天然產物藥物生物合成的機制提 綱? PKS作用機制及范例? NRPS作用機制及范例? 其他類型天然產物的生物合成提 綱? PKS作用機制及范例? NRPS作用機制及范例? 其他類型天然產物的生物合成聚酮是一種生物聚合物■

單體（合成前體）■

短鏈羧酸（乙酸或丙二酸）■

單體的順序聚合■

起始單元+擴展單元■

利用聚酮合酶合成聚酮的“核心”骨架StarterExtendersPolyketidecore■

聚酮“核心”骨架的進一步修飾■

利用修飾酶加入新的功能基團DecoratinggroupsMature

polyketide聚酮合酶

Polyketide

Synthase,

PKSKSATACPRed■

聚酮合酶的功能結構域■

酰基轉移酶（AT）：選擇聚合單體■

酰基載體蛋白（ACP）：使合成單體和合成中間產物共價結合在聚酮合酶上■

酮酯酰合成酶（KS）：連續聚合單體和中間產物■

聚酮合酶其它的功能結構域■

還原活性結構域：使合成的聚酮鏈完全或部分還原真菌聚酮合酶■真菌聚酮合酶由多結構域組成■

真菌聚酮合酶核心結構域■

所有的真菌聚酮合酶含有AT、KS和ACP核心結構域■

同一套核心結構域用于整個聚酮鏈的合成KS ATACPReducing

activities■

還原型聚酮合酶（hrPKS）■

含有還原結構域DH、ER和KRKS AT DH ERKR

ACP■

非還原型聚酮合酶（nrPKS）■

含有起始單元酰基轉移酶SAT■

決定苯環環化位置的PT結構域■

釋放產物的硫酯酶TESAT KS AT PT ACP TEATACPKSTESHSHSHSH最小PKS結構域的組成AT

domain

=

AcyltransferaseAcylCarrier

protein

(ACP)b-ketoacylsynthase(KS)Thioesterase(TE)Next磷酸泛酰巰基乙胺基轉移酶磷酸泛酰巰基乙胺基轉移酶

(phosphopantetheinyl

transferase,PPTase)功能：可將輔酶A上的4′-磷酸泛酰巰基乙胺轉移到載體蛋白(carrier

protein,CP)保守的絲氨酸殘基側鏈羥基上,使載體蛋白由無活性的脫輔基(apo-)形式轉變為活性全蛋白(holo-)形式。ATACPKSTESHSHSHSHAT

domain

=

AcyltransferaseAcylCarrier

protein

(ACP)b-ketoacylsynthase(KS)Thioesterase(TE)輔基:4-磷酸泛酰巰基乙胺(PPT).

比較靈活的基團，可在酶上轉移起始和延伸單元。O

OHONHONHSHCH3H3COHH2C O POSerACP最小PKS結構域的組成NextCoAS輔酶

A:

CoenzymA(CoA)O

OHONHONHSHCH3H

C3O POOHO POOHH2COOO P OHOHOHNNNN4-phosphopantetheineAdeninRibo-3’-phosphat=CoenzymAalso

contains

a4-phosphopantetheine

group,

similartothat

foundon

theACP

domain

of

PKSs.

