Đề Cương Kháng Sinh

8/3/2019 Cng Khng Sinh

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Cu 1: nh ngha khng sinh, n v & p/loi k/sinh?1) nh ngha khng sinh:- C rt nhiu /ngha v k/sinh, song /ngha cht ch theo

Waksmann (1942) c ngha kinh in nh sau: K/sinh lnhng sp trao i cht t nhin c cc vsv to ra, c t/dng cch s ptr or tiu dit chn lc /vi cc vsv khc.Vd: k/sinh penicillin.- n nm 1972 c khong 4076 k/sinh c tm thy, ngynay s k/sinh mi p/lp c hng nm l khong 100.2) n v khng sinh:

* L lg k/sinh ti thiu ha tan trong 1 th tch mi trngx/nh c t/dng c ch hay tiu dit vsv.

* Mi k/sinh c /v h/tnh s/hc ring:+ /v hot lc of penicillin t nht h/tan vo 50ml canh

thang c t/dng c ch s ptr of Staphylococcus aureus 209p.+ /v hot lc of streptomycin l lg streptomycin t nht

h/tan trong 1ml canh thang c t/dng c ch s ptr of E.coli.3) P/loi k/sinh khng sinh:* P/loi k/sinh nhm khi qut 1 cch h/thng danh mc cck/sinh ngy cng nhiu thm. C th p/loi da vo p t/dng,c ch t/dng. p/loi theo ng/gc, con ng sinh tng hp hayc/trc ha hc. Nhng p/loi theo c/trc ha hc l khoa hcnht.

* Gii thiu p/loi theo c/trc ha hc & v d: c 9 nhmk/sinh chinh.- Cc k/sinh cacbonhydrat.Vd: cc saccarid thun nht (nojirimycin); cc glycolipid

(moenomycin), - Cc lacton macrocyclic

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Vd: cc k/sinh macrolid (erythromycin), cc anzamycin(rifamycin),- Cc k/sinh quinon & dn xut.

Vd: Cc tetracycline (tetracyclin), cc benzoquinon(mitomycin),- Cc k/sinh peptid & aa.Vd: Cc dn xut aa (cycloserin), cc cromopeptid(actinomycin),- Cc k/sinh d vng cha nit.Vd: polyoxin

- Cc K/sinh d vng cha O2.Vd: k/sinh polyete (monenzin).- Cc k/sinh mch vng no.Vd: Cc dn cht alcan (cyclohexinmid), k/sinh steroid (axitfuzidic).- Cc k/sinh mch thng:Vd: cc cht cha Phospho (phosphomycin).- Cc k/sinh cha nhn thm.Vd: cc dn cht benzene (chloramphenicol), cc cht nhnthm ngng t (griseofulvin), cc ete thm (novobiocin).

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Cu 2: Cc phng php p/lp vsv tng hp k/sinh?1)Ng/l chung:T cc ngun c cht khc nhau(t, nc, chtthi, bn,..) tin hnh nghin nh, pha long mu & cy vo cc

mtr nui cy chn lc ri cho ptr trn nhit thch hp, sau thun khit chng mi phn lp c, kt hp th k/nng sinhk/sinh ta thu c cc vss cn tm. Ty pp n/cu m ng dngling hot cc pp phn lp, th khc nhau nhm thu ly kt qumong mun.2) Cc pp:a) Pp cy dch chit t ln b mt thch: Cn chnh xc 1 lg

t (1-2g, t trng trt thg c ly su 10-15 cm) vo cooiss ri cho vo 1 lg nc v trng, nghin nh. Dng nc vtrng ri pha long thnh cc pha long thch hp(10-4 10-6). Nh dung dch pha long ln b/mt thch mtr dinh dngtrong hp pitri ri gt nh cho phn tn u. trong t m nhit thch hp (28 320C) cho ptr 5-7 ngy, xut hin cckhun lc ring r. Cy tch khun lc trn thch nghing thun khit ha, sau nui cy ri th hot tnh k/sinh of ccchng mi c phn lp.

b) Pp cy t trc tip ln b mt thch cha sn vsv kim nh:em reo cy trc tip cc ht t ln b mt thch tronghp Petri c mtr dinh dng cha sn vsv kim nh, t m24-48h ri mang ra quan st. Nu xung quanh khun lc ptr t

t xut hin vng v khun ta x/nh c trong ht t cvsv to ra k/sinh tiu dit c vsv kim nh. T khun lc tin hnh phn lp thu c vsv sinh tng hp k/sinh.c) Pp lm giu t: t trc khi c phn lp c cho vo ci thytinh, gi m xc nh, thnh thong cho t vsv kim nh vori trn u. trong t m 1 t/gian ri em ra phn lp theo 1

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trong cc pp nu trn. Bng cch ny ta thu c vsv i khngvsv kim nh (d lc u vsv i khng c rt t trong td) Pp b sung k/sinh vo mtr: phn lp x khun ta c th

cho thm k/sinh chng nm & VK vo mtr phn lp vi nng t 5-25g/ml. Cc k/sinh thg dng nh: tetracylin,neomycin, penicillin,. Nng k/sinh thm vo cn phai cduy tr thch hp, nu k chnh s ptr of x khun mc tiu cng

b c ch.e) P/lp vsv sinh k/sinh chng ung th: Cc k/sinh chng ungth va c t/dng tiu dit t/bo ung th, va c t/dng tiu dit

t/bo.. Nn sng lc cc vsv to k/sinh chng ung th ta cth dng vsv lm test th, cng c th s/dng cc chng nmmen b.

