Bai Giang Cong Nghe Hoa Sinh 2010

8/6/2019 Bai Giang Cong Nghe Hoa Sinh 2010

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I HC THI NGUYNTRNG I HC KHOA HC

KHOA KHOA HC S SNG--------------------------

BI GING:

CNG NGH HO SINH

Ngi son: ThS. Trnh nh Kh

THI NGUYN - 2010


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CHNG 1

CNG NGH SN XUT KHNG SINH V KHNG SINH BN

TNG HP

1. I CNG V KHNG SINH

2. CNG NGH SN XUT KHN SINH PENICILLIN

3. CNG NGH SN XUT -LACTAM BN TNG HP


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1. I CNG V CHT KHNG SINH

1.1. LC S NGHIN CU

Thut ng cht khng sinh ln u tin c Pasteur v Joubert (1877) s dng

m t hin tng km hm kh nng gy bnh ca vi khun Bacillus anthracis trn

ng vt nhim bnh.

Babes (1885) nu ra nh ngha hot tnh khng khun ca mt chng l c tnh

tng hp c cc hp cht ho hc c hot tnh km hm cc chng i khng.Nicolle (1907) l ngi u tin pht hin ra hot tnh khng khun ca Bacillus

subtilis c lin quan n qu trnh hnh thnh loi bo t.

Gratia v ng nghip (1925) tch t nm mc mt ch phm c th s dng iu

tr hiu qu cc bnh truyn nhim trn da do cu khun v ng l ngi u tin

xy dng hon thin phng php tm kim v pht hin vi sinh vt sinh tng hpcht khng sinh trong t nhin.

Nm 1929 thut ng cht khng sinh mi c Alexander Fleming m t mt cch

y v chnh thc trong bo co chi tit v penicillin


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1.2. KHI NIM CHT KHNG SINH

Theo cc nh sinh hc, khng sinh l nhng hp cht ho hc do vi sinh vttit ra c tc dng c ch s pht trin hay tiu dit mt cch chn lc mt

nhm vi sinh vt xc nh hay c tbo ung th nng thp.

Cn cc nh ho hc th mun nh ngha khng sinh sinh phi bao hm c

cc cht tng hp bng ho hc c tc dng dit khun nh cc cht thuc

quinolon.

Cht khng sinh (antibiotic) l cc cht ho hc xc nh, khng

c bn cht enzyme, c ngun gc sinh hc vi c tnh ngay

nng thp c kh nng c ch mnh m hoc tiu dit c

cc vi sinh vt gy bnh m vn m bo c an ton cho ngi

hay ng vt c iu tr.


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Tn khng sinh Vi sinh vt sn xut khng sinh Hot ph

Penicillin Penicillium chrysogenum Khng khun

Steptomycin Str. griseus Khng khun

Tetracyclin Str. aureofaciens Khng khunChloramphenicol Str. venezuela Khng khun

Oxytetracyclin Str. rimosus Khng khun

Cephalosporin Cephalosporium acremonium Khng khun

Actinomycin D Str. antibioticus Khng ung th

Bleomycin Str. verticillum Khng ung th

Daunorubicin Str. peuceticus Khng ung th

Mitomycin C Streptomyces sp. Khng ung th

Kanamycin Str. kanamyceticus Khng khun

Nystatin Str. noursei Khng nm

Fumagillin A. fumigatus Khng protozoa

Griseofulvin P. griseofulvum Khng nm

Nistin Streptococcus sp. Bo qun thc phm

Natamycin Streptococcus sp. Bo qun thc phm

Rifamycin Nocardia sp. Khng lao

Polymycin B Bacillus polymyxa Khng khun

Gentamycin Micromonospora sp. Khng khun


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1.3. C CH TC DNG CA CHT KHNG SINH

Lm ri lon cu trc thnh tbo

Ri lon chc nng iu tit qu trnh vn chuyn vt cht qua mng tbocht

Lm ri lon hay kim to qu trnh sinh tng hp protein

Ri lon qu trnh tng hp DNA

Hoc tng tc c hiu vi nhng giai on nht nh trong cc chuyn

ho trao i cht.

nh gi hiu qu ng dng ca cht khng sinh:

hot tnh khng sinh c hiu, ph khng sinh

p ng phn hi ca mm bnh i vi khng sinh

phn ng ph ca thuc, ng cp thuc

s chuyn ho thuc trong c th bnh nhn, ng hng o thi thuc

tng tc qua li gia cc loi thuc khi s dng phi hp.


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1.4. CHC NNG SINH HC CA CHT KHNG SINH

Vic tng hp cc cht khng sinh nhm to ra u th pht trin cnh

tranh c li cho chng sinh khng sinh, nh chng c th tiu dit hay

km hm c s pht trin ca cc loi khc cng tn ti v pht trin

trong h sinh thi cc b .

Vic tng hp cht khng sinh l mt c tnh cn thit v m bo cho

kh nng sng st cao cho chng vi sinh ra cht khng sinh trong t nhin,

nht l i vi cc loi c bo t.


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1.5. IU CHNH SINH TNG HP CHT KHNG SINH

Tuyn chn v to ra cc chng cng nghip siu tng hp cht khng sinh

Ti u ho thnh phn mi trng, thit b ln men v iu kin vn hnh

qu trnh ln men.

1.5.1. Tuyn chn v to ra cc chng cng nghip siu tng

hp cht khng sinhTri qua 6 giai on c bn:

Phn lp t t nhin

Nghin cu x l to cc bin chng siu tng hp c hot lc cao

Tuyn chn s b Tuyn chn li thu cc chng c hot tnh cao quy m phng th nghim

Th nghim v tuyn chn li trn quy m sn xut th nghim pilot

Th nghim v chn lc li cc chng ph hp vi iu kin ln men

sn xut ln cng nghip.


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1.5.2. Ti u ho thnh phn mi trng, thit b ln men v iu kin vn hnh qutrnh ln men

ngun cacbon thng c la chn l: cc loi bt v ht ng cc, cm

m, cm go, v khoai ty, r ng, cc loi ng, dextrin, glyxerin, axit

axetic, dch thu phn g,...

Ngun nit c th l: bt u tng, nc chit ng, cao nm men, nc

chit nm men, pepton, cc loi mui v c cha nit,...

Cc nguyn t khong a lng nh: P, S, Mg, Fe, Ca, K, Na. Cc nguyn t

vi lng nh: Cu, Zn, Co, Mo,... v cc cht kch thch sinh trng.

Khai thc hiu qu tc ng ca cc yu t khc trong mi trng

nh: nhit ln men ti u, pH, nng oxy, th oxy ho - kh,

cng sc kh, cng khuy trn dch ln men,...


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1.6. CC BC C BN TO CHT KHNG SINH DNG TRONG Y HC

Bc 1: Phn lp vi sinh vt sinh khng sinh di dng thun khit. Chn lc chng chot ph khng sinh mong mun. nh tn chng vi sinh vt tuyn chn c.

Bc 2: Nui cy vi sinh vt sinh khng sinh trong thit b ln men dung tch t 5 100 lt.

Chit xut, tinh ch thu khng sinh tinh khit, xc nh cu trc ho hc, th s b v

c tnh v mt vi hng s l ho ca khng sinh tm c.

Bc 3: Nghin cu c im sinh l, sinh ho ca vi sinh vt sinh khng sinh, tm mitrng ti u nui cy. t bin ci to ging bng k thut di truyn c in v k

thut gen thu c chng ging c nng sut cao, n nh.

Bc 4: Nghin cu dc l ca khng sinh tm c, tc dng iu tr ca khng sinh

bng cch gy bnh thc nghim trn ng vt thnghim.

