Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

MC LC
M U ................................................................................................................ 3 CHNG 1: GII THIU CHUNG V ACID AMIN............................................ 4 1.1. 1.2. Cng thc cu to ....................................................................................... 4 Phn loi .................................................................................................... 6

1.2.1 Da vo cu to ha hc chia thnh mt s nhm ln sau: ....................... 6 1.2.2. Da vo cu trc, acid amin c chia ra lm 3 nhm ln :.................... 7 1.3. Cc con ng tng hp acid amin ................................................................ 8

1.3.1.Sinh tng hp acid amin bng phn ng amin ha trc tip ......................... 8 1.3.2.Sinh tng hp acid amin bng phn ng chuyn amin ............................... 10 CHNG 2: CC CON NG SINH TNG HP ACID AMIN VI SINH VT .................................................................................................................... 13 2.1. T ketoacid ........................................................................................... 14 2.1.1. T ketoglutarate .............................................................................. 14 2.1.1.1. Glutamate / Glutamine ....................................................................... 14 2.1.1.2. Proline ............................................................................................... 17 2.1.1.3. Arginine ............................................................................................. 20 2.1.2. T oxaloacetate ..................................................................................... 21 2.1.2.1. Aspatate/ Asparagine ......................................................................... 21 2.1.2.2. Methionine ......................................................................................... 24 2.1.2.3. Threonine ........................................................................................... 25 2.1.2.4. Lysine ................................................................................................. 25 2.1.3. T pyruvat ............................................................................................ 29 2.1.3.1. Alanine ............................................................................................... 29 2.1.3.2. Valine v Leusine ............................................................................... 29 2.1.3.4. Isoleusine ........................................................................................... 32 2.2. T phosphoglycerate ................................................................................... 33 2.2.1.Serine ..................................................................................................... 33 2.2.2. Glycine ................................................................................................. 34
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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.2.3. Cystein .................................................................................................. 42 2.3. Cc amin thm ............................................................................................ 44 2.3.1. Tng hp ra tin cht trung gian Chosrimate. ........................................ 47 2.3.2. Sinh tng hp Phenylalanine v Tyrosine .............................................. 49 2.3.3. Sinh tng hp Tryptophan ..................................................................... 50 2.4. Histidine ...................................................................................................... 52 CHNG 3: KT LUN ................................................................................... 54 TI LIU THAM KHO ................................................................................... 55

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

M U
S sng l mt qu trnh trao i cht lin tc. Qu trnh trao i cht l qu trnh i mi thnh phn c th bng cch thu nhn cc cht dinh dng t thc n, nc ung v c th thc hin mt qu trnh sinh l, sinh ha phc tp sau o thi cc cht cn b ra ngoi. Thng qua qu trnh ny, c th ln ln, sinh sn duy tr ni ging cui cng l gi ci ri cht i. l qu trnh sinh dng bnh thng ca c th sng, c rt nhiu cc cht tham gia vo qu trnh ny trong mt thnh phn khng th thiu l protein. C th ni protein l thnh phn rt quan trng ca c th, protein c mt trong nhn, nguyn sinh cht, mng t bo, huyt tng, dch gian bo. Acid amin l cu t c bn ca protein, hay ni mt cch khc, acid amin l nhng vin gch xy nn cc ta lu i mun hnh nghn v ca phn t protein. V vy vai tr acid amin rt quan trng, lun l i tng con ngi nghin cu tng hp m c bit l cc acid amin khng thay th.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

CHNG 1: GII THIU CHUNG V ACID AMIN

1.1. Cng thc cu to

Acid amin l nhng hp cht hu c mch thng hoc mch vng trong phn t c cha t nht mt nhm amin (-NH2) v mt nhm cacboxyl (-COOH). Hin nay, c khong hn 100 loi acid amin c con ngi tm thy trong t nhin hay tng hp c. Tuy nhin trong s ny ch c khong 20 loi acid amin l c c th s dng tng hp protein. Trong gm acid amin thit yu v acid amin khng thit yu. Con ngi v nhiu loi ng vt khc ch c th t tng hp c 10 loi acid amin, chng c gi l cc acid amin khng thit yu (Non-essential), 10 acid amin cn li c th khng t tng hp c m phi hp th t thc vt v ng vt qua con ng thc n, chng c gi l nhng acid amin thit yu (Essential). Khi thiu, thm ch ch mt trong cc acid amin cn thit c th lm cho protein c tng hp t hn protein b phn gii, kt qu l cn bng nit m. Cc acid amin cn thit i vi c th cn ty thuc vo iu kin ring bit vo loi ng vt, la tuiTheo nhiu ti liu c tm acid amin cn thit cho ngi ln valine, leucine, isoleucine, methionine, treonine, phenylalanine, triptophan v lysine. i vi tr em c thm hai acid amin cn thit : arginine v histidine. Hm lng cc acid amin khng thay th v t l gia chng trong phn t protein l mt tiu chun quan trng nh gi cht lng protein. Cc acid amin thng gp trong cc protein trong t nhin l nhng L acid amin c nhm amin nh vo nguyn t cacbon ng cnh nhm cacboxyl.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Bng 1.1: Cc acid amin thit yu v khng thit yu Nonessential Essential