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Cu 4: Trnh by c ch tc dng ca khng sinh?- Cc k/sinh t/dng c bn wa vic c ch cc pn ng tng hprt khc nhau of t/bo vsv gy bnh. Chng lk vo cc v/tr

chnh xc hay cc p/t ch of t/bo vsv m to ra cc pn ngtrao i cht. Cc ch t/dng /trng cho tng nhm k/sinh,tuy nhin trong nhiu trng hp ng/ta vn cha bit chnh xcht.- C 6 mc t/dng khc nhau /vi t/bo VK or nm:+ T/dng ln thnh t./bo: tng hp murein (vancomycin,

phosphomycin)

+ T/dng ln mng nguyn sinh cht: thay i c/trc(polymixin, amphotericin) lm mt chc nng of mng lm chocc p/t c k/lng ln & cc ion b thot ra ngoi.+ T/dng ln w/trnh tng hp AND: phn chia (antracyclin),dch m(anzamycin).+ T/dng ln w/trnh tng hp Pr: ribosom(aminoglycosid gnvi receptor trn tiu phn 30S of ribosom lm cho w/trnhd/m ko chnh xc);+ T/dng ln s trao i cht h hp: antimycin.+ T/dng ln s trao i cht trung gian: sulfamid.

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Cu 5: Trnh by tnh khng thuc of vsv & nguyn tcs/dng khng sinh?

1) Tnh khng thuc of vsv: hiu v tnh khng thuc of

vsv trc tin ta phi hiu t/no l khng thuc. Khng thucl h/tng vsv mt di tnh nhy cm ban u of n trong 1t/gian nht nh hay vnh vin vi t/dng of k/sinh hay hatr liu. C 2 kiu khng thuc: Khng thuc t nhin &khng thuc mi nhn.

+khng thuc t nhin: l c trng ca tng ni vsv nht nhi vi 1 ks nht nh no . Tnh cht ny c sn trc khi sd

cc ks . Lin quan ti ph tc dng ca ks,c th c trngcho tng loi,cho tng nhm,tng sp. L thng tin di truyn csn trong NST. V mt sinh hc c 2 c ch tc dng: tnhthm ca tb v s thiu vng phan t ch.+khng thuc mi nhn: xut hin trong chn lc t nhin ccchng khng ca qun th vsv nhy cm khi sd ks.a) C ch d/truyn hc: Xt trn phng din khng thuc minhn c th l do thay i gen NST(B NST), c th do tipnhn gen plasmid(d khng plasmid hay ngoi NST)* Khng thuc do B NST: Kiu khng thuc ny chimkhong 10% tng s vsv khng thuc, x/hin ch yu /vi cck/sinh nh: -lactam, aminosid,B kiu ny t n nh & td/truyn.

* Khng thuc plasmid: kiu khang thuc ny rt p/bin, chimkhong 90% s vsv k/thuc. K/thuc ngoi NST l h/tng akhng thuc do nhn t R, nhn t R(gm 2 phn:phn x/dnhtnh k/thuc & phn truyn ti gen k/thuc) chnh l cc gen/khin tnh k/thuc. W/trnh truyn cht liu di truyn nhn tR theo 3 c ch:

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- G phi hp thit lp cu tip hp.- G bm v/liu d/truyn wa tip hp.- G ti t hp v/liu d/truyn t/bo nhn to thnh h v tnh

k/thuc of vsv.b) C ch sinh ha cc k/thuc mi nhn: c 4 kiu chnh:* Thay i tnh thm thnh t/bo:k/sinh ko thm wa thnh t/boc thi k pht huy t/dng(khng -lactam).* V hiu ha cc k/sinh=enzyme.* Thay i phn t chVd: thay i Pr ribosom lk vi streptomycin hay thay i ARN-

polymeaza lk vi rifamicin.* Hot ha con ng trao i cht mi khc m hot cht kt/dng.2) Nguyn tc s/dng khng sinh: Vic s/dng k/sinh cha hpl trong iu tr lm xut hin ngy cng nhiu vsv khngthuc. khc phc h/tng cn thc hin 1 s ng/tc sau:- Phn lp vsv gy bnh & th nhy cm of chng vik/sinh.Vd:pp khoanh giy lc,..- Ch s/dng k/sinh khi c nhim khun.- Phi hp k/sinh vi ch phm khc lm tng t/dng, gim

phn ng ph, bnh nhn ko t dng thuc.- Chn la ng k/sinh.

