） a stimulant laxative and acts on the wall of the large intestine, increasing peristaltic movement 9 Flavonoids and Stilbenes (shikimate pathway + acetate pathway) C6C3 from shikimate pathway (a stilbene) Resveratrol? 3,5,4'-trihydroxystilbene and its glucoside ■ 포도껍질, 포도씨, 적포도주, 땅콩 등에 함유 ■ 식물이 적대적인 환경에 처했을때 만들어 내는 파이토알 렉신(phytoallexin) ■ Antioxidant, antimutagenic, antiinflammatory, anti­ tumor promoting, cancer preventive effect ■ ■ Esp. rich in wine 으 moderate red wine consumption may reduce the risk of cardiovascular disease 11 History of Resveratrol 지b-------------------------------------(1976) 포도의 껍질과 씨앗에 존재하는 resveratrol 발견 今포도 및 와인의 품종에 따라 매우 다양한 농도로 함 유 되어 있음 ■ • ■ Resveratrol found in Red Wine. French Paradox : 프랑스인들이 고지방 고단백 식사를 즐겨하는 반면 심혈관 질환이 낮은것이 와인 음용과 관련이 있다는 것을 발견 12 Tetracyclines R1 tetracycline chlortetracycline oxytetracycline demeclocycline H H OH H R2 6a Me Me Me H methacycline doxycycline minocycline OH OH H Me H lymecycline H R3 R4 6P OH OH OH OH H Cl H H =CH2 Me H H OH natural H H NMe2 H H H H 心 semi’ synthetic H 스yc02H NH2 . Streptomyces species ■ Broad spectrum antibiotics ■ inhibition of protein synthesis (ribosome) ■ Metal ion chelator ■ Tetracycline Biosynthesis oxidation of 1,4-qumol To quinone hydroxylalion at activated centre para to phenol 6-methy 1 prele Ira m i de tautomerism To keto form, followed by hydration of double bond chlorination at activated position para to phenol reduction 으/ double bond of oc,p-unsaturated ketone; eno! for. then favoured due to conjugation NMc hydroxy I a l ion; consider the tautomeric Tetracyclines R1 2 R2 6a MX Mk tetracycline chlortetracycline oxytetracycline demeclocycline 므 methacycline doxycycline minocycline OH OH H lymecycline Mk H H R3 R74 oP H H o H clH o H H o H =CH2 Me H HH Me OH H H natural H H H H NMe2 H H H H 오 semisvnthetic cH XX 、〜厂 느COiH 2 느^、才 NH2 14 Macrolides: 14-16 membered lactone ring HO、 OH T X OMeH zearalenone tacrolimus (FK506) o MeCKr rifamycin B 15 Macrolides Biosynthesis: Polyketide Synthase (PKS) —— Fate of carbonyls: O ； SEnz 、、、애 CO2H SCoA /?o reduction; cyclization to < aromatic ring L ^SCoA O reduction dehydration reduction no reduction PKS see Figure 3.60 reduction dehydration aldol reaction, aromatization, lactone formation SEnz 11 reduction dehydration reduction reduction (ketoreductase) OH —CH-CHn — dehydration t (dehydratase) 스、 y、느人으o 、스、스、 zearalenone reduction enzyme-bound partially-reduced intermediate reduction T (enoylreductase) —CH2-CH2- The Polyketide Pathway Polyketides are also sometimes known as acetogenins acetyl CoA is also the starting point for the biosynthesis of polyketide secondary metabolites these metabolites are topologically very different to the fatty acid metabolites b니t are synthesised in a very similar fashion. The difference is that d니ring the iterative cycle of chain extension the p~k은to group /s generally not completely reduced out. This gives rise to luige structural diversity based aro니nd a 1,3-oxygenation pattern & cyclisation to give aromatic compounds the polyketide pathway NB. unlike fatty acids, polyketides