Cong ngheSINH HOC bllDfC

• •(DUNG CHO DAO TAO DlfQC Si DAI HOC)

Chu bien: GS. TS. NGUYEN VAN THANH

Khong khi vao

Khong khi ra

Loc vo trung

Binh chura moi tri/dng

Ong dan tran

Noi len men

Binh thu hoach

CDNHA XUAT BAN GIAO DUC VIET NAM

sv __BO YTE

■

CONG NGHE SINH HOC Dime■ ■

(DUNG CHO DAO TAO Dl/OC SI DAI HOC)MA SO: D.20.Z.09

NHA XUAT BAN GlAO DUC VIET NAMHA N Q I-2 0 0 9

Chi dao bien soanVU KHOA HOC VA DAO TAO - B0 Y TE

Chu bien

GS.TS. NGUYEN VAN THANH

Nhufng ngutoi bien soan

GS.TS. NGUYEN VAN THANH TS. TRAN CAT DONG PGS.TS. TRAN THU HOA PGS.TS. BUI TUNG HIEP TS. NGUYEN TRONG HIEP ThS. HUYNH THI NGOC LAN

Tham gia to chtfc ban thao

ThS. PHI VAN THAM TS. NGUYEN MANH PHA

© Ban quyen thuoc Bo Y te (Vu Khoa hoc va Dao tao)

1040 - 2009/CXB/5 - 1960/GD Ma so : 7K829Y9 - DAI

LCJI Gldl THIEU

Thuc hien mot so dieu cua Luat Giao due, Bo Giao due & Dao tao va Bo Y te da ban hanh chtfdng trinh khung dao tao DUdc si dai hoc. Bo Y te to chiic bien soan tai lieu day - hoc cac mon cO sd va chuyen mon theo chUOng trinh tren nh&m tting budc xay dung bo sach dat chuan chuyen mon trong cong tac dao tao nhan lUc y te.

Cuo'n Cong nghe sinh hpc DtfOc dUOc bien soan dUa vao chUOng trinh giao due cua khoa DUOc Dai hoc Y-DUcJc Tp. Ho Chi Minh tren cO sd chUOng trinh khung da dUdc phe duyet. Sach dUOc cac giao sU, tien si, cac nha giao co kinh nghiem cua Bo mon Vi sinh - ky sinh bien soan theo phUdng cham: kien thilc cd ban, he thong; noi dung chinh xac, khoa hoc; cap nhat cac tien bo khoa hoc, ky thuat hien dai va thUc tien Viet Nam.

Cuon Cong nghe sinh hoc DifOc da diidc Hoi dong chuyen mon tham dinh sach va tai lieu day - hoc chuyen nganh Dude si dai hoc cua Bo Y te tham dinh nam 2009. Bo Y te quyet dinh ban hanh la tai lieu day — hoc dat chuan chuyen mon cua Nganh trong giai doan hien nay. Trong thdi gian tii 3 den 5 nam, sach phai dUdc chinh ly, bo sung va cap nhat.

Bo Y te chan thanh cam dn cac tac gia va Hoi dong chuyen mon tham dinh da giup hoan thanh cuoh sach; cam dn GS.TSKH. Nguyen Van Dip va PGS.TS. Cao Van Thu da doc va phan bien de cuon sach sdm hoan thanh, kip thdi phuc vu cho cong tac dao tao nhan liic y te.

Lan dau xuat ban, chung toi mong nhan dUdc y kien dong gop cua dong nghiep, cac ban sinh vien va cac doc gia de lan xuat ban sau sach dUdc hoan thien hdn.

VU KHOA HOC VA DAO TAO - B0 Y TE

3

LOI NOI DAU

Cong nghe sinh hoc da xuat hien tii khi loai ngUdi con chUa co hieu biet day du ve cac he thong song. Cac cong nghe sd khai nay dUdc hinh thanh nhd kinh nghiem va chu yeu lien quan den viec che bien thUc pham va do uong len men. Den the ky XIX, khi Louis Pasteur dat nen mong cho vi sinh hoc thuc nghiem, cong nghe sinh hoc budc vao thdi ky mdi dUa tren khoa hoc sinh hoc. Tti luc do, cong nghe len men hien dai da dUdc phat trien de san xuat cac chat chuyen hoa tu: vi sinh vat nham phuc vu ddi song va cac nganh cong nghiep khac. Cung trong thdi ky nay, vaccin da dUdc phat minh va day co the coi la san pham cong nghe sinh hoc dUdc dau tien, nhiing mot nganh cong nghiep sinh hoc dUdc thUc sU chUa dUdc hinh thanh.