- S/dng k/sinh ng liu lng.- Nm vng chng ch nh of k/sinh.- Theo di hiu qu tr liu of k/sinh.

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Cu 6: Nu ng/l chung cc pp nh lng k/sinh, trnh bypp vsv nh lng khng sinh?1) Ng/l chung cc pp nh lng k/sinh:

- K/sinh c t/dng c ch s ptr of vsv kim nh, so snh vit/dng of k/sinh chun bit s tnh c h/lng k/sinh of muth- Da trn phn ng to mu of k/sinh or sp phn hy of chngri o bng my quang ph t ngoi or bng my so mu ritnh ra kt qu.- K/sinh khuych tn vo mtr thch cy vsv kim nh to ra

cc vng v khun. o ng knh vng v khun k/sinh chun& k/sinh th ri tnh ra hay tra bng thu kt qu cn phn tch.2) Pp vsv nh lng khng sinh:K/sinh c t/dng c ch s

ptr of vsv kim nh, so snh vi t/dng of k/sinh chun bits tnh c h/lng k/sinh of mu th. chnh xc of pp ny l5-10% .a) Pp pha long l/tc:- Dng nh lg k/sinh trong dch chit hay dch l/men. Phalong k/sinh trong mtr canh thang trong sut trong cc ngnghim v trng thnh cc nng khc nhau. C th phalong theo: 1:2; 1:4; 1:8; (dy hnh hc), hay 10:10; 9:10;8:10; (dy s hc),..- Ti mi nng k/sinh cho vo 1 lg vsv kim nh, sau

t vo t m 370

C trong 18-24h ri em ra c kt qu, nung no trong sut ti nng pha long k/sinh tiu dit vsvkim nh.- t k/qu chnh xc cn ch : thnh phn mtr pha long

phi n nh, lg VK kim nh cho vo cc ng nghim phibng nhau & t/gian cho vsv kim nh phi n nh.

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b) Pp khuych tn trn thch:* Nguyn l: K/sinh khuych tn vo mtr thch cy vsvkim nh to ra cc vng v khun. o ng knh vng v

khun k/sinh chun & k/sinh th ri tnh ra hay tra bng thu ktqu cn phn tch.* Pp khuych tn trn thch thg c s/dng nh lg k/sinh, cnch ti dy lp thch, pH mtr & t0C nui cy. Nhngk/sinh polypeptide kh khuych tn trn thch nn thg cho vomtr nh lg cc cht lm tng khuych tn of k/sinh(CaCl2lm tng khuych tn of k/sinh gramicindin C), or t hp

Petri c VK kim nh vo t lnh (40C) trong 2-6h k/sinhkhuych tn vo mtr thch ri mi em t vo trong t m chovsv ptr. Trong pp ny ta p/bit 3 bin thin khc nhau: pp trthp, pp c l thch & pp khoanh giy lc.

PP. ng tr thp PP. khoanh giy lc

Cu 7: Trnh by cc k/nng ng dng ngoi y hc.

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Cc nh n/cu & k/t kt hp vi cht ch vi nhau nhm pht huyti a tnh hiu wa of k/sinh.a) Khng sinh trong chn nui: K/sinh c dung nh cht k/thchtnagw trng gia sc, gia cm, gim chi ph thc n, k/thch tngsn lng trn g vt. Cc k/sinh:enduracidin,mikamicin,virginiamicin(peptid), tilozin, - K/sinh t/dng ln h sv ng rut lm tng s/lng vsv c chtrong rut, tng cng tng hp VTM, tng cng ti hp th thcn,k/sinh lm gim cc vsv c hi tit ra cht c or s/dngVTM, lm gim pH rut, Cc t/dng trn gip v tng cngtrao i cht nhanh.- Mt cch gin tip tng cng iu tit hormone /bit lhormone s/trng gip c th ln nhanh.

b) K/sinh trong trng trt: Vic chn k/sinh dit vsv gy bnhcy trng ko ch ch n t/dng k/sinh m cn phi w/tm nh/qu k/t.* K/sinh dng u tranh vi bnh thc vt phi tha mn cc yocu sau:- Hot tnh k/sinh mnh /vi mm bnh.- D thm vocc t/bo cy.- Liu iu tr k hi n cy.- K/sinh phi bn vngtrong 1 t/gian d b mt hay thm su vo trong cy.* u im of k/sinh trong b/v TV:- Liu rt thp nhng vn ct/dng.- D b cc vsv t phn hy.- Ko c or rt t c /vi Vmu nng & cc cn trng c ch.c) K/sinh trong cng nghip thc phm: Bo qun thc phm ti& cc thc phm ng hp l vn rt w/trng. Cc k/sinh cs/dng trong bo qun thc phm ph thuc vo cc quy nh lut

php of nh nc.vd: 1 s cht bo qun nh: Nizin cht l tng b/v phormat,c chua, u xanh, bp ciCu 8: Trnh by cc ng/liu cht dinh dng trong sx k/sinh?