Bc 5: Nghin cu sn xut khng sinh trn qui m cng nghip bao gm: mi trng ln

men, cc thng s k thut ca qu trnh ln men, phng php chit xut v tinh ch

t tiu chun dng lm thuc,...

Bc 6: Nghin cu kinh tv th trng


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1.7. PHNG HNG PHT TRIN CNG NGH SN XUT KHNGSINH

Nghin cu sn xut thm cc ch phm bn tng hp, m rng ph iu tr v mang li ccc tnh u vit hn. Nghin cu cc hot tnh mi ca khng sinh nh: khng nm, khng

virus, khng sinh c kh nng km hm s pht trin ca cc khi u,...

Hon thin cc phng php phn lp, tuyn chn, to cc bin chng siu tng hp, iu

chnh ng hng cc qu trnh trao i cht v ng hng qu trnh ln men, p dng

cc cng ngh tch chit v tinh ch mi, hon thin v p dng cc k thut phn tch hin

i (c bit l k thut in vivo assay systems),... trin khai sn xut cng nghip cccht khng sinh.

Tip tc tm kim pht hin cc cht khng sinh mi, bao gm c m rng tm kim pht hin

kh nng sinh tng hp cc cht khng sinh sang cc nhm sinh vt khc

Sn xut cc ch phm iu tr vi s phi hp nhiu loi thuc, m rng kh nng ng dng

v iu tr ca cc cht khng sinh bit; ng thi gp phn lm gim nguy c khng

thuc v hn ch bt cc hiu ng khng mong mun ca mi cht khng sinh.

M rng thm kh nng ng dng cc cht khng sinh vo cc lnh vc kinh tv khoa hc

khc nh: ng dng trong chn nui, bo qun thc phm, nng nghip, bo v mi trng,

nghin cu khoa hc,...


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2. CNG NGH SN XUT KHNG SINH PENICILLIN

2.1. GII THIU KHNG SINH PENICILLIN

Hnh 1.1. Cu trc vng -lactam v khung penam

Hnh 1.2. Cu trc ca penicillin

-lactam Penam


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Bng 1.3. Cu trc phn t ca mt s penicillin t nhin

Gc R Tn gi gc Tn penicillin

C6H5-CH2- Benzylpenicillin Penicillin G

C6H5-O-CH2- Phenooxymethylpenicillin Penicillin V

CH 3-CH2-CH=CH-CH2- 2-pentenylpenicillin Penicillin F

CH3-(CH2)4- n-amylpenicillin Dihydropenicillin F

CH3-(CH2)6- Hepthylpenicillin Penicillin K

HOOC-(NH2)CH(CH2)3- 1-amino-1-carboxyl-buthylpenicillin

Penicillin N

HO-C6H5-CH2- p-hydroxybenzylpenicillin Penicillin X


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C chtc dng ca h khng sinh -

lactam


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Cu to thnh t bo vi khun?

Gram dng Gram m


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C ch tc dng


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2.2. C CH SINH TNG HP PENICILLIN NM MC P.CHRYSOGENUM

Hnh 1.3. S c ch tng hp penicilin nm mc P. chrysogenum

H2N CH C

(CH2)3

OH

O

C

OH

O

H2N CH C

CH2

OH

O

SH NH2

CH

C

CH

OH

O CH3

CH3

CH

H2N

HOOC

(CH2)3 CONH

N

SH

O

CH

H2N

HOOC

(CH2)3 CONH

N

S

COOHO

RCONH

N

S

COOH

O

H2N

N

S

COOH

O

Isopenicillin N

Penicillin N

Phenylacetic acid

Benzylpenicillin(Penicillin G)

axit aminoadipic Cystein Valin

aminoadipyl- cystein - valin

6-APA(6 amino penicillinic acid)

axit aminoadipic (H

ng1)

(H ng 2)

H n

g3


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2.3. QUY TRNH LN MEN SN XUT PENICILLIN TRONG CNGNGHIP

2.3.1. c im chung

Theo cng ngh ln men ca hng Gist-Brocades (H Lan) th ton b

dy chuyn sn xut penicillin c th phn chia lm 4 cng on chnh

nh sau:

Ln men sn xut penicillin t nhin (thng thu penicillin V hoc G).

X l dch ln men tinh ch thu bn thnh phm penicillin t nhin.

Sn xut cc penicillin bn tng hp

Pha ch cc loi thuc khng sinh penicillin thng mi


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2.3.2. Chun b ln men

a. Ging, bo qun v nhn ging cho sn xut

Ging cng nghip P. chrysogenum c bo qun lu di dng ng

kh, bo qun siu lnh -70oC hoc bo qun trong nit lng.

Ging t mi trng bo qun c cy chuyn ra mi trng thch hp

hot ho v thu bo t. Dch huyn ph bo t c cy chuyn tip

sang mi trng bnh tam gic, ri thit b nhn ging nh, thit b nhn

ging trung gian,... v cui cng trn thit b nhn ging sn xut.


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b. Chun b mi trng v thit b ln men

Cc thnh phn mi trng thng c cn ong ri pha ch ring rtrong cc thng cha ph hp, sau thanh trng gin on 121oC (hay

thanh trng lin tc khong 140-146oC) hoc bng phng php siu

lc ri mi bm vo trong thng ln men.

Thit b ln men trc khi a vo s dng phi c v khun, thng

c thanh trng bng hi nc p sut 2,5-3 at trong thi gian 1-3h.

Sau khi kh trng xong thit b c a vo s dng ngay. Khng kh

thng c kh khun s b bng nn on nhit, sau c lc kh

khun bng cch lc qua mng v khun hoc siu lc.


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Bng 1.4. Thnh phn mi trng ln men c bn ln men sn xutpenicilin

Lactose 20-50 kg/m3

Glucose 0-10 kg/m3, b sung gin on hoc lin tc trong qu trnh ln

men

Dch chit ng c c 15-50 kg/m3

Cc khong cht

NaNO3 0-5 kg/m3; Na2SO4 0-1 kg/m

3; CaCO3 0-10 kg/m3; KH2PO4 0-4 kg/m

3;

MgSO4.7H2O 0-0,25 kg/m3; MgSO4 0-0,02 kg/m

3; ZnSO4 0-0,04 kg/m3

Tin cht to nhnh b sung lin tc theo nhu cu. Theo l thuyt nhu cu

v phenylacetat l 0,47g/g penicilin G hoc 0,5g phenooxyacetat/gpenicillin V

Cht chng to bt b sung lin tc theo nhu cu


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2.3.3. K thut tin hnh ln men

k thut ln men chm c p dng trong hu ht cc c s sn xut penicillin v

thng c vn hnh theo phng php ln men bn lin tc gm phng nln men gin on theo m c b sung lin tc (hoc bn lin tc) mt hay mt vicu t kt hp vi phng n tun hon li mt phn h si ca m ln men

trc.

Qu trnh ln men c vn hnh theo hai pha. Pha u nui thu sinh khi

trong khong 2-3 ngy, pha sau ln men thu sn phm. Nhit ln men

pha u thng khng ch 30oC, pha sau t 22-25oC. Tc sc kh v

khuy trn c iu chnh duy tr nng oxy ho tan khong 30%.

Dch ln men ban u thng c pH khong 6,5-6,8; trong qu trnh lnmen thng khng ch pH n nh trong khong 6,2-6,8. Nng NH4+

thng khng ch trong khong 0,3-0,4 kg/m3. Cht ph bt thng s

dng l cc loi du bo nh: m ln, du u tng, du vng,... Thi

gian ln men mi m thng ko di trong khong 144-180h.