Su ra ddi cua cong nghiep sinh hoc dUdc thuc sU dUdc danh dau bang viec san xuat penicillin va streptomycin nhd cong nghe len men vao nhiJng nam 1940. Den cuoi nhQng nam 1970, cong nghe sinh hoc dUdc da budc vao mot giai doan mdi vdi viec san xuat insulin ngUdi bang ky thuat tai to hdp di truyen. Chinh viec ap dung manh me cong nghe gen trong san xuat cac protein tri lieu da lam cho cong nghe sinh hoc dUdc trd thanh dong luc chinh de dUa cong nghe nay vao giai doan hien dai vdi sU hoi tu cua cong nghe gen va cong nghe len men. Ngay nay, cong nghe sinh hoc dUdc van dUa tren nen tang cong nghe len men de san xuat ra san pham, nhUng viec phat trien cac san pham mdi chu yeu dUa vao cong nghe gen, enzym va te bao. Mat khac, cac he thong san xuat khong con la te bao vi sinh vat nCia ma co the la te bao dong vat, thUc vat, sinh vat chuyen gen va tham chi cac he thong phi sinh vat.

Cong nghe sinh hoc dvfdc ngay nay dong vai trd quan trong doi vdi nganh DUdc. Trong cac nam gan day, thuoc cong nghe sinh hoc chiem da so cac thuoc mdi dUdc phat trien va cap phep. Cac hang dUdc pham ldn tren the gidi deu co bo phan cong nghe sinh hoc rat manh hoac sap nhap vdi cac hang cong nghe sinh hoc. Cong nghe sinh hoc trong nganh Y-DUdc chiem den 80-90% tong doanh thu cua cong nghe sinh hoc noi chung. DUdc pham cong nghe sinh hoc hien nay khong con la cac san pham chuyen hoa vi sinh vat ma chu yeu la cac protein tai to hdp, acid nucleic va te bao.

Trong sach nay, chung toi co’ gang khai quat cac cong nghe nen tang cua cong nghe sinh hoc dUdc va cung cap cac thi du ve cong nghe san xuat cua mot so" dUdc pham cu the. Sach dUdc chia lam nam chUdng tUdng ilng vdi cac cong nghe nen do la: cong nghe len men, cong nghe enzym, cong nghe te bao, cong nghe gen va cong nghe mien dich, trong moi chUdng deu co khai quat cae van de chung cua cong nghe va ting dung san xuat mot so" san pham cu the.

Sach dUdc xuat ban lan dau do do khong tranh khoi thiet sot, rat mong nhan dUdc sU dong gop y kien cua ban doc.

cA c t Ac GIA

5

M TJC L U C

Ldi gidi thieu................................................................................................................3Ldi noi dau................................................... ...............................................................5

Bdi 1 Md dau.............................................. .................................................................91 . Khai niem................................................................................................................92. Sii phat trien cua Cong nghe sinh hoc the gidi.................................................... 133. Su phat trien cua Cong nghe sinh hoc Viet Nam................................................ 144. Phat trien Cong nghe sinh hoc Viet Nam den nam 2020................................... . 16

Chu’dng 1 . CONG NGHE LEN MEN......................................................................... 20Bdi 2. Khai niem va phtfOng phap len men cong nghiep.................................. 20

1 . Dai cifdng...............................................................................................................202. Chung vi sinh vat................................................................................................. 213. Moi trifdng len men.............................................................................................. 364. He thong len men................................................................................................. 465. Giam sat qua trinh len men................................................................................. 58

Bdi 3. San xuat khang sinh..................................................................................... 611 . Md dau...................................................................................................................612. San xuat benzylpenicillin............................................................................ ........623. San xuat penicillin V ........................................................................................... 684. San xuat cephalosporin C .....................................................................................685. San xuat erythromycin.........................................................................................72