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Mi trng dinh dng nui vsv phi cha all cc ng/t cn thitcho t/bo s/trng & sinh tng hp di dng d hp th & tip nhn.Cc ng/liu phi tha mn yu cu l d kim, chuyn tr thun tin& c sch cn thit.

a) Cc ngun cacbon: Cc loi cacbonhydrat l ng/liu truyn thg ofcg nghip k/sinh & VTM.

S/dng cc loi ng sch lm mtr: sacaroza(lmen sx Fumagillin),glucoza, lactoza(sx penicillin),

T/bt & Dextrin l cc ng/liu lmen k/sinh tt /vi cc chng vsv ck/nng to ra cc enzyme amylaza. C th s/dng t/bt vi cc ngungc khc nhau: ng. Khoai\, sn,

Cc loi du thc vt(du u tng, du c,)c th dng nhngun cacbon h tr.b) Cc ngun Nit: C th s/dng cc mui amoni [(NH4)2SO4,NH4NO3], mui nitrat[NaNO3, KNO3], lm ngun nito ty thuc vochng vsv. Cao ng: l ngun nito d hp th (cn thg s/dng 50% chtkh) l w/trnh ph trong sx t/bt ng.Cao n/men: l ngun m c nhiu loi vsv a thch. Cao n/men c sxt nm men bnh m l chnh bng pp t phn (ahttolysis) 50-550C,hay bng plasmolysis vi nng cao or bng thy phn acid nh(pH=5-6 vi HCl). Cao n/men cha cc acid amin, cc peptid, ccVTM tan trong nc & cc loi cacbonhydrat.Bt u tng: c s/.dng nhiu trong l/men k/sinh do hp th tg ichm & ko xy ra cc c ch /khin trao i cht.c) Cc cht khong, vi lng & k/thch:

/bo cho vsv ptr b/thng cn phi /bo cc cht khong nh:

NaCl, K2HPO4,Bn cnh cc cht khong phi b/sung 1 lng nhcc ion vi lng cho vsv. Tuy nhin nng cn thp, nhng k v thm xem nh v/tr of chng.

i khi cn b/sung them vo mtr dinh dng 1 s cht k/thch ptrnh: VTM,

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Cu 9: Cc pp ci to ging(CLTN, B nhn to) & trnhby cc k/nng trong ci to ging?1)Cc pp ci to ging:

a)b t nhin: cc vsv b b theo tun xut khc nhau, nncoong vic ci to ging y l cn phi tuyn chn ly ccc th c hot tnh cao nht n/cu tip theo. y l pp chn cc chng c h/tnh cao, tuy nhin bng pp sng lc cc cth b t nhin c h/tnh cao ch n/cu ban u, k c gi trp dng vo sx.

b) b nhn to: Cc chng vsv chu t/dng of tc nhn gy b

mnh nh tia X, UV hay cc tc nhn ha hc nh: etylenimin,dimetylsulfat,..phn ln cc vsv b git cht, cc c th cnsng xut hin nhiu dng b or gim (mt) kh nng sinhk/sinh (b m tnh) or lm tng k/nng sinh tng hp k/sinh(bdng tnh).2) Cc k/nng trong ci to ging:- T chng vsv sinh ra 1 loi cht c c/trc tg t ng/ta to b &

phn lp cc b ch to ra 1sp cn thit lm tng cao sn lng& lm cho qu trnh tinh ch k/t hn.- Lm gia tng sn lng hot cht m/tiu nh ng dng 1 cchlinh hot cc pp b & sng lc hay ti t hp, sang lc da vokinh nghim hay h/ng c thc bng c ch cc con ngsinh tng hp nht nh hay bng vic thay i h/thng

/khin.- a vo s/dng cc chng b c tip or tin phn ng thayi to ra cc k/sinh ng ng, c c/trc thay i so vik/sinh bit. Adramycin l cht c t/dng chng ung thmnh hn, l k/sinh antitumor c ng dng rng ri hin nay.

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Cu 10: Cc tc nhn b & cc kiu b trong ci to gingvsv sn xut k/sinh?a)Cc tc nhn gy b:

* Tc nhn vt l: Tia Rnghen(tia X), tia cc tm bc song 250-280 nm c t/dng gy cht mnh, t/dng gy b khi x/l ccchng vsv. Ngoi ra cn c th s/dng tia nowtron,, ngynay tia cc tm bc song di (=380 nm) kt hp vi ha chtcng thng c s/dng.* Tc nhn ha hc:+ N-mustar: tris-(-clor-etyl)-amin or Metyl-bis-( -clor-etyl)-

aminCH2 CH2 - Cl

CH3 N to c k/qu mong mun all cc nhn t ny u phic ti u ha. Phng php hu hiu nht l lp ng congliu t/dung kt hp vi xy dng biu tn sut.