Alanine

Arginine*

Asparagine

Histidine

Aspartate

Isoleucine

Cysteine

Leucine

Glutamate

Lysine

Glutamine

Methionine*

Glycine

Phenylalanine*

Proline

Threonine

Serine

Tyrptophane

Tyrosine

Valine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

1.2. Phn loi

1.2.1 Da vo cu to ha hc chia thnh mt s nhm ln sau: a. Nonpolar aminoacid * Glycine, Alanine, Valine, Leucine, Isoleucine * Phenylalanine, Tryptophane, Methionine * Proline, Cysteine b. Polar aminoacid (neutral) * Asparagine, Glutamine, Serine * Threonine, Tyrosine c. Basic amino acids * Histidine, Lysine, Arginine d. Acidic amino acids * Aspartate Glutamate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Hnh 1.1: Phn loi acid amin da vo cu to ha hc 1.2.2. Da vo cu trc, acid amin c chia ra lm 3 nhm ln : Acid amin mch thng (glycine, alanine, leucine, iosleucine, valine, threonine, aspartate, asparagine, glutamate, glutamine, lysine, arginine, methionine, serine, cysteine)

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Amino acid vng thm (phenylalanine, tyrosine) Amino acid d vng (tryptophan, histidine, proline) C th tng hp cc acid amin bng phng php ha hc, phng php ln men hoc bng phng php kt hp ha hc v vi sinh vt. Tuy nhin phng php ha hc thng l t v khng nh hng c: cc acid amin tng hp c cha c hai dng D v L. Phng php kinh t v trin vng hn c l sinh tng hp hay l tng hp bng phng php ln men.

1.3. Cc con ng tng hp acid amin

1.3.1.Sinh tng hp acid amin bng phn ng amin ha trc tip S amin ha trc tip cc acid hu c cha no l mt phn ng t gp v ch yu ch c thc vt v vi khun. Cc cht tin thn amin ha nh acid fumaric, acid piruvic, acid oxaloacetic l nhng sn phm trung gian vn c to nn trong chu trnh Krebs. Acid fumaric, di tc dng ca enzyme aspartat amoniac liza to nn acid aspartic: aspartatamoniacliase + NH3

Phn ng ny l phn ng thun nghch. Acid aspartic c th tng hp bng cch chuyn ha gia acid oxalacetic v acid glutamic. S amin ha cc cetoacid cng l mt ng hng ch yu ca vic tn to acid amin. Phng trnh phn ng tng qut c th c biu din nh sau:

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Phn ng ra qua hai giai on: kt hp amoniac vo cetoacid to ra acid imin v kh acid imin thnh acid amin. Chng hn:

Hoc nh:

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

V nguyn tc, s amin ha c th tin hnh i vi bt k mt cetoacid no. Tuy nhin hot tnh ca tt c cc dehydrogenase t nhin ca cc acid amin, tr cc glutamatdehydrogenase v alanindehydrogenase u khng ng k. V th s tng hp cc acid amin bng amin ha cc cetoacid l khng c ngha thc t. Cc acid amin: aspartic, alanine, glutamic l nhng acid amin s cp. S sinh tng hp bng phn ng amin ha trc tip ny gi l sinh tng hp s cp cc acid amin. 1.3.2.Sinh tng hp acid amin bng phn ng chuyn amin Phn ng chuyn amin cng l mt ng hng sinh tng hp cc acid amin vi sinh vt. Phn ng chuyn amin c th biu din di dng tng qut nh sau:

R1 CH COOH NH2

R2 C COOH O

R1 C COOH O

R2 CH COOH NH2

acidamin 1

cetoacid 2

cetoacid 1

acidamin 2

Phn ng chuyn amin c xc tc bi enzyme aminotransferase. Aminotransferase l enzyme hai cu t c nhm ngoi l phosphopiridoxal (este phosphorid ca vitamin B6). C ch phn ng nh sau: u tin nhm ngoi ca enzyme s kt hp vi nhm amin ca acid amin 1 nhm aldehid ca phosphopiridoxal.
CH = O HO H3C CH2O N enzyme
P

CH = N CH H2N CH R1 COOH Protein c cht

R1 COOH
P

2.

- H2O

HO H3C N

CH2O

Protein

phc enzyme c cht

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Tip phc hp c cht enzyme chu s h bin:

CH = N CH HO H3C N CH2O

R1 COOH
P

CH2 N = C HO H3C N CH2O

R1 COOH
P

Protein

Protein

Phc hp b thy phn gii phng ra cetoacid 1 v enzyme:

CH2 NH2 HO + H2O COOH R1 C = O H3C N Protein + H3C N CH2O
P

CH2 N = C HO CH2O

R1 COOH
P

Protein

cetoacid 1 n lt enzyme li tng tc vi cetoacid 2:

R2 CH2 NH2 HO H3C N CH2O
P

R2 + Protein C=O COOH - H2O H2O

CH2 N = C COOH HO H3C N CH2O

P

Protein

enzym e
R2 CH2 N = C COOH HO H3C N CH2O
P

cetoacid 2

phc c cht enzyme

R2 CH = N CH COOH HO H3C N

h bin

CH2O

Protein

Protein

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Cui cng acid amin 2 v enzym trng thi u c hnh thnh do kt qu thy phn phc hp c cht enzym va to thnh:
R2 CH = N CH COOH HO H3C N CH2O
P

CH = O HO H2O H3C CH2O N

P

R2 C NH2 + Protein COOH

Protein

enzyme trng thi ban u

acidamin 2

Cn ch rng phn ng chuyn amin khi khng c enzym xc tc, cng s tin hnh c trong iu kin nhit 1000C, pH t 3,8, c ion kim loi v piridoxal tham gia.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

CHNG 2: CC CON NG SINH TNG HP ACID AMIN VI SINH VT

S CHUYN HA CC ACID AMIN VI SINH VT

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.1. T ketoacid
2.1.1. T ketoglutarate 2.1.1.1. Glutamate / Glutamine
-O

H C H2 C H2 C C NH3+ COO -

Glutamate c to ra do qu trnh chuyn amin, qu trnh m nhm NH2 ca mt acid amin c chuyn ti - ketoacid vi s xc tc ca transaminase. Mt loi - ketoacid kh ph bin trong t bo vi sinh vt l - ketoglutarate, hp cht trung gian trong vic cung cp nng lng hoc cht nn ca cc qu trnh chuyn ho khc nhau. Glutamate c tng hp kh d dng qua phn ng thay th gc phosphate bi nhm amin vo tin cht - ketoglutarate vi s tham gia xc tc ca glutarate dehydrogenase. Ngoi ra glutamate cn l tin cht tng hp mt s acid amin khc nhau nh glutamine, proline v arginine. Glutamic acid l dng proton ca glutamate. Enzyme glutamatdehydrogenase xc tc cho s tn to ra acid glutamic t ketoglutarate v ammoniac khi c s tham gia ca coenzyme NAD.H2 hoc NADP.H2. Enzyme ny rt ph bin trong h thng sng. NADP.H2 (dng kh) thng c s dng trong cc trng hp khi m glutamatdehydrogenase c chc nng nh mt tc nhn sinh tng hp, cn NAD.H2 (dng kh) c s dng trong cc trng hp khi enzyme ny hon thnh chc nng d ho.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

COO C CH2 CH2 COO alpha-ketoglutarate O + NH 4+ glutamatdehydrogenase NAPH + H+ NADP+

+

COOH3N C CH2 CH2 COOglutamate H

Thc cht y l qu trnh xy ra 2 giai on: lc u amoni kt hp vi ketoglutarate, sau l vic chuyn hydro t NADH hay NADPH n Glutamate.
O H C C H2 COO H N

H2 C
-OOC

H 2O + NH4
+
-

H2 C
OOC C H2

C COO-

alpha-ketoglutarate
NAPH + H+ NADP+

H N

NH 3+

H2 C
-

H2 C
-

C C H2 COOH

C C H2 COO-

OOC

OOC

glutamate

iu quan trng l, trong qu trnh chuyn hydro to ra cu trc lp th ti v tr carbon . Enzyme glutamatedehydrogenase lin kt vi -ketoglutarate, lm cho hydro ch chuyn vo v tr ring bit, do trong phn ng ny ch to nn Lglutamate. Mt ion amoni khc s kt hp vi glutamate di tc dng ca enzyme glutamine synthetase hnh thnh nn glutamine. Phn ng ny c thc hin bi nng lng ca ATP.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

NH3+ O
-OOC

ATP

ADP
-OOC

NH3+ O HP H O OH OH OH

2-

H O glutamate

acyl-phosphate intermediate

NH3+ NH2
-OOC

NH3

Pi

H O glutamine

Qa trnh sinh tng hp glutamine ng vai tr rat quan trng trong vic iu khin cc qu trnh trao i cht. Cc enzyme glutamate dehydrogenase v glutamine synthetase u c tm thy trong cc sinh vt sng. Glutamate cng c th c to thnh t glutamine
COO+

COOH + C CH2 CH2 O NAPH + H+ NADP+ 2

+

COOH 3N C CH2 CH2 COOglutamate H

H3N

C CH2 CH2 C

O glutamine

NH2

COOalpha-ketoglutarate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.1.1.2. Proline

Hnh 2.1 : Cu trc ca Proline Cng thc phn t Khi lng phn t Mu sc v tnh ht m im nng chy C5H9NO2 115.13 g mol1 Tinh th khng mu, ht m 205 C

Kh nng ha tan trong nc Ha tan

Proline khc vi tt c cc amino acid khc ch nhm amin bc 1 v tr kt hp vi mch bn to thnh vng pyrrolidine, do proline l mt imino acid cha nhm amin bc 2. C ch sinh tng hp proline