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2.4. X L DCH LN MEN, TINH CH THU PENICILLIN 2.4.1. Lc

dch ln men Thng thng, ch cn tin hnh lc mt ln trn thit b lc ht kiu bng ti

hoc kiu thng quay ri lm lnh dch ngay chuyn sang cng on tip theo.

Phn sinh khi nm c ra sch, sy kh v s dng ch bin thc n gia

sc.

2.4.2. Trch ly

Penicillin thng c trch ly dng acid ra khi dch lc bng dung mi

amylacetat hoc butylacetat pH=2-2,5; nhit t 0-3oC. Qu trnh trch

ly c thc hin trong thi gian rt ngn. Dch lc c bm trn ngthi vi dung dch H2SO4 hoc H3PO4 long c b sung thm cht chng

to nh v bm song song cng vi dung mi vo thit b trch ly.

T l dch lc: dung mi thng chn trong khong 4-10V dch lc/1V dung

mi.


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2.4.3. Ty mu ty mu ngi ta thng b sung trc tip cht hp ph vo dung mi cha

penicillin sau trch ly. Cht hp ph ph bin thng l than hot tnh. Sau than

hot tnh c tch v ra li bng s dng thit b lc kiu bng ti hoc thng

quay. Phn than sau lc tin hnh chng thu hi dung mi v x l hon nguyn

phc v cho m ln men sau

2.4.4. Kt tinh, lc, ra v sy thu penicillin t nhin

Penicillin G hoc V thng c kt tinh di dng mui bng cch bsung trc tip vo dung mi sau khi ty mu mt lng nh Kali acetathoc Natri acetat hoc tin hnh trch ly li sang dung dch KOH long ritin hnh c trong chn khng nhit thp. Sau b sung BuOH penicillin t kt tinh.

Sau khi kt tinh, tinh th penicillin c lc tch bng my lc ht thngquay. Khi sn phm t tinh sch theo yu cu (thng tinh khitkhng di 99,5%) tin hnh lc tch tinh th. Sau , ra v lm kh sb bng dung mi k nc nh isopropanol hay butylalcohl, ht chn khng tch dung mi v sy bng khng kh nng n dng sn phm bt muipenicillin.


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3. SN XUT CC -LACTAM BN TNG HP T PENICILLIN G

3.1. NHU CU SN XUT CC PENICILLIN BN TNG HP3.1.1. Tc dng iu tr ca khng sinh thuc nhm -lactam

Tc dng iu tr chnh ca penicillin v cc khng sinh thuc nhm -lactam l do

cu trc ca penicillin c nhiu im gn gi vi dipeptid D-analin-D-analin, mt hp

phn cu trc lp peptidoglucan ca thnh t bo. Do , hot tnh cc enzyme tham

gia tng hp thnh t bo b bin i, cc enzyme nhn nhm c cht. Kt qu l

phn thnh t bo mi tng hp vi s nhm ln s khng c hnh thnh, lm

cho thnh t bo ca mm bnh chc tng phn hoc khng c thnh t bo, dn

n chng d dng b t phn trong mi trng v b h thng min dch ca c th

tiu dit.ng thi, cc khng sinh -lactam cn lin kt c hiu vi mt s protein trn

mng t bo cht. S lin kt ny lm bt hot v ho tan cc protein c hiu

trn mng, dn n lm mt hot tnh polymerase v ATPase. Ngoi ra, cc nh

khoa hc cn pht hin thy nh hng ca penicillin n cc chuyn ho

phospholipid trong t bo.


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3.1.2. Ti sao phi sn xut khng sinh bn tng hp

L do 1: Nhiu trng hp iu tr vi penicillin xut hin hin tng khng thuc.

Bin chng E. coli gram m v mt s bin chng B. subtilis khng thuc pht trin bnh

thng trong mi trng cha penicillin.

Nguyn nhn ca hin tng ny?

v hiu kh nng khng thuc, v nguyn l c th tin hnh nh thno?

L do 2: Trn phng din iu tr. Trong tng trng hp c th ngi thy thuc mong mun c nhng ch phm khng

sinh tng ng vi nhng c tnh khc nhau nh: thuc c ph khng khun rng hoc

ph khng khun hp hn, thuc bn vi enzyme -lactamase, thuc d hp thu, thuc

t phn ng ph, thuc o thi nhanh,... hay i vi bnh nhn cc loi thuc c th

ung thng an ton v d c chp nhn hn. Do , lun mong mun tm ra cc chphm penicillin mi vi c tnh a dng v c kh nng ch nh iu tr phong ph

hn.


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3.1.3. Nguyn tc to khng sinh mi

nguyn tc c th hon thin theo hng ln men trc tip vi cc tin cht

to nhnh khc nhau thu khng sinh mong mun hoc khng s dng

tin cht thu 6-APA lm nguyn liu tng hp cc penicillin khc nhau.

Con ng kinh t hn c v c trin khai trong sn xut ln l ch ln

men trc tip thu penicillin G hoc V lm nguyn liu t tng hp cc

khng sinh khc nhau bng con ng bn tng hp.


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3.2. SN XUT 6-APA V SN XUT PENICILLIN BN TNG HP

3.2.1. Sn xut 6-APA

Hnh 1.7. Cng thc cu to ca 6-APA

C hai con ng khc nhau tng hp 6-APA

Con ng th nht tin hnh ln men nhng khng b sung tin cht to

khng sinh nh s hnh 1.3.

Con ng hiu qu hn c hin nay l ln men sn xut penicillin G hoc V;

sau p dng phng php ho hc hay s dng enzyme penicillin-acylase

phn ct mch nhnh bn R.


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Con ng 1

H2N CH C

(CH2)3

OH

O

C

OH

O

H2N CH C

CH2

OH

O

SH NH2

CH

C

CH

OH

O CH3

CH3

CH

H2N

HOOC

(CH2)3 CONH

N

SH

O

CH

H2N

HOOC

(CH2)3 CONH

N

S

COOHO

RCONH

N

S

COOH

O

H2N

N

S

COOH

O

Isopenicillin N

Penicillin N

Phenylacetic acid

Benzylpenicillin(Penicillin G)

axit aminoadipic Cystein Valin

aminoadipyl- cystein - valin


axit aminoadipic (H

ng1)

(H ng 2)

H ng

3


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Hnh 1.8. S tng hp 6-APA bng con ng ho hc

Penicillin G

C6H5CONH

N

S

COOSi(CH3)3

O

C6H5CONH

N

S

COOSi(CH3)3

O

C6H5CONH

N

S

COOSi(CH3)3

O

H2N

N

S

COOHO


C6H5-CH2-COR +

(1)(2)

(2) (3) (4)

(4)

(CH3)3SiClPCl5

-400C

ROH

-400C

H2O

Con ng 2.1


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Hnh 1.9. S tng hp 6-APA bng enzyme penicillin-acylase

Con ng 2.2


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3.2.2. Sn xut penicillin bn tng hp

Bng 1.5. Mt s khng sinh

penicillin bn tng hp


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T 6-APA c th d dng to ra c nhiu ch phm penicillin bn tng hp khc

nhau bng c hai con ng: acyl ho trc tip t 6-APA theo phng php ho

hc hay s dng enzyme penicillin-acylase. Tuy nhin trong thc t thng p

dng ph bin con ng acyl ho trc tip theo phng php tng hp ho hc.