Tuy nhin cch ny i hi rt nhiu cng sc, nhng li l conng duy nht em li kt qu.2) Tuyn chn b: (ch n/cu chn lc ngu nhin)* S tng trng to ln of cc vsv trong cng nghip k/sinh as u t c bng cch sng lc ny, tc l tuyn chn sau b.* Ng/l chung: Sauk hi x/l vi t/nhn b & nui cy ptr, trongs cc c th (clon) sng st c la chn ngu nhin ta tinhnh kho st k/nng to h/cht of chng.* Pp l/men m hnh c ng dng khi m cc tham s l/men (mtrdinh dng, cc k ngoi cnh) c duy tr ging k cngnghip. Cc chng tt nht thu c bc ny li em b & snglc tip. Sau 1 lot cc bc b & sng lc phn lp sx s tngln dn dn. Pp ny cn gi l tuyn chn bc thang hay b bc

thang.* Trong ci to ging cng nghip khi kho st cht lg toh/cht of cc c th (clon) sau cc bc b mnh xut hin 3 tnhth sau:- Cc clon c sn lg cao hn of b m mc va phi xuthin vi tn sut cao hn & d xc nh hn.

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- Cc clon c sn lg cao hn of b m nhiu xut hin vi tns thp, do vy tm kim chng rt mt thi gian.- Do phi s/dng tc nhn b liu lg mnh thg xut hin b

nhiu tng lm hiu sut k tng tip c na. trnh mt thi gian tm kim chng c hiu sut cao nht taly 5-10 chng c mc tng trng va phi lm chng bm & tip tc b tip(trnh c b nhiu tng).* Tn s xut hin cc tng trng sx ph thuc vo chng vsv,k b cng nh vo q/trnh sinh tng hp & /khin of h/chtcn ti u. Thng thng theo chu k b phi kho st t vi

trm n vi ngn c th.Cng sut sng lc s quyt nh vntc tin ln pha trc of ci to ging. tng cng sut sang lcng/ta xy dng cc h/thng t ng kiu Glaser Dumb-Waiter hon ton t ng.

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Cu 13: Trnh by cc pp gi ging vsv?- Gi ging nhm gi gn k/nng sng st & cc tnh cht cgi tr k/t k thut of cc chng vsv sx l ht sc w/trng

nhng li tn nhiu cng sc & kh khn.- Gi ging trng thi tim sinh gip duy tr c cc t/cht banu of vsv vi s chm sc ti u.- La chn pp gi ging trong mi trng hp ph thuc vok vt cht, k thut of phng th nghim, sc sng of vsv, tnh

bn vng dt hc & yo cu s/dng sau nay.a) Gi ging trn mtr thch: L pp c ng dng kh rng ri

trong thc t, mc d k phi l k/t & ng tin cy nht. Sau 1chu k t/gian nht nh phi cy truyn chng vsv sang mtrmi, cho ptr ri li ct gi. Vic ny i hi tn cng sc &a n nguy c thoi ha chng dn n gim thiu of mt inhng t/cht c bn of chng.

Trong gi ging /bit trn mtr giu cht hu c c th xuthin phagolysts t pht chng vsv. Nhim phage c th lmgim or mt h/tnh of chng. lm gim kh b/mt thch cth trng paraffin nt or bc mng nilon hay mng polyetylen,mt khc c th dung mtr 1% du thc vt.

Gi ging trn thch c th duy tr nhit -20 -220C .Trong thc t ng ta gi bo t or t/bo s/dng 48h tui trongmtr glycerin 15%. Phng php ny n gin, nhng li nm

trong k/nng cht of vsv t cc t/b ng lnh.b) Gi ging trn ct v trng: Ly ct song ra trong dngnc chy ti khi ht bn, sy kh ri un nng trn ry st,sau ry wa sang nh ri cho vo ng nghim hay ampul,ng nt li ri kh trng 1200C trong 1h trong ni hp.

