天然产物药物生物合成

天然产物药物生物合成的机制提 纲• PKS作用机制及范例• NRPS作用机制及范例• 其他类型天然产物的生物合成提 纲• PKS作用机制及范例• NRPS作用机制及范例• 其他类型天然产物的生物合成聚酮是一种生物聚合物■

单体（合成前体）■

短链羧酸（乙酸或丙二酸）■

单体的顺序聚合■

起始单元+扩展单元■

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

聚酮“核心”骨架的进一步修饰■

利用修饰酶加入新的功能基团DecoratinggroupsMature

polyketide聚酮合酶

Polyketide

Synthase,

PKSKSATACPRed■

聚酮合酶的功能结构域■

酰基转移酶（AT）：选择聚合单体■

酰基载体蛋白（ACP）：使合成单体和合成中间产物共价结合在聚酮合酶上■

酮酯酰合成酶（KS）：连续聚合单体和中间产物■

聚酮合酶其它的功能结构域■

还原活性结构域：使合成的聚酮链完全或部分还原真菌聚酮合酶■真菌聚酮合酶由多结构域组成■

真菌聚酮合酶核心结构域■

所有的真菌聚酮合酶含有AT、KS和ACP核心结构域■

同一套核心结构域用于整个聚酮链的合成KS ATACPReducing

activities■

还原型聚酮合酶（hrPKS）■

含有还原结构域DH、ER和KRKS AT DH ERKR

ACP■

非还原型聚酮合酶（nrPKS）■

含有起始单元酰基转移酶SAT■

决定苯环环化位置的PT结构域■

释放产物的硫酯酶TESAT KS AT PT ACP TEATACPKSTESHSHSHSH最小PKS结构域的组成AT

domain

=

AcyltransferaseAcylCarrier

protein

(ACP)b-ketoacylsynthase(KS)Thioesterase(TE)Next磷酸泛酰巯基乙胺基转移酶磷酸泛酰巯基乙胺基转移酶

(phosphopantetheinyl

transferase,PPTase)功能：可将辅酶A上的4′-磷酸泛酰巯基乙胺转移到载体蛋白(carrier

protein,CP)保守的丝氨酸残基侧链羟基上,使载体蛋白由无活性的脱辅基(apo-)形式转变为活性全蛋白(holo-)形式。ATACPKSTESHSHSHSHAT

domain

=

AcyltransferaseAcylCarrier

protein

(ACP)b-ketoacylsynthase(KS)Thioesterase(TE)辅基:4-磷酸泛酰巯基乙胺(PPT).

比较灵活的基团，可在酶上转移起始和延伸单元。O

OHONHONHSHCH3H3COHH2C O POSerACP最小PKS结构域的组成NextCoAS辅酶

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■

酮酯酰还原酶（KR）：将酮基还原为羟基■

脱水酶（DH）：将羟基还原为双键■

烯醇还原酶（ER）：将双键还原为单键聚酮合酶

Polyketide

Synthase,

PKSI型聚酮合酶(红霉素为例)ProteinModuleDomain大环内酯类抗生素，由菲宾Panay岛上壤中分离的放线菌Streptomyces

erythreus培养液中制成，1952年EliLilly公司首次发表。红霉素

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结构域、模块组织结构和I型聚酮化合物的线形对应关系实例-

OxytetracyclineOxytetracycline

fromStreptomyces

rimosusoxyA oxyCResistanceoxyEMinimal

PKSoxyGoxyIoxyKoxyMKRoxyOCyclaseoxyQoxySotrBoxyBoxyDoxyFoxyHoxyJoxyLoxyNoxyPotrAUnknownoxyRoxyTInitiation

ModuleTailoring02468oxy

Gene

Cluster10 12 141618202224

kBmalonate??oxyAoxyBoxyCoxyDoxyJoxyKoxyNoxyFoxyLoxyQoxyToxySand

othersPKS人物Chaitan

Khosla,

Stanford

University美国工程院院士、斯坦福大学教授，是国际上天然产物生物合成领域最著名的专家之一，在聚酮合酶的研究中取得了世界瞩目的成绩，在乳糜泻病的研究中也有着丰硕成果。现已发表学术论文、申请专利300余篇（项），其中在SCIENCE等国际顶级期刊发表论文10多篇，荣获了多项国际学术大奖，包括美国化学学会的Eli

Lilly奖、2000年美国化学学会的Pure化学奖、2011年的JamesE.Bailey奖。PKS人物Craig

TownsendJohnsHopkins

University真菌PKS生物合成DavidShermanUniversityof

Michigan美国密歇根大学药学院资深教授兼副院长，是国际公认的微生物次级代谢产物生物合成领域的领军人物之一。其研究领域主要包括生物有机化学、分子遗传学、P450酶学、新型药物和天然

产物开发、合成化学和生物合成等方向。提 纲• PKS作用机制及范例• NRPS作用机制及范例• 其他类型天然产物的生物合成核糖体多肽与非核糖体多肽类DOI:

10.1146/annurev.micro.58.030603.123615• 由一系列组件（module）构成• 大多数NRPSs组件数为4-10个，有的高达50个• 每个典型组件由大约1000个氨基酸残基组成• 每个组件负责一个反应循环NRPSs的结构组成NRPS类化合物的结构域基本结构域：腺苷酰化结构域（adenylation，A）缩合结构域（condensation，C）肽酰载体蛋白（peptidyl

carrierprotein，PCP）可选结构域：硫酯酶（thioesterase，TE）差向异构酶（epimerase，E）环化酶（cyclase，CYC）甲基转移酶（methyltransferase，MT）氧化酶（oxi