Bdi 4. Thi^c pham chufc nang...................................................................................811 . Khai niem......................................................................... ...................... ..............812. Prebiotic................................................................................................................ 833. Probiotic................................................................................................................ 854. Nhiing khuynh hiidng trong ttfdng lai................................................................100

Bdi 5. San xuat mot so san pham len men khac.................. *.............................1031 . San xuat acid hQu cd...........................................................................................1032. San xuat acid amin bang con diidng len men.....................................................1183. San xuat mot so" vitamin.....................................................................................132

ChtfOng II. CONG NGHE ENZYM...........................................................................145Bai 6. Khai quat ve cong nghe enzym - protein ................... ............................145

1 . Khai niem............................................................................................................ 1452. Xuc tac sinh hoc.................................................................................................. 1463. Nguon cung cap enzym....................................................................................... 1495. Enzym co dinh..................... ................................................................................158

Bdi 7. Ung dung enzym trong nganh DtfOc......................................................... 1701 . Enzym tri lieu..................................................................................................... 1702. San xuat thuoc bing cong nghe enzym........................................................ ......ig i

7

Chtfofng III. CONG NGHE TE BAO......................................................................... I91Bdi 8. Cong nghe nuoi cay te bao va ting dung trong nganh Y-D ifdc.......... 191

1 . Cong nghe nuoi cay te bao...................................................................................1912. tTng dung cong nghe nuoi cay te bao...................................................................198

Bdi 9. Cong nghe te bao goc....................................................................................2011. Khai niem............................................................................................................. 2012. Trien vong va kho khan trong viec ting dung te bao goc.................................... 2073. San xuat phoi IVF (in vitro fertility)...................................................................209

Chticfng IV. CONG NGHE GEN................................................................................211Bdi 10. Cong cu va ky thuat cd ban.......................................................................211

1. LUdc sti cong nghe gen......................................................................................... 2 1 12. Cong cu cd ban..................................................................................................... 2123. Ky thuat thao tac gen.......................................................................................... 2204. Toi Uu hoa sU bieu hien cua gen tai to hdp..........................................................225

Bdi 11. U”ng dung cong nghe gen ........................................................................... 2301 . San xuat protein tai to hdp.................................................................................. 2302. Lieu phap gen........................................................................................................249

Bdi 12. Cac phticfng phap chan doan phan tti.....................................................2551. Nhiing phtidng phap cd ban trong chan doan phan tti........................................2562. L/ng dung cua chan doan phan tiJt........................................................................ 272

ChtiOng V. CONG NGHE MIEN DICH..................................................................... 283Bdi 13. San xuat v a ccin ........................................................................................... 283

1. Md dau................................................................................................................... 2832. Phan loai vaccin....................................................................................................2843. PhUdng phap san xuat vaccin.............................................................................. 2864. Ta chat mien dich..................................................................................................293

Bdi 14. Huyet thanh va khang the.............................. ...........................................2971. Lich sti hinh thanh................................................................................................2972. Cac che pham gamma globulin............................................................................ 2983. Du phong bing globulin huyet thanh miln dich.................................................3014. Du phong bang globulin miln dich cao................................................................ 3025. Miln dich lieu phap vdi khang the ddn dong......................................................3036. Mien dich lieu phap trong dieu tri ung thii.........................................................303Dap an Tuf ltfdng gia..................................................................................................305Tai lieu tham khao va doc them.............................................................................. 306Muc luc tra ctiu (index).............................................................................................308

8

Bai 1

MCfDAU

MUC TIEU

1. Trinh bay ditdc cac khai niem ve cong nghe sinh hoc va cong nghe sinh hoc diCdc.2. Trinh bay ditdc qua trinh phat trien cua cong nghe sinh hoc the gidi va Viet Nam.3. Chi ra ditdc cac dac diem chinh cua cong nghe sinh hoc.