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T 1 lg ging c th s/dng lm nhiu ln trong t/gian tg idi.c) Gi ging trn t v trng: Ly 2g t vn, nghin nh,

cho vo ng nghim ri them 2-3 git nc, lm nt & khtrng 1200C trong 1h, sau m 300C trong 24h ri khtrng nh trn 1 ln na. Sau khi th v trng, cc ng tc cy vsv & c bo qun. T 1 lg ging c th s/dng lmnhiu ln trong t/gian tg i di.d) Gi ging trn ht ng cc: T ht k c vng cn ly 1kgcho vo bnh, them vo 800ml nc si, un si trong t/gian

nht nh cho k no nc, sau y lm nt & ng 30 riem ra mt bn lau cn etylic cho ngui. Cn ly ra 15gvo bnh nn 250ml ri kh trng 1150C/40 . Th v trng,

bnh k v trng c ly cy 2ml hn dch bo t trong Tweenhay 3ml si dinh dng. Ni cy trong 2 ngy m trn mylc, ty tng chng m k c cho ptr 1 tun 280C or phi lc ph v khi k hay long si. Vsv ptr n/vy c em sy chnk 5-10 mmHg trong 3 ngy nhit 250C cho n khi khi lgcc bnh k k thay i na, m ht k k thay i 8% . Cc

bnh cn li c trng paraffin nt & ct gi nhit phng &ni kho ro. C th s/dng nhiu ln sx k/sinh.e) Gi ging bng ng kh: ng kh c s/dng rng ri gi ging vsv t/gian di. Pp ng kh da trn thng hoa nc

di chn k t cht liu ng lnh. Vi m/dch ny phichun b hn dch bo t m c trong huyt thanh nga k ccht bo qun or trong dung dch cha 10% sacaroza & 1%gelatin, pH=6,8 7,0. Phn 0,2 ml hn hp dch bo t vo ming ampul(l thy tinh trung tnh c ch tht) bng pipet.Ampul c lm lnh xung bng hn hp CO2 bng & etanol ( -

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70 -800C) trong 5 & xp vo my chn k lm kh psut 100mmHg. T/gian lm kh ph thuc vo chn k, b/mt

bc hi & th tch cn lm kh. m cn li k vt wa 3-5%.

Ly ampul ra khi my chn k, c ct gi nhit 50

C thunli hn.S/dng pp ny s: Chng c bi nhim,duy tr c cc /tnh tt

of chng & tit kim c v/tr.f) Duy tr h/tnh k/sinh of chng sx: duy tr h/tnh of k/sinh

bban u of chng sx th k th c gii php ton din ch bngcc pp bo qun, tn tr ging, m phi p dng cc pp phc

hp bao gm: la chn pp tt nht, nui cy chng sx trn mtrgi ging thch hp nht kt hp vi vic a vo ng dng cc

pp tuyn chn l/tc cc dng c h/tnh cao nht.

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Cu 14: Qu trnh l/men gin on, cc pha c th & cctham s /trng (tg, x, ) ?1) Qu trnh l/men gin on: Hay cn gi l ln men m,

l/men chu k & h/ng nh 1 h/thng. Ti thi im t=0h, tacy ging vsv to hot cht vo mtr dinh dng trong bnhl/men 1 cch v trng, ri tin hnh l/men trong /kin kh nnti u. Ngoi cp O2, cht ph bt, cht iu chnh pH th trongthi gian l/men ta k t/ng, bsung g thm vo h/thng. V vyq/trnh l/men thnh phn of h sinh khi vsv, nng cc sptrao i cht bin i k ngng. theo st cc din bin of qu

trnh l/men ng/ta tin hnh phn tch cc mu dch l/men mily x/nh tham s chnh.2) Cc pha c th & cc tham s c trng : 4 pha c thtrong l/men gin on.* Pha lag: y l pha ptr tim tang vsv thch nghi vi mtrmi. Trong vic hnh thnh pha lag ny trng thi sinh l ofging c ngha quyt nh. Cm ng to h/cht trong vsv k

ph thuc vo bn cht ngn hay di of pha lag.* Pha log: Cui pha lag l pha log, pha ny ptr nng ng nht= 1/x * dx/dt* Pha dng hay pha n nh: Sau khi ng ha cc cht dinhdng hay sau khi tch ly cc sp trao i cht, s/trng of vsvgim xung hay h/ton ngng li.Do s t phn of 1 s t/bo, 1

s cht dinh dng mi (cacbonhydrat, Pr) c gii phng, iuny a li s ptr chm of 1 s t/bo sng st. C rt nhiu sptrao i cht c gi tr c to thnh.* Pha suy tn: Cc cht dinh dng b cn kit, nng lg of t/bogim n ti thiu. t/bo cht i nh trong s/trng trong thik s/trng.

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/trng of pha ny l ph thuc tng loi vsv & vo cc ppnui cy c ng dng.

Trong 1 s trng hp vic sinh tng hp h/cht tuy cha

dng hn, nhng chm li & vic duy tr tip tc qu trnhl/men k cn k/t na v th cng cn kt thc l/men.

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Cu 15: Qu trnh l/men bsung: ng/l, cc cng thc chnh &ng dng?

Q/trnh l/men y cng bt u l/men 1 cch gin on, sauo ti 1 thi im nht nh bt u bsung. T/gian l/men do th tcht/b & tc bsung xc nh. Thg khi bt u ch s/dng 0,5 0,6V tng th tch, sau bng k thut fed batch tng ln 0,7 0,85 V th kt thc.