V d: Sn xut Ampicillin bn tng hp t 6-APA

Phng php A:


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Phng php B:


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3.3. SN XUT CEPHALOSPORIN BN TNG HP

3.3.1. Gii thiu cephalosporin

Hnh 1.10. Cng thc cu to vng -lactam v khung cephem

Hnh 1.11. Cng thc cu to chung ca h khng sinhcephalosporin v cephalosporin C

-lactam Cephem


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Cng thc cu to chung ca h khng sinhcephalosporin


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Khng tt Gram (+), rt km vi Gram (-)

Tng hot tnh khng Gram (-) nhngkm hn th h th 3

hot tnh khng Gram (+) km nhngkhng c Enterobactericeaebao gmc vi khun sinh -lactamase

M rng hot tnh khng hn th h 3 vbn vi -lactamase


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3.3.2. Sn xut cephalosporin


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Tng hp 7-ACA?


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Con ngha hc


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Con ng

enzyme


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Con ngha hc

Con ngenzyme


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Hnh 1.12. S tng hp 7-ACA t cephalosporin C


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T 7-ACA nu loi nhm acetoxy nhm thC3 th s nhn c 7-amino deacetyl

cephalosporinic acid (7-ADCA) bng phn ng hydrogen ho. Mt khc t 7-ADCA

bng phn ng brom ho v acetyl ho s nhn c 7-ACA.


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T 7-ACA v 7-ADCA bng con ng ho hc tng hp c cc khng sinh

cephalosporin bn tng hp th h 1, th h 2 v th h 3. Mt s quy trnh bn tng

hp cc cht khng sinh nh sau:


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CHNG 2

CNG NGH SN XUT VACCINE

1. KHI NIM CHUNG V VACCINE V MIN DCH

2. HNG CHIN LC TO VACCINE V PHN LOIVACCINE

3. C IM CA MT S LOI VACCINE

4. C SCNG NGH SN XUT VACCINE TH H MI


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1. KHI NIM CHUNG V VACCINE V MIN DCH

1.1. KHI NIM CHUNG

Yu t quyt nh min dch l g?

L thnh phn protein c bit c trn b mt ca tc nhn gy bnh hay

trn b mt ca ch phm vaccine ca chnh tc nhn gy bnh .

Thnh phn ny c gi l khng nguyn (antigen) do mt gen hay mts gen ca vi sinh vt m ho quyt nh tng hp ln. Nhng gen chu

trch nhim tng hp thnh phn protein c tnh kch thch min dch c

gi l gen khng nguyn.

Cc loi hnh min dch? Min dch dch th (humoral immunity)

Min dch qua trung gian t bo (cellular mediated immunity).


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Khng th l g?

khng th (antibody) l mt loi protein mi c chc nng bo vto nn min dch cho c th.

Bn cht khng th trong hai loi hnh min dch?

Trong min dch dch th, khng th ho tan vo h tun hon

hay trong dch c th nn gi l khng th th dch.

Trong min dch qua trung gian t bo th khng th khng phi

cht ho tan m chnh l cc t bo ca h min dch cm ngkhng nguyn m thnh khng th t bo. Chng khng phi

cht ho tan m l thnh phn hu hnh.


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Th no l vaccine?

Vaccine l ch phm sinh hc hoc bn sinh hc c con ngito ra v a vo c th gy min dch, tp dt cho c th thc

hin qu trnh p ng min dch chng li tc nhn gy bnh khi

trung xm nhp vo nhng ln sau .


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1.2. TIU CHUN CA VACCINE V QU TRNH P NG MINDCH

Tiu chun vaccine?

An ton l tiu chun nh gi khi s dng vaccine trn i tng c

hng phi khng c gy bnh hay gy phn ng c hi.

V trng l vaccine ch cha duy nht mt hay mt vi loi c chn lm

vaccine m khng b nhim tp cc loi khc.

Hiu lc l vaccine phi kch thch sinh min dch cho c th. Tnh hiu lcthc cht l mc biu hin gy min dch ca khng nguyn.


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c im qu trnh p ng mim dch ca c th?

Min dch c to ra nh s tham gia ca h thng min dch trung ng

v h thng min dch ngoi bin cng nh nhiu loi t bo c thm quyn

min dch khc.

H thng tu xng to ra cc loi t bo ngun, t sinh ra cc t bo c

thm quyn min dch: lympho B, lympho T, i thc bo v mt s t bo

chuyn bit khc.

c h tr v tham gia ca hch, lch, cc m lympho ng rut, ngh hp cng nh t bo tua v mt s thnh phn khc.


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2. HNG CHIN LC TO VACCINE V PHN LOI VACCINE

2.1. HNG CHIN LC TO VACCINE

Vaccine to nn theo cng ngh c truyn vaccine c in (classicalvaccine)

Vaccine to nn bng cng ngh sinh hc phn t hay vaccine th h

mi vaccine cng ngh gen (genetically engineered vaccine).

2.2. PHN LOI

* Cn c vo tnh cht sinh hc ca vaccine chia lm 3 loi:

- Vaccine v hot bng cc yu t l, ho hc.

- Vaccine sng: gm vaccine v c v vacine nhc c

- Vaccine phn t: c to ra bng k thut gen


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* Cn c vo ngun cung cp khng nguyn, xt di gc to

min dch cho c th chia lm 2 nhm:

-Vaccine sng: bao gm vaccine nhc c, vaccine v c va vaccine

nguyn c. Trong , vaccine nhc c c th thuc loi: qua vt ch

khng cm th, nhy cm nhit , thch ng nhit thp, xo gen c.

- Vaccine v hot: bao gm vaccine nguyn th v vaccine phn t.

Trong vaccine phn t c th c tinh ch t ngun vi sinh vt ch

trong thin nhin; sn xut, tinh ch bng k thut gen; cha protein tnghp bng k thut gen.


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3. C IM CA MT S LOI VACCINE

3.1. NHM VACCINE CUNG CP KHNG NGUYN SNG C

NHN LN3.1.1. Vaccine sng v c

L cc chng vi sinh vt v c c chn lc t trong qun th vi sinh

vt c trong t nhin. Chng hon ton khng cn hay chcn tnh gy

bnh rt yu i vi i tng c hng vaccine nhng vn c khnng to min dch rt tt cho c th.

3.1.2. Vaccine sng nguyn c

L loi vaccine cha cc chng vi sinh vt c c lc vi loi ny nhng

an ton hn i vi loi khc. Thc cht l loi vaccine v c i vi

loi l i tng c hng vaccine. Tuy nhin, chng vn gi nguyn

tnh cng c i vi loi thch ng m chng vi sinh vt c phn lp

lm vaccine.


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3.1.3. Vaccine sng nhc c

Hng th nht: Vaccine c th c to ra bng cch dng ho cht x l cc

chng vi sinh vt gy bnh lm vaccine. Tuy nhin ngy nay loi vaccine ny

khng c s dng v khng m bo tnh an ton.

Hng th hai: chn lc v x l nhng chng nhy cm vi nhit . Cc chngny khng hoc kh tm thy iu kin thch hp cc iu kin nhit bt

thng, do dn n s thch ng mt kh nng gy bnh nhng vn cn tnh

kch thch min dch.

Hng th 3: Lm thch ng mt s chng gy bnh iu kin nui cy lin tc

vi nhit thp (25oC). Loi ny c gi l vaccine thch ng iu kin lnh.

* Vaccine nhc c to ra qua tc dng l ho:


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* Vaccine nhc c to ra bng phng php tip truyn cin (Vaccine sng nhc c c in):

Phng php tip truyn c in l phng php tip truyn chng vi sinh vt

chn lm vaccine qua mi trng khng hoc t cm th nh tip truyn qua mi

trng tbo, qua trng c phi hay ngay trn ng vt khng hoc t cm th.