1. KHAI NIEM

1.1. Dinh nghla

Thuat ngti “cong nghe sinh hoc” lan dau tien diidc nhac den bdi Karl Ereky, mot ky sU ngUdi Hungary vao nam 1919. Luc bay gid, no nham chi kieu san xuat trong do san pham dUdc tao ra tti nguyen lieu dau vdi sU trd giup cua sinh vat song. Ereky da hinh dung ra mot “thdi dai hoa sinh” tUdng tu nhu “thdi do da” va “thdi do sat” .

Ngay nay, co nhieu dinh nghia ve cong nghe sinh hoc, tuy nhien dinh nghia cong nghe sinh hoc la “bat ky cong cu nao difdc sti dung de thao tac tren cac sinh vat, cac he thong sinh hoc hay mot phan cua no de tao ra san pham hay dich vu” dUdc cong nhan rong rai. Tti dinh nghia nay ta co the thay r&ng, ngay ca trUdc khi loai ngUdi hieu dtfdc sinh hoc la gi ho da s \i dung Cong nghe sinh hoc (CNSH) de san xuat rUdu va banh mi, da biet bien doi cac dac tlnh tU nhien cua sinh vat nham thu cac san pham co ldi. Vdi kinh nghiem va cac kien thiic tich luy diidc den nay CNSH da co tam anh hUdng sau rong den nhieu khia canh ddi song con ngvfdi, md ra nhieu trien vong ve nang suat nong nghiep hay dieu tri benh tat, cai thien chat ltfdng cuoc song.

Vdi viec tich luy kien thijfc va kinh nghiem vdi cac ky thuat sinh hoc hien dai, dinh nghla CNSH dUdc md rong bao gom cac ting dung cua cong nghe tai to hdp di truyen, cong nghe nuoi cay te bao trong san xuat va dich vu. CNSH hien dai va co dien khong khac nhau ve nguyen ly nhting khac nhau ve ky thuat dildc sti dung. Vi du, viec cai thien di truyen co dien va cac ky thuat di truyen phan tti co cung muc dich nham cai thien san pham de thu dtidc nhieu ldi ich hdn, tuy nhien, cac cai tien d mtic phan tti cho ket qua chac chan hdn. Hau het cac thuoc tinh cua san pham sinh hoc deu dtidc bien doi theo htidng co ldi bdi nha san xuat. Phtidng phap

9

co dien de bien doi mot tinh trang nao do can nhieu thdi gian va quan trong hdn no phu thuoc nhieu vao sd ton tai cua thuoc tinh do trong tu nhien. Vi du, de cai thien tinh de khang vdi sau benh, cac nha khoa hoc can tim ra nguon cung cap kieu gen de khang tUdng thich vdi loai can cai tao mdi co the lai dUdc. Doi khi dieu nay gan nhu khong the dUdc vi tinh trang do khong ton tai trong kho du trti gen cua loai hoac cac loai lan can. Vdi cong nghe gen, ngifdi ta co the dUa mot gen de khang nhu vay tti mot loai khong tUdng thich vao loai can cai thien vdi ket qua tin cay dUdc ma khong can quan tam den sU ttfdng thich ve bo may sinh san gitia hai loai. Dieu nay cho phep md rong gan nhu vo han kho dii trti gen.

Cong nghe sinh hoc hien dai dUdc thuc hien d mtic do phan til, nghia la cac gidi han triidc day co lien quan den loai deu khong con ntia. Do la vi tat ca cac loai deu chtia ADN, phan tti mang toan bo thong tinh di truyen vdi ma di truyen pho bien cho tat ca cac loai. Cac phan tti ADN nay co the diidc thao tac, xti ly, bien doi ben trong hay ben ngoai te bao, co the di chuyen tti te bao nay sang te bao khac, vUdt qua cac bien gidi cua loai sinh hoc.

1.2, Phan loai cong nghe sinh hoc

Theo sti phat trien cua ky thuat san xuat, CNSH dtidc dtidc chia lam ba the he:

- The he mot: san xuat cac san pham thuc pham va nufdc uong len men. Ky thuat diidc sti dung bao gom len men hd vdi giong tiJ nhien va phan 16n la chifa difdc biet ro, nuoi trong va nuoi cay mo thuc vat. Giong san xuat dtfdc cai tien thong qua chon loc tu nhien hay lai tao.