Pp c bsung khi bt u l/men cc t/phn nguy hi of mtr c ngdng vi nng thp hn so vi pp l/men m rt nhiu(thp hnnng cho tng loi vsv). Cc cht ny trong q/trnh l/men c

bsung l/tc vo mtr l/men thnh tng phn nh. Nu trong q/trnhl/men k th o c 1 cch trc tip & l/tc hm lg cht dinh dg th cn phi tm cc tham s gin tip l/wan & wa o of n tnhra h/lng cn bsung.

Pp c ng dng trong sx penicillin, acid glutamic, ng dngtrong cc trng hp: nng c cht kh thp nhng li cn nnh(l/men nm,), nng c cht kh cao & k thay i (l/menacid xitric), bsung tin cht lin tc (penicillin, indol,)

Cc cng thc chnh:

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Cu 16: L/men lin tc chemostat: ng/l h/ng, hnh nh qua th, cc cng thc /trng?* Ng/l: khi iu khin Chemostat trng thi cn bng (dng,

steady state) ng sinh trng cc t bo vi sinh vt c duytr bng thay i nng ca cht dinh dng ang c ch no. lm c vic ny bt k cht dinh dng no (cacbon,ngun Nito, mui, O2,...)* Hnh nh:

* ng dng:- Sn lg cao hn.- Nhiu k/nng o & iu khin.- Trong trng thi k i vsv ptr cn i( sinh khi ng nht).- Chemostat ng dng ch yu trong l/men SCP, nm men bnhm.- Chemosta ng dng trong sx etanol, bia.

22

Sjx

Dmj

X

D

S

S0

D

S.xX

2

X1

S03

S02

X3

S0

Bin thin cc tham s c trng ln Chemostat theo D


23/38

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Cu 17: L/men lin tc turbidostat, l/men bn l/tc & ngdng?a) L/men l/tc turbidostat:

Khi l/men dch l/men c 1 my bm chu k ngoi vng wa c quang hc (OD = Opitical Densty) of dch l/men o c l/tc.Qu trnh l/men c bt u gin on.

Do phi o l/tc quang hc m pp ny ch c th s/dng vsv 1t/bo & cc dch l/men tg i long.K thut turbidostat ch yu n/cu: ti u ha, ko di tng trngly tha.

23

X max

X min

X

t

t

Hnh nh ln men turbidostat

t1

S2 t2 S3

t3

Nguyn cu ti u ha vi turbidostat

t

X


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b) L /men bn l/tc & ng dng :y l bin tng l/men khi vic rt bt dch l/men & bsung

them mtr dinh dng k xy ra l/tc m ch nh k trong 1 s thiim nht nh.

L qu trnh c li v tng c lg dch l/men cha h/cht mc tiu(nu vic bsug cht dinh dg k lm gim hiu sut to sp). Tuynhin thi im bsung & ly bt dch phi da trn tnh ton cngnghip hp l.

24

tX

min

Xmax

X

t

Hnh nh ln men bn lin tc


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Cu 19: Trnh by cc h/thng bnh l/men: ng/l h/ng, ngdng my tnh in t trong l/men sx k/sinh?a) Cp kh vi cc b phn vn ng: y l loi bnh l/mensc(cp) ko kh kt hp vi khuy trn, c ng dng rng ri nht& cng ptr mnh m nht.

Trong thc t bnh l/men cp kh c khuy trn l thng dng

nht.b) Cp kh vi my bm vng ngoi:

L loi l/men ct bt kh, hi lu tia hay tia chm cng kh thngdng.

25

C khuy khuy ng dn khuy t ht


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T/dng cp kh of p sut kh c t/dng chuyn ng dng chtlng nh my bm tng cng thm nn h/qu t c cng khcao.c) Cp kh vi p sut kh & pha kh ng:

L loi hnh l/men kh thng dng trong cng nghip, nhng ng dng loi hnh ny trong nhiu trng hp phi ci to c cc

bin chng ging cho nht dch l/men thp. Nh vy m gimc chi ph nng lg, nng cao c tnh cnh tranh cao of cng ngh.

Cp kh vi pha kh ng l h/thng /trng cho l/men b/mt khidung kh nn hay qut gi cp kh cho bung l/men.

Re = NRe = Di2 . N . P Di: g knh my sy (cnh khuy) (cm).

N: vn tc my sy ( v/s, s-1).P: khi lg ring (g/cm3).: nht ng (g/cm.s).d) m bo v trng:

Phi /bo 1 s quan im v kt cu: bnh l/men ch c t cam, l nhng ca ti cn thit, c b dung vng m, ca lndng dong lt.

26

Ct bt kh airlift


27/38

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Nn trc l h/thng dong trt (Simmering), kh trng bng hi& sau dng nc ngng ngng hay dng glycerin gi.

Qu trnh l/men v mt ng/tc kha hi phi loi h/ng l/tcnhy m.