Qu trnh tip truyn thng c thc hin t hng chc n hng trm i lin

tc trong mi trng khng hoc t cm th (mi trng khng cung cp iu kin

ti u thc hin y chu k sng).

cc chng vi sinh vt phi c s bin i thch ng dn n s thay i nht nh

trong h gen biu hin tnh gy bnh.


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* Vaccine sng, ti t hp c vect dn truyn:

L loi vaccine c 2 thnh phn chnh: mt thnh phn l on DNA cha

gen khng nguyn a t bn ngoi v mt thnh phn l bn thn h

gen ca vect. H gen ca vect c nhim v tip nhn v biu th gen

khng nguyn trong nhng iu kin thun li.

* Vaccine Nucleic acid

Thc cht vaccine DNA chnh l s s dng DNA ca plasmid ti t

hp cha gen khng nguyn c ly t mt vi sinh vt no v c

ghp vo vect biu hin c thit k ph hp.

Vect ti t hp c bin np vo t bo ch thch ng, sau tin

hnh nui cy v tch chit plasmid ti t hp cha gen khng nguyn.

Khi s dng lm vaccine, ngi ta chpha long plasmid ti t hp c

cha gen khng nguyn v tim vo c th.


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3.2. NHM VACCINE C NGUN KHNG NGUYN KHNGC NHN LN

3.2.1. Vaccine v hot

Vaccine v hot (vaccine cht) l ch phm cha chnh tc nhn gy

bnh nhng b v hot (git cht) bng cc phng php khc nhau,

ch yu l l hc v ho hc.

3.2.2. Vaccine cha protein khng nguyn tinh ch

Mt s vi sinh vt trong qu trnh nhn ln v gy bnh c thi ra mi

trng mt s thnh phn protein c th thu nhn lm khng nguyn

ca vaccine.

mun s dng lm vaccine cn thit phi c nhng cht b tr thch hp

nu khng cc loi enzyme thu phn protein s d dng ph hu trc

khi gy kch thch min dch.


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3.2.3. Vaccine cha khng nguyn l protein ti t hp

Thnh phn vaccine chnh l protein tinh ch c sn xut bng cng ngh gen.

Gen khng nguyn ca i tng c ct ghp vo h thng vect thch hp i

vi mt loi vi sinh vt dng trong cng ngh gen.

Khi nui cy chng vi sinh vt ti t hp trong mi trng thch ng, protein

khng nguyn c tng hp vi lng ln. Sau tin hnh tch chit v tinh

sch lm vaccine.

3.3.4. Vaccine tng hp

Vaccine tng hp (synthetic vaccine) l loi ch cha duy nht mt on ngn

polypeptid khng nguyn (nhm quyt nh khng nguyn - epitope).

Sau khi c tng hp, mun lm vaccine chng phi c gn vo cc gi

c bn cht l cc ht polymer c kh nng hp ph cao.

Ngoi ra, chng cn c s dng vi nhng loi cht b tr tt khc.


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4. C S CNG NGH SN XUT VACCINE TH H MI

4.1. KHI NIM

Hn ch trong sn xut vaccine bng cng ngh c truyn? Khng phi tt c cc tc nhn ly nhim u c th nui cy c, do vy

khng th sn xut vaccine cho mt s bnh. Sn xut virus ng vt v virus ngi i hi mi trng nui cy t bo rt

t tin. Sn lng thp v tc nui cy virus ng vt v ngi bng nui cy

thng chm nn gi thnh sn xut vaccine cn cao. Cn bo m an ton trong phng th nghim v ngi sn xut h khng

b nhim cc tc nhn gy bnh. Nhng m vaccine c th khng cht hoc gim c lc khng trong qu

trnh sn xut, do sinh vt c c a vo vaccine v lan bnh mtcch v tnh.

Cc chng gim c lc c th hi c, nn i hi phi kim tra tip m bo khng xy ra s hi c. Khng phi tt c cc bnh (nh AIDS) u c th phng c qua s dng

vaccine truyn thng. a s cc vaccine hin nay c thi gian s dng hn ch v thng i hi

bo qun lnh duy tr cng hiu. y l mt vn kh cho cc nc cvng nng thn rng ln, khng c in.


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Phng php khc phc?

Cc gen c c th c loi b khi tc nhn ly nhim m vn cn kh nng kch

thch min dch. C th to c cc h thng chuyn giao sng khng gy bnh mang quyt nh

khng nguyn ring bit ca tc nhn gy bnh khng lin quan. dng ny, hthng chuyn giao to iu kin cm ng p ng min dch mnh trc tipchng li tc nhn gy bnh.

i vi cc tc nhn ly nhim khng th duy tr bng nui cy, cc gen quy nh

protein khng nguyn c th c phn lp, nhn dng v biu hin h thngvt ch khc. Cc protein ti t hp ny c th dng to vaccine.

C mt s tc nhn ly nhim khng trc tip ph hu tbo vt ch, thay vo biu hin bnh xy ra khi h min dch ca vt ch tn cng tbo (b ly nhim)ca chnh c th mnh. i vi cc bnh ny, c th to h thng dit tbo chc hiu, ch tn cng vo tbo b ly nhim loi b ngun p ng min dchkhng mong mun. Khi , gen quy nh mt protein dung hp c thit k. utin, mt phn ca protein dung hp s lin kt vi tbo b ly nhim. Sau ,phn cn li tiu dit tbo b ly nhim.


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Khi nim vaccine th h mi?

Vaccine th h mi (vaccine bng cng ngh gen) l cc ch phm cdng lm vaccine, gy min dch cho ngi v ng vt c to ra v sn

xut thng qua cc thao tc v k thut gen.

Cc loi vaccine th h mi?Vaccine xo gen

Vaccine phn t hay vaccine di nhm

Vaccine ti t hp c vect dn truyn

Vaccine DNA


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4.2. VACCINE XO GEN

Vaccine xo gen hay cn gi vaccine sng gim c lc sn xut bng k

thut di truyn.

Qu trnh sn xut c tin hnh bng vic:

- tch genome ra khi v capsid.

- Dng enzyme gii hn loi b cc gen c.

- To virus khng cn gen c lm vaccine a vo c th. Virus s nhn

ln trong tbo gy p ng min dch qua trung gian tbo.


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4.3. VACCINE PHN T

4.3.1. Vaccine phn t phng chng vim gan B

Gen m ho cho protein v virus HBV gm 3 gen:

gen pre-S1 m ho chui peptid c 128 amino acid, gen pre-S2 m ho chuipeptid c 55 amino acid v gen S m ho chui peptid c 226 amino acid


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Bng k thut PCR v cc k thut phn t khc, tch dng gen khng

nguyn S, gen m ho cho protein pre-S2, ti t hp v bin np vo t bo

nm men S. cerevisidae. Hoc c th tin hnh ring phc hp gen S vi pre-

S1 hoc phc hp gen S vi pre-S2, sau tin hnh biu hin trong h

thng biu hin v tinh sch protein ti t hp lm vaccine c hiu qu khc

nhau.

Gn y loi vaccine vim gan B mi c to nn t s biu hin c gen

khng nguyn: S, pre-S1 v pre-S2 ca HBV trong t bo trng chut t

vng Trung Quc. Vaccine ny c kh nng to phn ng min dch nhanh,

nng khng HBsAg cao min dch tt, to min dch chng bnh vim gan

lu di, lm gim t l mc bnh, gim t l ung th gan.


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4.3.2. Vaccine tng hp (vaccine peptid)

Hnh 3.1. Cc epitop bn ngoi c th gy min dch

Bn ngoi

Bn trong

Mng

1

2

4

5

3


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Ngi ta tng hp ho hc mt s on peptid ca khng nguyn ho tan VP1 ca

virus gy bnh l mm long mng (FMDV). Cc on peptid c tng hp gm: 141-

160, 151-160 v 200-213 nm u C ca VP1 v 9-24, 17-32, 25-41 nm gn u N

ca VP1 c lin kt vi protein mang tr.