Theterminal

thioester

groupserves

at

theattachment

point

foracetyland

malonyl

units.NH2NextCoASOC CH3ATACPKSTESHSHSHSHLoading

of

astarter

unitStarter

unit(acetyl-CoA)NextATACPKSTESHSHSHSHCoASLoading

of

astarter

unitOC CH3NextCoASHOC CH3ATACPKSTESSHSHLoading

of

astarter

unitSHNextSHCoASHOSH C CH3ATACPKSTESHSHLoading

of

astarter

unitNextSHCoASHOC CH3ATACPKSTESHSSHLoading

of

astarter

unitAstarterunit

has

now

been

loaded

intotheKS

domain

of

thePKSand

weareready

forloading

of

thefirst

extender

unit.NextActivation

of

extender

unitsCoASOC CH3 + CO2Acetyl-CoA

CarboxylaseCoASCOC

H2OC OHTheCO2

originates

from

aHCO3-

bondtobiotin

intheenzymeNextAcetyl-CoAMalonyl-CoASHCoASOC CH3ATACPKSTESHSSHOCC

H2OC OHLoading

of

aextender

unitExtender

unit(malonyl-CoA)NextSHCoAOC CH3ATACPKSTESHSSHOSH CC

H2OC OHLoading

of

aextender

unitNextCoASHOC CH3ATACPKSTESSHC

H2SHOCOC OHSHLoading

of

aextender

unitRasmusJ.N.Frandsen

2007NextSHCoASHOC CH3ATACPKSTESSSHOCC

H2OCO-Ready

forcondensationDecarboxylationof

theextenderunit

(malonyl)provides

theenergy/electron

for

thecondensationNextSHCoASHOC CH3ATACPKSTESSHCC OOOOSHDecarboxylationof

theextenderunit

(malonyl)provides

theenergy/electorne

for

thecodensationCondensationOCC

H2

-NextSHCoASHATACPKSTESHOCC C CH3

H2OSPreparing

fora

secondroundSHNextSHATACPKSTESHOCC C CH3

H2OSCoAOCC

H2OC OHLoading

of

the2nd

extender

unitSHSHNextKSTESHSCoASHATACPC

H2SHOCOC OHSHLoading

of

the2nd

extender

unitOCC C CH3

H2ONextKSTESHCoASHSHATACPSOCCH2SH2nd

condensationOCOOCC C CH3

H2ODecarboxylationNextKSTESHCoASHSHATACPOCC C CH3

H2OOCCH2SHRelease

fromthe

enzymeSAt

this

stagethe

enzymefaces

a

choice,whether

to

continue

with

additionalroundsof

condensations

ortorelease

thepolyketide

chainfrom

the

enzyme.NextThenumber

of

condensation

rounds(iterations)that

the

individual

PKSsperform

isatpresent

not

predictable.

Onehypothesis

isthat

the

size(volume)

of

theactivesitein

the

KS

domaincould

bethedeciding

factor

for

total

number

ofiterations

possible.KSTESHCoASHSHATACPOCC C CH3

H2OCH2OSH CSHRelease

fromthe

enzymeNextSKSTECoASHSHATACPOCC C CH3

H2OOCCH2SHRelease

fromthe

enzymeSHNextSKSTECoASHSHATACPSHRelease

fromthe

enzymeSHOCC C CH3

H2OOCCH2NextKSTECoASHSHATACPSHRelease

fromthe

enzymeSHSHOCC C CH3

H2OOCCH2HONextKSTECoASHSHATACPSHRelease

fromthe

enzymeSHSHOCC

H2OC CH3OCCH2Starter

unit1st

extender

unit2nd

extender

unitHONextRelease

fromthe

enzymeOCH2 COC C CH3

H2OCHONote

that

the

formed

polyketide

chain

has

polarity.With

amethyl(-CH3)

group

atthe

”oldest”

end

and

acarboxyl

(-COOH)

group

atthe

”newest”

end.NextWhere

does

the

diversity

originate

from?In

addition

to

the

four

catalytic

domains

(AT,ACP,

KS

and

TE)

used

by

the

minimal

PKS.Other

domains

canalso

participate

in

the

biosynthesis:b-ketoacyl

reductase

(KR)Dehydratase

(DH)Enoyl

reductase

(ER)Methyltransferase

(MET)Cyclases

(Cyc)

–

foldthe

polyketide

chain

into

anaromatic

ormacrocyclic

compound+

alternative

extenderunits

differentfrommalonyl-CoACH2COCHH2COHCHH2COHCHHCCHHCCH2H2CCHH2COHCHH2CO CH3ENDOR1R2 R3OSOH OR1R2 R3SR1R2 R3OSR1R2 R3OSDHERKRACPACPACPACP■

酮酯酰還原酶（KR）：將酮基還原為羥基■

脫水酶（DH）：將羥基還原為雙鍵■

烯醇還原酶（ER）：將雙鍵還原為單鍵聚酮合酶

Polyketide

Synthase,

PKSI型聚酮合酶(紅霉素為例)ProteinModuleDomain大環內酯類抗生素，由菲賓Panay島上壤中分離的放線菌Streptomyces

erythreus培養液中制成，1952年EliLilly公司首次發表。紅霉素

PKS

(DEBS)S SSS SSSOO

OHO

OH

OHOO

OH

OHOO

OH

OHO

OHO

OH

OH1

O

OHOHAT

ACP

KS

AT

KR

ACPKS

AT

KR

ACPKS

AT

KR

o

ACP

KS

AT

DH

ER

KR

ACPE13O115

OModule

3Module

4Module

5Module

615O

OH

OHDEBS

1DEBS

2DEBS

3Module

2OOHOH9OH9Load Module

1DEBPeterLeadlay

etal.(Cambridge)1990Leonard

Katzetal.(AbbottLabs)

1991KS

AT

KR

ACP

KS

AT

KR

ACP

TAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPSH SH SHKS

AT

ACP

KS

AT

DH

ER

KR

ACPSH SHKS

AT

KR

ACP

KS

AT

KR

ACP

TESH SHmodule

1

module

2

module

6

module

5

module

4module

3LoadS S S S S S SO O O O O O OCOO- COO- COO- COO- COO- COO-AT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPS SO OCOO- COO-KS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OCOO- COO-SOS SCOO-OCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DHER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OSOCOO- COO-COO- COO- COO-S SCOO-OOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DHER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OSOCOO- COO-COO- COO- COO-S SCOO-OOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPS SKS

AT

ACP

KS

AT

DHER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OCO2OOCOO- COO-COO- COO- COO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