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Hnh 2.2. Con ng sinh tng hp Proline t Glutamate hoc Ornithine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Hnh2.3. Mi quan h gia vi khun v ng vt c v trong vic sinh tng hp Proline Hu ht trong cc loi vi khun (nh l Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium) proline c tng hp t 1 tin cht amino acid L glutamate, qu trnh tng hp ny gm 4 bc. u tin di tc dng ca enzym glutamate kinase, ly 1 nhm phosphoryl t ATP to ra glutamate 5-phosphate. T y thc hin phn ng kh vi s c mt ca enzym glutamate dehydrogenase, s dng 1 NAD(P)H+H+ , nhm pirophosphate c gii phng, glutamate semialdehyde hnh thnh, ngay lp

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

tc cht ny c chuyn thnh tin cht 1-Pyrroline-5-carboxylase (P5C) m khng cn 1 enzym no . Bc cui cng P5C vi xc tc ca enzym Pyrroline5-carboxylate reductase (i hi 1 NAD(P)H+H+) proline c to thnh. 2.1.1.3. Arginine Arginine c to thnh t glutamate. Phn ng din ra nh sau:
COOH H 2N C CH2 CH2 COOH glutamate H Acetyl-CoA CoA transacetylase O HN C COOH C CH2 H ATP ADP O HN C COOH C CH2 H

CH3 CH2 COOH N-acetyl glutamate

CH3 CH2 OC P

N-acetyl-gama-glutamine phosphate

COOH NADPH NADP+ reductase O HN C C CH2 H glutamate alphaketoglutarate O transaminenase HN C

COOH C CH2 H

CH3 CH2 CHO N-acetyl-glutamic-gamasemialdehyde

CH3 CH2 NH2 N-acetyl-ornithine

N-acetylornithinease

COOH H 2N C H

(CH2)3 NH2 orthinine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Orthinine tip tc tham gia chu trnh Ure to thnh Arginine

COOH H2N C H transcarbamylase carbamylphosphate H2N COOH C H

(CH2)3 NH2 orthinine

(CH2)3 NH C NH2 Citrulline ATP + Aspartate argininosuccinate synthetase O

AMP + PPi

COOH COOH H 2N C H argininosuccinase H2N C H

(CH2)3 NH COOH N C CH2 COOH argininosuccinate H

(CH2)3 HN C NH2 f umarate

C NH2

NH L-arginine

2.1.2. T oxaloacetate 2.1.2.1. Aspatate/ Asparagine a. Aspatate Aspartate c sinh ra bi s chuyn amin

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

COO+

COOH + O CH2 aspartate transaminase C COO+H 3N

COOCOOC C CH2 CH2 COOaspartate COOalphaketoglutarate H + CH2 O

H 3N

C CH2 CH2 COO-

glutamate

oxaloactate

b. Asparagine Asparagine c to thnh t aspatate di tc dng ca enzyme Asparagine synthetase. Gm 2 giai on Giai on 1: Hot ha nhm carboxylate ca aspatate bng ATP to nn aspartyl adenylate intermediate Giai on 2: Cp nucleophile ca ammoniac s tn cng vo nhm carbonyl c hot ha ca aspartyl-adenylate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

NH2 N O C O H C NH3+ aspartate O H2 C C O O N O O O P O P O O P O O O O H HO OH H H H N N

O O P O O

O P O N N O NH2

O O C H C NH3+ H H2 C

O C O

O O P O O H H H

N H

AMP O O C H C NH3+ H2 C O C NH2 Asparagine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Trong mt qu trnh khc, asparagine c th c to ra bi s chuyn mt nhm amin t glutamine n aspartate.

COO+H 3N

COOH +
+H 3N

COOH
ATP
+H 3N

COO H + O
+H 3N

C CH2 CH2 COO-

C CH2 CH2 C

C CH2 C NH 2

AMP + PPi

C CH2 CH2 COO -

aspartate

NH2 glutamine

asparagine

glutamate

2.1.2.2. Methionine
CH 2 OH CoA S C O H2 C O C CH 2 NH2 CH 2 COOH O

CH2
HC COOH NH2

CH2
CH 2 COOH

transaminase

CH2
HC COOH

homoserine

succinyl CoA

o-succinyl homoserine cistationine gama-synthetase

S CH 2

CH 3

SH

H 2C

CH2 HC NH2

+ CH2-THF

CH 2

cystationase

CH 2
HC NH2

CH2
HC COOH NH2

CH2
HC COOH NH2

COOH

COOH

cystationine methionine homocystein CH2-THF: N5-methyltetrahydrof olate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.1.2.3. Threonine
COOH COPO3H2 CHO
+ NADPH + H+ NADP

CH2
HC COOH NH 2

ATP

ADP

CH 2
HC COOH

CH2
HC COOH NH2

NH2

aspatate

beta-aspartylphosphate

aspartate-betasemialdehyde NADPH + H + NADP+

CH 3 H H2 N

H2 C

CH 2OH

C
C

OH H H2N

CH 2
CH COOH

CH 2
H2N CH COOH

Pi

ATP

ADP

COOH

L-threonine

homoserinephosphate

homoserine

2.1.2.4. Lysine a. Qu trnh sinh tng hp L-Lysine vi khun

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

OOC COO H3N H L-aspartate

-

ATP

ADP

OOC H 3N H

O P

aspartokinase

O O O L-aspartat-4-yl phosphate NADPH beta-aspartate semialdehyde dehydrogenase NADP+