are NOT biosynthesised by humans - only microorganisms (bacteria) & fungi Macrolides Biosynthesis: Polyketide Synthase (PKS) SEnz SEnz SEnz SEnz SEnz SEnz SEnz SEnz SEnz HO- HO- HO zearalenone 17 Polyketides the structural variety of polyketide secondary metabolites is very wide: - NB. starter units marked in red; extender units in bold black; post oligomerisation appended groups in blue 6-methylsa l icy lie acid (antibiotic) citrinin (kidney toxin 'yellow rice disease') Griseofulvin (treatment for nng worm infections) actinorhodin (antibiotic) POLYKETIDES O aflatoxin B1 (mycotoxic carcingen) NB. a mixed polypropionate/acetate 6-deoxyerythronolide B NB. a polypropionate (antibiotic) (anti-cholesterol) Avermectin Methyl malonyl-CoA CO,H SCoA SCoA SCoA + 7x o Malonyl-CoA CO,H spi ro-ket시 )으 H 노^、!JO McO L-oleandrose avermectin B|a aglycone L-olcandrosc avcrnicclin B2a avermectin B COH SCoA o L-Val 「用 avermcciin ivermectin 18 Avermectin - 16-membered macrolides - Anthelmintic （ 구충） , insecticidal - low toxicity to animal and human - Streptomyces avermectilis 19 2015 노벨생리의학상 수상자 오무라 사토시（80） 일본 기타사토（北里）대 교수 - 오무라 특별영예교수는 늦게 학자의 길에 들어섰지만 '사람들에게 도움이 되는 것을 생각하라'는 할머니의 가르침을 가슴에 새기고 끈 기있게 연구에 매달렸다. - 그를 비롯한 연구팀원은 늘 작은 비닐봉지를 지니고 다니며 여기 저기서 흙을 채취해 미생물을 연구했고 이런 과정에서 시즈오카현 의 한 골프장 인근에서 가져온 토양에서 이버멕틴을 만드는 균이 발 견됐다. - 연구 성과로 만들어진 약품은 매년 수억 명에게 투여돼 실명의 위 험을 줄이고 있다. - 200억 엔（약 1천934억원） 이상의 특허료 수입을 기타사토（北里） 연구소에 안겨주고 더 안정적인 연구 기반을 조성하는 계기가 됐다. 20 Polyene Antifungals nystatin At Polyene Antifungals 거^-- -----▪Macrocyclic lactones: 26-38 membered rings ▪ conjugated E double bonds ▪ antifungal, no antibacterial ▪ binding to sterol in the eukaryotic cell membrane ▪ polyene bind about 10-fold more strongly to ergosterol (fungi) than cholesterol (mammal) ▪ UV今destruction 22 Tacrolimus (FK-506) ，-H ■ Streptomyces tsukubaensis ■ macrolide immunosuppressant (FK506) ■ used in liver and kidney transplant surgery ■ inhibits T-cell activation ■ binds first to a receptor protein, which then inhibits a phosphatase enzy me called calcineurin; prevent appropriate gene transcription and subseque nt T-cell activation Tacrolimus (FK-506) ▪ Discovered in 1984 from the fermentation broth of a Japanese soil sample that contained the bacteria Streptomyces tsukubaensis [ Streptomyces tsukubaensis No.9993 ] 25 __ Tacrolimus (FK-506) ▪ Tacrolimus (also FK506 or Fujimycin) Prof. Jon Clardy (Harvard Medical School) ▪ Macrolide antibiotic ▪ First material of Chemical biology: FKBP ▪ Immunosuppressive drug ▪ First approved by FDA in 1994 for use in liver transplant surgery 23 FK-506 Biosynthesis CO,H CO2H SCoA 十 5 x SCoA 0소U즈人r O ----------------------cyclohexanecarbonyl-CoA (from shikimic acid) CO,H CO,H Methyl malonyl-CoA SCoA O propyl inalonyl-Co A 〈5 pipecolic acid HO、 HO、 MeO MeO SEnz I PKS H M 。 OMe tacrolimus (FK506) 26 Tacrolimus (FK-506) ▪ similar compounds 4--------- tacrolimus :- Ciclosporin (cyclosporin) Pimecrolimus Geldanamycins Biosynthesis Loading Mod 니 e 1 Mod니e 2 Mod니e 3 Module 4 Mod니e 5 Mod니e 6 Mod니e7 CoLACP KS AT DH ERKRACP KS AT DH ERKRACP KS AT KR ACP KS AT DH KRACP KS AT KR ACP KS AT DH ER KRACP KS AT DH KRACP i. ? 