Protein mangCu ni

Cc Peptid

ngn

Hnh 3.2. Cu trc ca vaccine peptid vi protein mang


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Cc on peptid ny c th nghim trn chut lang. Kt qu cho thy tim chng

mt ln peptid 141-160 gy khng th bo v ng vt chng c FMDV

nhng ln gp sau. Ngc li, khi tim VP1 hon chnh hoc cc peptid nm gn u N

cho nng khng th trung ho thp.

hiu qu, mt epitop phi gm mt on cc amino acid k tip nhau, m

iu ny thng khng xut hin t nhin.

Peptid phi c cu hnh ging nh epitop trn ht virus nguyn vn.

Mt epitop n c th khng gy min dch.

vaccine peptid c cc hn ch nht nh:


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4.3. VACCINE TI T HP C VECT DN TRUYN

4.3.1. c im chung* V nguyn l:

Loi vi sinh vt lm vect dn truyn phi l loi thng dng cho nhiu

loi v phi gim c hoc v c bng cch ct b hoc xo i mt hay

nhiu gen bng k thut gen. ng thi mun tr thnh vaccine, loi vi

sinh vt lm vect phi c ghp h thng biu hin gm promoter

mnh v mt hay vi gen khng nguyn m chng ta cn lm

vaccine.


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- i tng lm nhim v vect dn truyn: l cc loi virus v vi khun b ct b

nhng gen gy c v tnh gy bnh c gim n mt mc no m i tng

hng vaccine chu ng c, nhng bn thn chng vn duy tr c chu k sng

ca mnh trong c th. C nhiu i tng vi sinh vt c th s dng lm vect dn

truyn nh vi khun Salmonella, vi khun Lao, E. co li,... hoc mt s loi virus nh

virus u, virus adeno,...

* V mt cu trc: Cn phn bit 2 thnh phn chnh cu trc nnvaccine.

- i tng c ti t hp vo: l mt h thng c ghp vo vect c

cu trc gm 2 b phn. Mt l gen khng nguyn c tch t mt loi visinh vt gy bnh hay tc nhn gy bnh no . Mt b phn khc l

promoter c gn trc gen. Gen khng nguyn s chu s iu ho ca

vng promoter to ra protein khng nguyn kch thch c th sinh min

dch.


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4.3.2. Vaccine ti t hp vect dn truyn s dng vi khunSalmonella

u th ca vaccine c vect dn truyn Salmonella l d dng thao tc, d s dng, dnhn ln v d phn phi khng nguyn cho c th. Vic nui cy chng vi khun ti t

hp khng phc tp do m bo vic tip ging, nhn ging v sn xut vaccine

ng dng. Salmonella hin ang c s dng lm vect dn truyn cc loi gen

khng nguyn ca virus cm, HBV, virus st xut huyt, vi khun gy bnh ng rut,

vi khun t, lin cu khun, k sinh trng st rt,... Qu trnh to vaccine bt u bng vic to chng Salmonella nhc c. Bng k

thut gen ct b hoc xo hot tnh nhng gen gy bnh ca vi khun S.

typhimurium nh gen aroA, gen cya, gen crp hoc gen thyA,... nhng vn gi nguyn

cc c tnh sinh hc, sinh trng v pht trin nh vi khun ban u. Sau t hp

gen khng nguyn cng vi h thng khi ng c thit kkm theo c ghp vochng Salmonella nhc c. Chng ny tip nhn gen khng nguyn to thnh chng

vi khun ti t hp. Chng vi khun ti t hp c nui cy v to vaccine.


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4.3.3. Vaccine ti t hp c vect dn truyn s dng virus u

Hnh 3.3. S to vaccine ti t hp c vect dn truyn s dng virus u


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u nhc im ca vaccine ti t hp c vect dn truyn s dng

virus u:

virus u c kh nng kch thch c 3 loi hnh min dch gm min dch

dch th, min dch qua trung gian tbo v min dch nim mc. Min

dch c virus u tham gia thng ko di, bn lu, hot ph min dch

rng, thch ng nhiu loi, phng php vaccine n gin v tng i an

ton, gi thnh vaccine khng cao.

Tuy nhin, mi lo ngi ln nht khi s dng l mc d nhc c nhng

vn cn sng nn c kh nng trao i gen trong t nhin tr ln c

v c th ti gy bnh u ma.


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4.4. VACCINE DNA

4.4.1. Khi nim

Vaccine DNA l mt loi hnh vaccine c bit thuc th h mi nht,

chnh l mt plasmid vect biu hin c ti t hp gen khng nguyn

(hoc gene khng th) vi h thng promoter mnh c to ra bng cng

ngh gen.

Vaccine DNA gm 2 loi ch yu: loi vaccine tp hp cc gen khng nguyn

v loi vaccine tp hp cc gen khng th.

Gen khng nguyn c tch dng t virus, vi khun gy bnh. Trong t bo,

cc gen khng nguyn c phin m v dch m to nn cc protein khng

nguyn kch thch t bo sn xut khng th c hiu chng cc tc nhn gy

bnh. Vaccine DNA l tp hp cc gen m ho khng th c hiu thng

c ci gn vo cc vect biu hin mnh, khi a vo t bo cc gen c

dch m to cc khng th c hiu, gip t bo chng li s xm nhim ca

cc tc nhn gy bnh.


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4.4.2. u im ca vaccine DNA

Kh nng biu th ca gen khng nguyn rt mnh, thi gian sn xut protein

khng nguyn lu, do ngun khng nguyn duy tr trong thi gian di v cho

min dch tt ngay c vi c th thiu nng min dch.

vaccine DNA c kh nng kch thch loi hnh min dch qua trung gian tbo. u

th an ton ca vaccine DNA hn hn so vi vaccine ti t hp c vect dn

truyn ch: khng c hin tng trao i gen li c,

Vaccine DNA cc k bn vi nhit , khng i hi iu kin lnh nh cc loi

vaccine khc. Trong iu kin -20oC, vaccine DNA c th bo qun c hng nm

m vn gi c hot tnh, cn trong iu kin nhit thng c th gi c

trong 1-2 thng.

vaccine DNA l d dng to c vaccine a chng v a gi gip c th chng li

nhiu loi bnh nhim trng vi gi thnh thp.


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4.4.3. Phng thc s dng vaccine DNA

Phng php tim (phng php tim vo da v c)

Tim vo c

Tim ni b

Tim tnh mchTim di da

Tim bng

Phng php bn gene (phng php bn xuyn da v c)

Phng php a qua ng nim mc (nh mt, mi, ung)

Hiu qu p ng min dch ca cc phng php nh th no?


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4.4.4. V d v cng ngh sn xut vaccine DNA phng chng bnhGumboro

Thit k vect ti t hp c th s dng ring r hoc kt hp cc on gen m

ho khng nguyn VP2, VP3, VP4 to nn cc loi vaccine DNA c hiu qu

chng bnh khc nhau.

Vaccine DNA cha hai on gen m ho protein v virus gm on gen VP2 v

phn on A mang c 3 gen (VP2, VP3, VP4) tch dng t cc chng virus

Gumboro P40 v D78 s dng phng chng bnh cho gia cm Hungary c hiu

qu cao.

Vaccine DNA cha phn on A t chng Gumboro STC s dng trong phngchng virus Gumboro thnh cng ti M v nhiu nc khc v hin c ng

k bn quyn chnh thc ti M.