- The he hai: sti dung ky thuat nuoi cay te bao hay nuoi cay mo de san xuat. San pham cua the he nay gom khang sinh, enzym, vitamin, acid amin, acid htiu cd, dung moi. Ky thuat dUdc sti dung bao gom gay dot bien va chon loc chung vi sinh vat va phifdng phap len men kin, thuan chung, dUdc toi i*u hoa de sinh san pham toi da.

- The he ba chinh la CNSH hien dai. No bao gom cac ky thuat tai to hdp di truyen. Ung dung cong nghiep cua no bao gom diidc pham, nong nghiep, hoa chat, y hoc. Cac san pham hien nay dang la cac protein tri lieu, chan doan, vaccin the he hai, ba, cai tao giong nong nghiep va xti ly moi trUdng.

Can liiu y rang ngay nay tat ca cac the he CNSH deu sti dung cac ky thuat cua the he ba, bat ke ho sti dung cong nghe the he mot, hai hay ba de san xuat. Do do phat trien CNSH can dat trong tam phat trien cac ky thuat the he ba de tao tien de thtic day chung.

Xet ve pham vi ting dung CNSH co the dtitic chia la ba loai:- Cong nghe sinh hoc xanh (Green Biotechnology): CNSH ap dung trong

nong nghiep va xti ly moi trifdng.

10

- Cong nghe sinh hoc trang (White Biotechnology): sti dung xuc tac sinh hoc va cong nghe len men de tao ra cac san pham cong nghiep nhti hoa chat hay enzym.

- Cong nghe sinh hoc do (Red Biotechnology): CNSH ting dung trong Y—Dtidc. Chan doan va dieu tri benh. San xuat dUdc pham.

1.3. Ky nguyen cong nghe sinh hoc hien dai

Cong nghe sinh hoc hien dai vdi nhiing tiem nang ting dung to ldn da thu hut sU quan tam cua nhieu ngtidi, tuy nhien no cung lam mot so" ngtidi e ngai va tham chi phan doi. Hai dac diem chinh cua CNSH la nguyen nhan cua sti e ngai nay la toe do ap dung cua no trong nhieu llnh vUc kinh te trong cac nam gan day va cach tiep can dot ngot cua no vao thi trUdng tieu dung. Co the thay, vi du, thdi gian tti luc phat hien ra cong nghe den luc san pham co mat tren thi trtidng dUdc rut ngan rat nhieu so vdi 50 - 100 nam trUdc day. TrUdc day, mot sang che, nhu may truyen hinh, tti luc phoi thai phai mat 20 — 30 nam mdi pho bien. Nhting ngay nay, nhieu san pham CNSH, nhu thtic pham bien doi gen, da dtidc dtia ra sti dung tren thi trtidng ngay ca trtidc khi moi ngtidi cd dtidc nhan thtic day du ve no. Sti thieu nhan thtic cua cong chung da tao ra tam ly sd hai doi vdi cac san pham cua CNSH.

Cd the so sanh vdi thdi diem ma dong cd hdi ntidc va cac day chuyen cong nghiep dUdc phat minh. Khi do ngUdi ta sd hai trUdc sU cai thien nang suat ma cac phat minh nay dem lai.

Bang 1.1. So sanh thdi gian phat trien va thifdng mai hoa mot so cong nghe

Cong nghe Phat minh Bat dau san xuat Thdi gian phat trien

Viet may 1888 1938 50 nam

Truyen hinh 1907 1936 29 n£m

Giong cay trong bien doi gen 1983 1994 11 nSm

1.4. Cac dac diem cua cong nghe sinh hoc

1. La mot ky thuat da nang. Ung dung rong rai, va cd tinh chuyen doi qua lai gitia cac linh vtic, vi du mot ky thuat dtidc ap dung cho y te cung cd the dtidc sti dung cho nong nghiep.

2 . Ddi hoi miic do nghien ciiu cao. Ty le dau tti cho nghien ctiu de phat trien mot san pham mdi cao hdn cac nganh khac 5 —10 lan va chiem den 40 - 50% ldi ttic. Do do, sti lien he gitia cac cd sd han lam vdi cd sd ting dung san xuat phai het stic chat che.

11