우 ? ? ； c S H3CO i'Y H3COl'1' H3C0 N' .〉=° WJH H3coiH H3COb' H 3co "L, 3-amino-5-hydroxy benzoic acid (AHBA) > H 3co H3coiM NH2 H3COII' H3C— NH2 HO ACP: acyl carrier protein KS: beta-ketoacyl synthase AT: acyltransferase KR: beta-ketoacyl reductase H3C— Amide synthase Post-PKS modification Geldanamycin Progeldanamycin Antibiotics derived from AHBA Rifamycin B Ansamitocin P-3 브 3-amino-5-hydroxy benzoic acid (AHBA) Geldanamycin Mitomycin C Epothilone = Epoxide + thiazole + ketone Sixteen membered macrolides ■ Epothilone A, R = H ■ Epothilone B, R = CH3 ■ Epothilone A (R = H) E pothilone B (R = CH3) Epothilone C (R = H) E pothilone D (R = CH3) EpothilonesE (R = H) E pothilone F (R = CH3) Sorangium cellulosum - gram-negative myxobacterium - potency determined (1995 ) - first identified as antifungal activity - activity in a tubulin polymerization assay - studied as potential antitumor agents - epothilones A to F have been identified Disadvantage of S. cellulosum - long doubling time (16 h) - difficult to engineer (DNA into the bacterium ) - - limited number of molecular tools and markers that have been developed 28 Structural similarities 1. More active than Taxol (in vitro), competitive inhibitors of taxol binding 2. Comparatively simple in structure 3. Active against Taxol resistant cancer cell lines 4. Easier to derivatize than Taxol 5. More water soluble than Taxol 29 Biosynthesis of epothilones The So Ce90 genome sequenced: Nine modules of polyketide synthase (PKS) + One nonribosomal peptide synthetase (NRPS) + Cytochrome P450 monooxygenase (epoxidation) Module 2 M0dde 3 Module 4 wChem. Biol. 2000, 】 7, 97 Moduh 9 30 Biosynthesis of epothilones -- Post Assembly Line Epoxidation N 丄、必、』 --- > Cyclorelease HSEnz TE domain TE domain Epothilone D P450 Monooxygenase o OH o Epothilone C Epothilone A Epoth ilone B 31 Epothilone A & β-tubulin Fig. 3. Hydrogen bonding (violet) around EpoA in p-TB. Oxygens from C1 to C7 engage in network H bonds with M-loop residues. The thiazole is anchored by His227. Disruption of primary or secondary hydrogen bonds would occur upon mutation of Ala231, Thr274. Arg282, or Gin292 to other residues as observed in epothilone-resistant cells. Pro­ tein secondary structure for he­ lices is shown in red, sheets in blue, and loops in yellow. The protein side chains are colored by atom type: white, C; red, 0; and blue, N. The EpoA ligand is colored by atom type: orange. C; red, O： blue, N： and yellow, S. Microtubules - a fundamental role in diverse cellular functions such as cell division, growth, and motility - If this function is disrupted, a cell cycle arrest at the G2/M phase occurs, which in turn results in apoptosis. Science vol. 305 p. 866-869 (2004) Products & research of epothilones ■ ■ Epothilone (NSC 703147) EpoB quite toxic in an in vivo mouse setting. toxic epoxide moiety of epothilone B was removed ■ 12,13-desoxyepothilone B (dEpoB, NSC 703147), which had been shown to be much less toxic than epothilone B. ■ KOSAN Biosciences obtained the license. 32