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Quy trnh cng ngh sn xut vaccine DNA chng bnh Gumboroc th thc hin theo cc bc c bn sau:

Tch chit RNA tng s t mu virus Gumboro

Thc hin phn ng RT-PCR vi cc cp mi c hiu thu nhn on

gen m ho protein khng nguyn cn thit.

To vect ti t hp gia on gen khng nguyn v vect tch dng

Thc hin k thut bin np, sng lc cc t bo mang vect ti t hp

Nui cy cc t bo sau bin np, thu nhn v kim tra DNA ti t hp

Th hiu lc vaccine DNA.


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VACCINE N C (VACCINE THC VT)


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Th no l vaccine n c?


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c im ca vaccine n c?


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Lm th no sn xut vaccine n c?


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CHNG 3

CNG NGH SN XUT ETHANOL SINH HC

1. XU THPHT TRIN CNG NGHIP ETHANOL SINH HC

2. C S HO SINH CA NGNH CNG NGHIP SN XUT ETHANOL

SINH HC

3. CNG NGH SN XUT ETHANOL T NGUN NGUYN LIU NG,

TINH BT

4. CNG NGH SN XUT ETHANOL T NGUYN LIU LIGNOCELLULOSE


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Trung Quc l quc gia sn xut v s dng ethanol nhin liu ln th 3

sau Braxin v M. Nm 2004 a vo hot ng nh my sn xut

ethanol ln nht th gii vi cng sut 600 000 tn/nm, tng sn lng

ethanol c nc trn 3,5 triu m3. T thng 6/2002, nc ny quyt

nh s dng xng pha 10% cn khan 5 thnh ph v n cui nm 2006

tng thm 27 thnh ph ng dn khc.

n s dng xng pha 5% cn 9 bang v 4 tiu vng t ngy

1/1/2003.

Thi Lan cng l mt trong nhng quc gia tch cc u t vo nhin liu

sinh hc. Nm 2004, nc ny sn xut trn 280 000 m3 v u t

thm 20 nh my n nm 2015 c trn 2,5 triu m3 ethanol lm nhin

liu.


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2. C S HO SINH CA NGNH CNG NGHIP SN XUTETHANOL SINH HC


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Hnh 3.2. Con ng ng phn


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2.2. QU TRNH LN MEN


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3. CNG NGH SN XUT ETHANOL T NGUN NGUYN LIUNG, TINH BT

Loi nguynliu

Nc(%)

Protein(%)

Chtbo (%)

Gluxit

(%)

Celluose(%)

Tro(%)

Sn ti 70,25 1,102 0,41 26,58 1,11 0,54

Sn kh 13,12 0,205 0,41 74,74 1,11 1,69

Khoai ty ti 74,9 1,99 0,15 20,8 0,98 1,09

Khoai lang ti 68,1 1,6 0,5 27,9 0,9 1,0Khoai lang kh 12,9 6,1 0,5 76,7 1,4 2,4

Ng 14 10 4,6 67,9 2,2 1,3

CcCc loiloiTBTB cc ss dngdng


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HiHi thth hh th lth l tt tt l il i


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HiuHiu ququthuthu nhnnhn ethanolethanoltt mtmt ss loiloinguynnguyn liuliu??


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SS tngtngqutqut cngcng nghngh snsn xutxutEthanolEthanol


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a. Nghin nguyn liu

Mc ch: Ph vcu trc mng t bo thc vt, to iu kin gii phng ccht tinh bt khi cc m, khi a vo nu nhit v p sut ph hpbin tinh bt thnh dng ha tan.

C2phng php nghin nguyn liu:

- Nghin t: nguyn liu c nhng vo nc hay axit ho tan tch thnhcc thnh phn trc khi chuyn ho tinh bt thnh ng ln men thnh

ethanol.- Nghin kh: nguyn liu c nghin thnh bt mn v ch bin m khngphn tch thnh cc thnh phn. Nguyn liu dng ht c a vo mynghin ba ( Vit Nam).

+ Nghin th: ba c chiu dy 6-10mm.

+ Nghin nh: ba c chiu dy 2-3mm.+ Tc quay ca my nghin: 2750 vng/ pht.


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b. Nu nguyn liu

- Mc ch: ph vcu trc dng ht ca tinh bt to thnh

cc mch pectin v amilopectin nhit cao to iu

kin thun li cho qu trnh ng ha.

- C 3 phng php nu nguyn liu:

Nu gin on

Nu bn lin tc

Nu lin tc


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Nu gin on: Ton b qu trnh nu c thchin trong cng mt ni.

- Ccbc: Cho nguyn liu vo ni, cho ton b

nc vo ni vi t l 3,5 4lt/kg nguyn liutu thuc vo hm lng tinh bt, y np kncho my khuy lm vic ri bt u xng hi saocho 45 60 pht th p sut trong ni t yucu. i vi nguyn liu nghin th p sutt yu cu khong 3 3,5 kg/cm2.

- Thi gian nu t 60 70 pht c tnh t khi

t p sut yu cu, nhit tngng l 135 1400C.

- Khi nu c th cho thm H2SO4 long vi hmlng 2 4 kg/tn nguyn liu s nu nhanhchn hn v dch cho tinh bt t hoc khngblo ho. Tuy nhin nu cho nhiu s khng c lido n mn thit b v lm gim hot tnh ca

amylaza.

Nu bn lin tc : Qu trnh nu c tin


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S h thng nu bn lin tc

hnh trong 3 ni khc nhau v chia thnh nus b, nu chn v nu chn thm. Phngphp ny tit kim c hit v cho nngsut cao ng thi thi gian nu c rt ngn

tuy nhinphi sdng ti 3 ni nu do tn km cho thitb.

- Nu sbc thc hin trong thit b hnhtr. Lc u cho lng nc 40 500C vocng vi t l 3,5 4lt/kg bt khuy trn ridng hi th t ni nu chn thm undung dch bt ti 7 0 8 50C v duy tr khong

50 60 pht sau a sang ni nu chn.- Nu chn: p sut nu 2,8 3,2 kg/cm2, nhit

130 1350C, thi gian nu khong 60pht.

- Dch cho sau khi nu chn c a sang ninu chn thm p sut nu t 0,5 0,7 kg/cm2,nhit 105 1060C, thi gian nu 50 60pht.

- Nu lin tc : Nu lin tc cho php c cnng sut cao, tn hao nng lng t v thigian nu c rt ngn. Tuy nhin phngphp ny i hi cc iu kin nghim ngt

nh v kch thc ca bt phi c mt t lxc nh,vic cung cp in, hi nc,ncphi n nh. Qu trnh nu lin tc cng cthc hin thng qua 3 giai on thc hintrong 3 thitb khc nhau l : Nu sb, nuchn v nu chn thm.

Q h hQ h h


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Qu trnh ng ho dch cho nuc th thc hin gin on hay lintc. Tuy nhin v c bn th n baogm cc cng on sau:

- Lm lnh dch cho ti nhit ng ho.

- Cho ch phm amylaza vo dchcho v gi nhit trn trongthi gian xc nh amylazachuyn ho tinh bt thnh dng.

- Lm lnh dch ng ho ti nhit ln men.

- Sau ng ho nhit dch ngl 300C hm lng ng t 80 100g/l.

c. Qu trnh ng hac. Qu trnh ng ha

d Qu trnh ln mend Qu trnh ln men


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d. Qu trnh ln mend. Qu trnh ln men

CChng nm men c s dnghng nm men c s dng

N N


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Nm mcNm mc


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Vi khunVi khun

Mt s vi khun:

- Sasina ventriculi,

Zymononas mobylis.