-OOC

OOC H 3N HHO H

COO -

pyruvate H H

dihydropicolinate synthetase

H3N

L-aspartate semialdehyde

OH NADPH
-

OOC H N COO-

NADP+ OOC H N COO(S)-2,3,4,5tetrahydropyridine -2,6-dicarboxylate succinylCoA CoA

OOC H N COO(S)-2,3-dihydropyridine -2,6-dicarboxylate

OOC H H

COO NH 3

OOC H

COO -

HN

HN

COON-succinyl-L-2,6-diaminoheptanedionate

COON-succinyl-L-2amino6-oxoheptanedionate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

H 2O succinate
-OOC

COOH NH3

-OOC

COOH NH3

epimerase H NH3

NH3 LL-2,6-diamino heptanedionate

meso-2,6-diamino heptanedionate

CO2
-OOC

NH3

NH3 L-Lisine

b. Qu trnh sinh tng hp L-Lisine nm men v nm si

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

C CH3

CoA

COOH CH2

COOH CH

C CH2 CH2

COOH

-CoASH 2

HO

C CH2 CH2

COOH

-H2O

C CH2 CH2

COOH

COOH 1

COOH 3

COOH 4 5

COOH H C CH2 CH2 CH2 COOH 10 11 COOH H C CH2 CH2 CH2 O NH2 NH 2 9

COOH C CH2 CH2 CH2 COOH 8 O H

COOH C C CH2 CH2 COOH O COOH 7 HO H

COOH C C CH2 CH2 COOH 6 H COOH

COOH H C CH2 CH2 CH2 OH NH2 H

COOH C CH2 CH2 CH2 H2C H N 14 NH2 COOH CH2 CH2 CH2 H

COOH C CH2 CH2 CH2 H2C 15 NH2 NH2

C O AMP H2C 12 13 1. acetyl-CoA-alpha-ketoglutarate 2. homocittrate synthetase 3.homocitrate 4.homoacotinate 5.hidrase

6. homoizocitrate 7. dehidrogenase 8. alpha-ketoadipate 9. transaminase 10. alpha-aminoadipate

11. synthetase 12. adenilyl-alpha-aminoadipat 13. alpha-aminoadipic semialdehyde 14. saccaropin 15 Lisine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.1.3. T pyruvat 2.1.3.1. Alanine Alanine c to thnh t phn ng chuyn amin t pyruvate v glutamate.
COOH HC CH2 CH2 COOH glutamate pyruvate alanine NH2 + H3C C O COO
-OOC

H3C

H C NH2

CH2 COO
-

H2 C C
- OOC

alpha-ketoglutarate

2.1.3.2. Valine v Leusine Valine.

Hnh 2.4: Cu trc ca Valine Tn khc : 2-amino-3-methylbutanoic acid. Cng thc phn t : C5H11NO2 Khi lng phn t : 117.15 g/mol Khng phn cc, k nc.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Leucine.

Hnh 2.5. Cu trc ca Leucine Tn khc : 2-Amino-4-methylpentanoic acid Cng thc phn t : C6H13NO2 Khi lng phn t : 131.17 g mol1 Khng phn cc Valine v Leucine l 2 trong nhng amino acid thit yu mch nhnh m c th ng vt v con ngi khng th t tng hp c m phi c cung cp thng qua thc n hng ngy.

Hnh 2.6. Con ng sinh tng hp Valine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Valine v leucine u i t piruvat v cng chung sn phm trung gian l acid cetoisovalerianic. Ph thuc vo tiaminpirophosphat m piruvat s ngng t vi 2 mnh cacbon to nn acid acetolactic. Di tc dng ca enzim isomeroreductaza acetooxyacid (enzyme ny i hi phi c mt Mg2+ v piridinnucleotit dng kh) acid acetolactic s chuyn v pinacolic to nn acid , dihydroxyisovalerate. Acid dihydroxyvalerianic ny di s xc tc ca enzyme dehydratase ca dihydroxyacid s b mt nc v to thnh acid ketoisovalerianic, sau khi chuyn amin ha acid ny di s c mt ca enzym valine aminotransferase s trc tip bin thnh valine.

Hnh 2.7. Con ng sinh tng hp Leucine Acid cetoisovalerianic ngng t vi acety ScoA to nn acid isopropylmalate. Acid ny s bin thnh aicd isopropylmalate nh enzym isopropylmalate isomerase. Tip tc isopropylmalate s b kh cacboxyl ha bng cch oxy ha to thnh ketoisocaproate, v gii phng CO2. Bc cui