ATKR

ACP

KS

AT

KR

ACPS SKS

AT

ACP

KS

AT

DH

ER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OOOHCOO- COO-COO- COO- COO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPS SKS

AT

ACP

KS

AT

DHER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OOOHCOO- COO-COO- COO- COO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DHER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OSOHCO2O COO- COO-COO- COO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DHER

KR

ACPS SO OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OSOHOHCOO- COO-COO- COO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DHER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OSS SOOHOHCOO-OCOO- COO- COO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPS SKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OCOO- COO-COO-OOOHOHCO2Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPCOO- COO-COO-OKS

AT

ACP

KS

AT

DH

ER

KR

ACPS SKS

AT

KR

ACP

KS

AT

KR

ACP

TES SO OOOHOHOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TES SOOHOHCO2OSOCOO-OCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TES SOOHOHOHSOCOO-OCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

ATDHER

KR

ACPOOHOHOCOO-OKS

AT

KR

ACP

KS

AT

KR

ACP

TES SSCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TES SCOO-OCOO-OOOHOHSOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TES SCOO-OCOO-OOOHOHSOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOHOHCO2SOOSCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

ATKR

ACP

KS

AT

KR

ACP

TEOOHOHSOOHSCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOHOHSOOHSCOO-Omodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOHOHCO2SOOOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

ATKR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOHOHSOOOmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOOSmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOOOHOHOHmodule

1

module

2

module

6

module

5

module

4module

3LoadAT

ACP

KS

AT

KR

ACP

KS

AT

KR

ACPKS

AT

ACP

KS

AT

DH

ER

KR

ACPKS

AT

KR

ACP

KS

AT

KR

ACP

TEOOOOHOHOHOHHOmodule

1

module

2

module

6

module

5

module

4module

3LoadDesosamineCladinoseSSSSSSSOOOHOOHOHOOOHOHOOO

HO

HOO

HOO

HO

HOO

HO

HAT

ACPKS

AT

KR

ACPKS

AT

KR

ACPKS

AT

KRo

ACPKS

AT

DH

ER

KR

ACPKS

ATKR

ACPKS

AT

KR

ACP

TE13O115

O

Module

3

Module

4

Module

5

Module

615OO

HO

HDEBS1DEBS2DEBS3Module

2O9O

HO

HOH91Load Module

1O113O15

OO

HO

HOH913O115

OO9O

HOHOPKS結構域、模塊組織結構和I型聚酮化合物的線形對應關系實例-

OxytetracyclineOxytetracycline

fromStreptomyces

rimosusoxyA oxyCResistanceoxyEMinimal

PKSoxyGoxyIoxyKoxyMKRoxyOCyclaseoxyQoxySotrBoxyBoxyDoxyFoxyHoxyJoxyLoxyNoxyPotrAUnknownoxyRoxyTInitiation

ModuleTailoring02468oxy

Gene

Cluster10 12 141618202224

kBmalonate??oxyAoxyBoxyCoxyDoxyJoxyKoxyNoxyFoxyLoxyQoxyToxySand

othersPKS人物Chaitan

Khosla,

Stanford

University美國工程院院士、斯坦福大學教授，是國際上天然產物生物合成領域最著名的專家之一，在聚酮合酶的研究中取得了世界矚目的成績，在乳糜瀉病的研究中也有著豐碩成果。現已發表學術論文、申請專利300余篇（項），其中在SCIENCE等國際頂級期刊發表論文10多篇，榮獲了多項國際學術大獎，包括美國化學學會的Eli

Lilly獎、2000年美國化學學會的Pure化學獎、2011年的JamesE.Bailey獎。PKS人物Craig

TownsendJohnsHopkins

University真菌PKS生物合成DavidShermanUniversityof

Michigan美國密歇根大學藥學院資深教授兼副院長，是國際公認的微生物次級代謝產物生物合成領域的領軍人物之一。其研究領域主要包括生物有機化學、分子遺傳學、P450酶學、新型藥物和天然

產物開發、合成化學和生物合成等方向。提 綱? PKS作用機制及范例? NRPS作用機制及范例? 其他類型天然產物的生物合成核糖體多肽與非核糖體多肽類DOI:

10.1146/annurev.micro.58.030603.123615? 由一系列組件（module）構成? 大多數NRPSs組件數為4-10個，有的高達50個? 每個典型組件由大約1000個氨基酸殘基組成? 每個組件負責一個反應循環NRPSs的結構組成NRPS類化合物的結構域基本結構域：腺苷酰化結構域（adenylation，A）縮合結構域（condensation，C）肽酰載體蛋白（peptidyl

carrierprotein，PCP）可選結構域：硫酯酶（thioesterase，TE）差向異構酶（epimerase，E）環化酶（cyclase，CYC）甲基轉移酶（methyltransferase，MT）氧化酶（oxi