- Ngoi ra cn c vi khunlactic d hnh, vi khun

ng rut hay Clostridium


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Ln men ru l mt qu trnh ln men ym khdin ra rt

phc tp, bao gm cc qu trnh sinh ha hc v cc qu trnhvi sinh vt.

Qu trnh ln men c th c thc hin gin on hay lintc :

- Ln men gin on: Men ging v dch ng cng cbm vo thng, lng men ging chim khong 10% th tchthng ln men, dch ng c bm t t vo thng(khong 68 gi/1 thng) hn ch s pht trin ca tpkhun. Thi gian ln men l 3 ngy. Ln men gin on cu im l d lm,khi nhim tp khun d s l tuy nhinnng sut thu c thp.

- Ln men lin tc : Dch ng v men


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c cho vo thng u gi l thngln men chnh,lun cha mt lng lnt bo trong 1ml dch. Khi y thngu th dch ln men s chy tip theosang cc thng bn cnh v cui cngl thng cha gim chn.

+ Qu trnh ln men c tin hnh nhsau: u tin men ging c phttrin ri c a vo thng ln mencp I. Sau c a sang thng lnmen cp II ri mi c a sangthng ln men chnh, sau c asang thng ln men ph.

+ Vn ch yu ca ln men lin tc lphi lun lun khng ch s t bo thng ln men chnh khong 100 120triu TB / ml v phi m bo vtrng.

+ u im: thng ln men chnh nn ln men xy ranhanh,hn ch c pht trin ca tp khun. Mtkhc do chua ca dch ng cao, pH thp cng lyu t khng thun li cho vi khun v nm menhoang di. Sau 24 gi c 78% ng c ln

men, trong khi ln men gin on mi ch t42%.

+ Nhit ln men: i vi thng ln men chnh nngi 25 270C thng ln men ph t 27 300C ccthng cn li 27 280C. Qu trnh ln men s ktthc sau 60 62 giso vi ln men gin on l 72gi. Sau qu trnh ln men nhit dm 34350C .


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e. Qu trnh chng cte. Qu trnh chng ct

Khi qu trnh ln men kt thc, ta tin hnh chng ct thuc ru thnh phm.

Qu trnh chng ct ru nhm tch hn hp ru v nc

c nhit si khc nhau. p sut thng, ru si v bchi 780C, ca nc l 1000C...

Cc hnh thc chng ct:

- Chng ct lin tc.

- Chng ct gin on.- Chng ct bn lin tc.


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MtMt ss phngphng phpphp lmlm tngtng cncn

dng cc cht ht m (nh vi sng, photphat kali, sunfat ng).

phng php chng ct thng, chng chn khng p sut thp (0,0525

atm)

Phng php chng luyn ng ph: y l phng php cho thm cu t

th ba (benzen, clorofooc, toluen...) vo nhm thay i bay hi tng i

ca hai cu t trong h ban u. C ngha l thm cu t th ba vo, n to

thnh vi cu t d bay hi (hay vi c hai cu t) dung dch ng ph c

bay hi ln hn v sn phm y thp l cu t dng nguyn cht.

Phng php hp ph trn ry phn t


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4. CNG NGH SN XUT ETHANOL T NGUYN LIU


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LIGNOCELLULOSE

Nguyn liu Lignocellulose


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Cc giai on sn xutethanol

4 1 THU PHN BNG ACID C


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4.1. THU PHN BNG ACID C

Cng ngh ny da trn vic s dng axits c ph hu cu trc tinh th cacellulose sau thu phn bng axit long. Cc khu tch axit khi ng, thu

hi v c c axit l nhng cng on mu cht ca qui trnh.

Trong cng ngh ny, axit c s ph hu cc lin kt hydro gia cc mch

cellulose v chuyn chng sang trng thi v nh hnh. Sau , cellulose s

hnh thnh trng thi gelatin vi axit v tr ln mn cm vi phn ng t thu

phn. Khi pha long vi nc v di tc ng ca nhit, cellulose nhanh

chng b thu phn thnh glucose.


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Hnh 3:S h thng sn xutethanolbngcngngh thyphn dngaxitc(Arkenol - HoaK)

T i t h i h khi th t h 10% l b it


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Trong qui trnh ny, sinh khi thc vt c hm m 10% c x l bng axit

sunfuric nng 70 77%. Lng axit s dng so vi sinh khi l 1,25:1 v nhit

c gi mc di 50oC. Sau nc c b sung pha long axit ti

nng 20 30% v gia nhit ti 100oC, gi trong thi gian 1h. Dch dng gel

c ly ra khi b phn ng v lc p ly dch ng v axit. phn cht rn

cn li s c ti thu phn ln hai. Dch ng v axit s c phn tch s

dng ct sc k. Cng on ln men tip theo c th chuyn ho c ng

thnh ethanol vi hiu sut 85% vi ng xylose v 92% vi glucose. Hng

Arkenol tuyn b c th thu hi 97% axit trong qui trnh ca mnh.

4.2. THU PHN BNG ACID LONG


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Hnh 4.2. S h thng sn xut ethanol bng cng ngh thu phn dngaxit long

h h h h l l b d d h


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Trong giai on thu phn th nht, nguyn liu c x l bng dung dch axit

sunfuric 0,7% nhit 190oC vi thi gian 3 pht.

Giai on 2: c thc hin 215oC vi nng axit l 0,4% v thi gian l 3 pht.

Dch thy phn t mi qu trnh c thu hi, trung ho v ln men to ethanol.

Cellulose v lignin cn li dng rn c s dng lm nhin liu cho sn xut hi.

4.3. THU PHN BNG ENZYME


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4.3. THU PHN BNG ENZYME

H enzyme tham gia v c ch thy phn cellulose

Quy trnh sn xut:


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Ct/nghin nh X l s b bngaxit long

Sn xutenzyme

Ln men

Thy phncellulose bng

enzymeX l bthi rn

Thu hiethanol

Dch thy phn

cellulose(ng glucose)

Dch thy phn

hemicellulose(ng xylose)

y

X l s b nguyn liu bng axit, kim, nhit nhm m bo khnng tip cn ca enzyme ti cc lin kt ca phn t cellulose.

Cng ngh SHF: thy phn v ln men ring bit.

S tmttquy trnh cngngh SHF

Cng ngh SSF: thy phn v ln men ng thi.


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Cng ngh SSF: thy phn v ln men ng thi.

Gim s lng thit b: b qua cng on thy phn ring bit.

Gim c ch ngc ca sn phm vi enzyme: glucose trong dung dch sc ch hot ng thy phn cellubiose ca -glucosidase.

Hiu sut chuyn ha cellulose thnh ethanol tng 40% (so vi SHF).

Quy trnh ci bin SSCF: ln men ng thi nhiu loi ng.

Ct/nghin nhX l s bbng axit

long

ng thing hav ln men

Sn xutenzyme

X l bthi rn

Thu hiethanol

S tmttquytrnhSSCF

Nhng kh khn trong quy trnh cng ngh?


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Nhng kh khn trong quy trnh cng ngh?

SSkhckhc nhaunhau trongtrong

nhitnhit titi uuchocho

hothot ngngenzymeenzyme cellulasecellulase

((4545--5050ooC)C) vv nhitnhit thchthch hphp chocho vivi sinhsinh vtvt lnln menmen ethanolethanol ((2828--

3535ooC)C)..

MtMt ss thnhthnh phnphn dchdch thythy phnphn,, ethanolethanol cngcng cc tctc ngng cc chch

enzymeenzyme cellulasecellulase.. GiGi thnhthnh cellulasecellulase ququ caocao

Nhng phng hng khc phc?


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Bai Giang Cong Nghe Hoa Sinh 2010