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

cng ketoisocaproate thc hin chuyn amin ha nh enzym leucine amonitransferase to thnh leucine. 2.1.3.4. Isoleusine Isoleucine l nhng acid amin c mch nhnh. Isoleucine th c to thnh t treonin. Di tc dng ca enzym reonindehydratase, treonin b chuyn ha thnh cetobutirat. - cetobutirat s ngng t vi acetaldehid hot ha (acetaldehid hot ha l nhm oxyetyl ca cht oxyetyl 2 tiaminpirophosphat) to thnh acid aceto oxybutyric mt sn phm trung gian then cht trong vic tng hp isoleucine. S tng hp isoleucine t sn phm ny tin hnh rt ging s tng hp valine t acetolactat. Hnh nh, giai on cui cng trong phn ng chuyn amin ha, c hai phn ng lin tip nhau u ch c cng mt loi enzyme xc tc. Tuy nhin tng hp valine c hai transaminase u c s dng, cn tng hp isoleucine th ch c mt enzyme c s dng. iu gii thch hin tng l trong thin nhin c tn ti nhng bin chng cn isoleucine v khng cn valine. Cn ch rng isoleucine c ch hon ton enzyme treonindehydratase (ng hn l enzyme treonindezaminase) l enzyme xc tc phn ng u tin ca chui phn ng sinh tng hp isoleucine. S km hm treonindehydratase y mang tnh cht canh tranh v i lc cc enzyme vi L isoleucine ln hn vi L treonine 100 ln.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

COOH -NH2 HOH2C C H threonine NH2 H3C H2 C

COOH C O H 3C H2 C

COOH C C OH O

alpha-ketobutirate

CH3 alpha-aceto-alphahydroxybutirate +2H+ COOH -H2O H3C H H2 C C C CH3 alpha,beta-dihydroxy-betametyl valerat OH OH

COOH H H2 C C C CH3 L-Isoleucine NH2 OH +NH2 H2 C

COOH C H3C C CH3 alpha-keto-beta-metyl valerate O OH

H 3C

2.2. T phosphoglycerate
2.2.1.Serine Serine c to thnh t 2 giai on t 3 phosphoglycerate Giai on 1: oxi ha 3 phosphoglycerate Giai on 2: amin ha.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

COOH H

COOOH O P

C C
H

phosphoglycerate dehydrogenase NAD+ NADH + H+

H

C C
H

O O P

3-phosphoglycerat 3-phosphohydroxypyruvate glutamate phosphoserine amino tranf erase alpha-ketoglutarate

COO +

COOH
+

H3 N H

C C
H

phosphoserine phosphatase
OH

H3 N H

C C
H

H O P

Serine

3-phosphoserine

2.2.2. Glycine Glycine c tng hp t serine di tc dng ca enzyme serine hydroxymethyltransferase

COO+

H4 folate C C
H H OH

N 5, N10-methylene H4 folate
+H 3N

H 3N H

COO-

C
H

Serine

H 2O

Glycine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

a. Tetrahydrofolate Tetrahydrofolate gm c 3 nhm: pteridine, amino benzoate v 1 nhnh gm 1 hay nhiu phn t glutamate.
H 2N

H N H

N H
OH

HN OH N p-aminobenzoate H COON n H COOOOC H

Pteridine

Glutamate

ng vt c v, chng c th tng hp c 3 thnh phn trn nhng li thiu enzyme lien kt chng li vi nhau. Ngc li, vi sinh vt li c nhng enzyme lien kt cn thit v c th sn sinh ra folate. Di tc dng ca enzyme dihydrofolate reductase. Folate s b bin i to thnh THF.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

H 2N

N H2 C N H

HN

f olate NADPH + H+

NADP+
H 2N

H N H2 C N H

HN

dihydrofolate NADPH + H+

NADP+
H 2N

H N H2 C N H

HN

N H

tetrahydrofolate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Tetrahydrofolate l cht mang cc nhm 1 carbon, hnh thnh lin kt gia N5 v N10. Carbon ny c th 3 trng thi oxy ha khc nhau: methyl (CH3); methylene (-CH2); formyl (-CHO); formimino (-CHNH) hay methenyl (-CH=). Nhm mt carbon c vn chuyn bng THF c th bin i qua li:

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

H N Ser H 5 N H HN10 tetrahydrof olate formate + ATP Gly

H N H N H2C N

H+ + NADP+ NADPH

H N H N H3C HN

N5, N10-methylenetetrahydrof olate NADP+

N5-methyltetrahydrof olate

ADP +Pi

NADPH

H N H N H N HC O N10-f ormyltetrahydrofolate H N H N CH HN O

H N NH3 H N HC N HN N5, N10-methenyltetrahydrof olate

H N H N CH HN

N5-formiminotetrahydrofolate

N5-formyltetrahydrofolate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

lys HN O C OH
2-

O3PO

H 3N

CH CH2

N H O O C H C HN CH2 O

OH

BH

OH
2-

O3PO

N H H2C O O C CH HN O

OH
2-

O3PO

N H

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

O O C CH HN H B O

O C CH2 HN

OH
2-

OH
2-

O3PO

O3PO

N H

lys O HN + C CH2 NH3 N H Glycine O

OH
2-

O3PO

b. SAM THF c th c cha nhm methyl, nhng nng lng ca chng li khng thc hin ton b qu trnh chuyn nhm metyl, do cht cha nhm methyl c s dng nhiu nht l S-Adenosylmethionine (SAM). SAM c hnh thnh bng s chuyn nhm adenosyl t ATP sang methionine. Nhm methyl ca methionine s c hot ha bi in tch dng trn nguyn t S, chnh in tch dng ny lm cho qu trnh chuyn nhm methyl din ra mnh m hn so vi N5methyl-THF.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

NH2
- OOC - OOC

NH 3 + H

NH3

N H

+
S

ATP
S H3 C H

N O H

Pi + PPi

H3 C OH OH

Methionine

S-Adenosylmethionine (SAM) RH RCH3 + H+

NH 2
-OOC -

OOC

NH3 + H

NH 3+ N

H N O N

Adenosine
SH

H 2O

Homocystein

OH

OH

Homocystein c chuyn thnh methionine bng cch chuyn nhm methyl t N5-methyltetrahydrofolate di tc dng ca enzyme homocystein methyltransferase.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

H H
-

NH 3 +

H N H

NH 3 +

OOC

H N H

OOC

+
N CH 3 SH HN H 3C S

+
N H HN

Homocystein

N5-methyl-tetrahydrof olate

methionine

tetrahydrof olate

2.2.3. Cystein Homocystein l sn phm trung gian trong qu trnh sinh tng hp cystein. Serine v homocystein kt hp vi nhau di tc dng ca enzyme cystathionine synthase to thnh cystathionine. Sau thc hin phn ng kh amin cystathionine di tc dng ca enzyme cystathionine -lyase. C hai enzyme ny u c cofactor l pyridoxal phosphate.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

H3N C H

CH 2 C H2 COOHomocystein

SH + H

COOH2 C C NH 3+ Serine

OH

Cystathionine beta-synthase H2O

COO+

H3N C H

H2 C
C H2

S C H2

C NH3 +

cystathionine

COO-

cystathionine gama-lyase H2O

H3N C H

H2 C
SH

OOC

C O

H2 C CH 3

COO cystein

alpha-ketobutyrate

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.3. Cc amin thm

Tryptophan

Hnh 2.8: Cu trc ca tryptophan Tn khc : 2-Amino-3-(1H-indol-3-yl)propanoic acid Cng thc phn t : C11H12N2O2 Khi lng phn t : 204.23 g mol1 Phenylalanine

Hnh 2.9: Cu trc ca tryptophan Tn khc : 2-Amino-3-phenylpropanoic acid Cng thc phn t : C9H11NO2 Khi lng phn t : 165.19 g mol1

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Tyrosine

Hnh 2.10: Cu trc ca tyrosine Tn khc : 2-Amino-3-(4-hydroxyphenyl)propanoic acid Cng thc phn t :C9H11NO3 Khi lng phn t : 181.19 g mol1 * C ch sinh tng hp Con ng sinh tng hp cc amino acid thm phenylalanine,

tyrosine v tryptophan cng c chung nhiu tin cht trung gian

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Hnh 2.11. Con ng chung tng hp cc amino acid thm (phenylalanine, tyrosine, tryptophan). (Theo: Prescott v cs, 2005)

Hnh 2.12 .Mi lin h ca qu trnh tng hp cc acid amin thm v vitamine E. coli

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.3.1. Tng hp ra tin cht trung gian Chosrimate.

Hnh 2.13. Con ng sinh tng hp Chorismate Sinh tng hp Chorismate ch c th xy ra thc vt v vi khun, khng xy ra ng vt Bc u tin l s ngng t ca phosphoenolpyruvate ( l mt glycolytic trung gian ) vi erythroe-4-phosphate ( mt pentose phosphate ). Kt qu l to ra mt hp cht 7 carbon m u cho chui oxy ha tip theo, hp cht ng ny sau b mt i nhm phosphoryl, thc hin vic ng vng to thnh 3-dehydroquynate.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Phn ng tip theo l s loi i 1 phn t nc to ra 3-dehydroshikimate. S kh NADPH +H+ thnh NADP+ t 3-dehydroshikimate thnh shikimate. Tip thep shikimate ly 1 nhm phosphoryl t ATP to thnh shikimate-3-phosphate v ADP. Mt phn t phosphoenolpyruvate th hai s c s dng ngng t vi shikimate-3-phosphate, ti y khi kt hp vi shikimate-3-phosphate n s b mt i nhm phosphoryl ca n, to thnh 3-enolpyruvyl-shikimate-3-phosphate ri to ra chorismate . y chnh l tin thn chung ca c 3 acid amin thm. Cc enzyme tham gia vo cc phn ng gm: 1) 3-deoxy-D-arabino-heptulosonate 7-phosphate synthetase (DAHP synthetase ) 2) 3-dehydroquinate synthetase. 3) dehydroshikimate reductase. 4) shikimate kinase. 5) 5-enolpyruvylshikimnate 3-phosphate synthetase. 6) chorismate synthetase .

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

2.3.2. Sinh tng hp Phenylalanine v Tyrosine

Hnh 2.14. Con ng sinh tng hp Tyrosine v Phenylalanine Chorismate t chuyn thnh prephenate, y l tin cht trc tip ca 2 acid amine thm phenylalanine v tyrosine. T cht ny n c th dehydro ha v decarboxyl ha to thnh phenylpyruvate hoc theo 1 cch khc, n c th kh decarboxyl to thnh -hydroxylphenylpyruvate. Ti y nhng -ketoacid c thc hin transaminate to thnh phenylalanine v tyrosine. Cc enzym tham gia phn ng bao gm : 1) choris-mate mutase. 2) prephenate dehydratase. 3) phenylalanine transaminase. 4) prephenate dehydrogenase. 5) tyrosine transaminase.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Ngoi ra Tyrosine cn c th c tng hp t phenylalanine nu nhu cu v tyrosine b thiu, lc ny mt na lng phenylalanine s b ly i thc hin vic sn xut tyrosine. Phn ng ny thc hin nh enzym phenylalanine hydroxylase, y l mt enzyem c chc nng hn hp, mt nguyn t oxy c a vo trong nc, cc nguyn t khc c a vo nhm hydroxyl ca tyrosnie. Cht kh l mt cofactor tetrahydrobiopterin, nh enzym dihydropteridine reductase , dihydrobiopterin to thnh li c tng hp nn tetradrobiopterin tip tc thc hin vic tng hp tyrosine

Hnh 2.15. Con ng tng hp Tyrosine t Phenylalanine 2.3.3. Sinh tng hp Tryptophan

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Hnh 2.16. Con ng sinh tng hp Tryptophan Chorismate nhn 1 nhm amin t glutamine thy phn mch bn, to thnh anthranilate v gii phng 1 pyruvate. Sau anthranilate ngng t vi 5phosphoribosyl-1-pyrophosphate (PRPD), mt dng hot ha ca ribose phosphate. PRPD cng l mt cht trung gian quan trng tng hp histidine, cc nucleotide purine, v cc nucleotide pyrimidine. Nguyn t C v tr C-1 ca ribose 5phosphate lin kt vi nguyn t nit ca anthranilate trong phn ng , v gii phng ra 1 pyrophosphate. Mt na ribose ca phn t phosphoribosylanthranilate chu s sp sp li to thnh 1-(o-carboxyphenylamino)-1-deoxyribulose 5phosphate. Sau ngay lp tc thc hin dehydrate v decarboxylate to thnh

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

indole-3-glycerol phosphate, ri thc hin ct 3-glycerol phosphate cho ra indole. Tip theo Indole phn ng vi serine to thnh tryptophan. Trong bc cui cng phn ng c xc tc bi enzym synthetase tryptophan. Mch bn ca indole-3-glycerol phosphate c ly ra to thnh glyceraldehyde 3-phosphate v thay th vo l b khung carbon ca serine. Cc enzym tham gia phn ng gm : 1) anrailaie synthetase 2)anthranilate-5'-phosphoribosyl-1-pyrophosphatephosphoribosyl transferase (PR transferase) 3) N-(5'-phosphoribosyl)-anthranilate isomerase 4) indoleglycerol phosphate synthetase 5) tryptophan synthetase

2.4. Histidine
The synthesis of histidine is long and complex and its pathway is intertwined with nucleic acid biosynthesis (specifically purine). y l con ng tng hp chung cho tt c cc vi sinh vt c kh nng tng hp histidine. u tin l phn ng tch nhm Ribose t phosphoribosyl pyrophosphate (PRPP) v chuyn sang Imadiazoleglycerol phosphate. Khi lin kt imadiazole c hnh thnh, glutamate s cho nhm v amin mi c to thnh s tip tc b oxi ha thnh histidine giai on cui ca qu trnh. Ngun nng lng s dng trong trng hp ny l ATP (trong trng hp ny, cc nguyn t ca phn t ATP s tr thnh thnh phn ca acid amin v pyrophosphate c hnh thnh t phosphoribosyl pyrophosphate.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

Hnh 2.16. Sinh tng hp Histidine

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

CHNG 3: KT LUN
Ngy nay cc acid amin c s dng rng ri trong y hc, trong chn nui v trong cng ngh thc phm. Trong s 20 acid amin thng gp trong phn t acid amin m c th ngi v ng vt khng th t tng hp c m phi a t ngoi vo qua thc n . Trong khi , vi sinh vt li c kh nng tng hp ht 20 acid amin . Mc d c th sn xut bng con ng tng hp ha hc nhng nhc im ca phng php ny l sn sinh ra c hai dng ng phn quang hc. Tri li con ng tng hp sinh hc ch sn xut ra cc L-acid amin l dng hay gp trong cu trc ca protein trong c th sng.

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Cc con ng sinh tng hp acid amin vi sinh vt

Nhm 1

TI LIU THAM KHO

1. Nguyn Vn Kim (ch bin), Nguyn Vn Knh, Nguyn Vn Mi, Ha sinh ng vt, BGD-T i Hc Nng Nghip H Ni, 2005. 2. Nguyn nh Quyn, Nguyn Ln Dng, S dng nng lng trong sinh tng hp vi sinh vt. 3. Frank Gibson and James Pittard, Pathways of Biosynthesis Aromatic Amino Acids and Vitamins and Their Control in Microorganisms, Bacteriological Reviews, Dec.1968, p 465 492 4. http://www.blc.arizona.edu/courses/bioc462b/grimes/nitrogen06/aa.cfm 5. http://www.biocarta.com/genes/catMetabolism.asp 6. http://lecturer.ukdw.ac.id/dhira/Metabolism/aminoacids.html 7. http://www.themedicalbiochemistrypage.org

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