Cu 21: Cng sut my khuy of bnh l/men c cp kh?* Do sc ko kh vo bnh l/men m hnh thnh h kh-lng

lm gim khi lg ring, do o lm gim yu cu cng sut.Cng thc(3.62) [Michel & Miller](t vit)

27

Hi ra

Hi vo

Kha hi v hot ng


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Thng s: Pg cng sut my khuy c cp kh.K - Hng s (kt cu) (0,706 / V).P1 cng sut my khuy k sc ko kh.

N vn tc my khuy.D1 ng knh my khuy (cm, dm.m).Q - vn tc cp kh (VVM, L / L.min, m3/m3.min).

* Trong thc t cng nghip khi th tch bnh l/men tng, yucu v cng sut my khuy gim.

Th tch (m3) Cng sut(Kwh/m3)

0.1 11 - 131 5 4 7

15 - 20 1 3*Nu ko c my khuy ta co cng thc: t vitThng s: F dng cp kh ().

P p sut ko kh .Vo tc kh ti b chia.Pc - p sut kh ti b chia. 0.06

Cu 24: Trnh by qu trnh kh l/tc mtr l/men.a) Kh trng l/tc dch l/men:* Mtr dinh dg sau khi s un nng c a vo b phn un nngtc thi vi hi c bm trc tip nng nhit ln 130-1400C, dch

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dinh dg c chy trong rut xon b phn duy tr t 2 3, qua vangin n x tc thi sang b phn lm lnh h nhit xung 800C& c lm lnh xung nhit l/men nh mtr lnh trong b trao inhit thng.

B kh trng lin tc trao i nhit tm* U im of pp:- D /khin & t ng ha.- Trn nhit kh trng cao hn, thi gian kh trng ngn hn,

b/m an ton hn & du nh.

29

Lmlnhs b

Lmlnh

ungnng

Nc lnh unnngs b

Mitrng

hi

Mt vtrng

t(min)

140020 S

2-3 pht

20 S


30/38

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- Qu trnh l/tc cho dinh dg ng nht hn ( tng cng hiu sutl/men).Pp kh trng ang ngy cng c ng dng rng ri trong cngnghip.

b) Kh trng ko kh:

- Cc ht bi l lng trong ko kh & s vsv giao ng rt mnh phthuc vo v/tr, chuyn ng cng nh vic x/l trc. Ngoi tric khong 10.000 ht/m3 , trong khong 2.000 vsv/m3 , 50% svsv l bo t nm, 40% l VK.- Cc bnh l/men c vn hnh vi vn tc cp kh 0.5 1.0 VVM(m3/m3.min). Nu th tch bnh l/men l 50m3 & vn tc cp kh l1.0 VVM th phi cp 3.00m3/h k kh v trng.

- Trong cc pp hnay c cng nghip ng dng pp lc v trng.Cu 26:trnh by cu trc ho hc ca cc ks penixillin tnnhin,sinh tng hp,bn tng hp.Khung c bn ca pen l 6-APA (6- amino pen axit) c nhnthiazolidin v vng -lactam

30

Chn ko

Mt v trng

Phn mt

M trng

hi

Kh trng mt lin tc-h bm hi


31/38

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Trong sx nu k thm cc tin cht vo trong dch ln men thicc ks t nhin c hnh thnh.Vit 1 cch tng qut R=OOC-CH(NH3)-(CH2)3-CO-

V nu thm cc tin cht vo dch ln men thi c cc ks tnghp. Sd cc phn ng ho hc gn 6- APA vs dx R th c ksbn tng hp.tn Gc R1Cc khng sinh t nhin2-pentenyl-pennixilin(penF)

n-amyl-pen(dihydro-penF)n-heptyl-pen(pen K)

p-hdroxy-benzyl-pen(penX)

pen N(D)iso-pen N(L)

benzyl-pen(pen G)cc khang sinh tng hp

phenoxy-metyl-pen(penV)

allyl=mercapto-metyl-pencc khang sinh bn tnghp

propicillin(bn ax,khng-lactam)

CH3-CH2-CH=CH-CH2-CO-

CH3-(CH2)4-CO-CH3-(CH2)6-CO-

-OOC-CH(NH3+)-(CH2)3-CO-

CH2=CH-CH-S-CH3-CO-

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ampicillin,dit G-,bnax,nhy cm penicillinaza

cu 38: trnh by sx vtm B12 nh Ps.denitrificans v qutrnh tch chit tinh ch vtm B12a-ln men sx vtm B12sx vtm B12 chng nh Ps. L pp rt kinh t.S sx vtm B12 nh chng Ps.

32


33/38

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Ging ct gi=>mt A(hot ho ging 4 ngy,28C)=>(bnh nn2l;0,6l mi trng;my lc)ging cp 1, 28C,48h(mt E)=>(25-50l/40-48l,vkh=200rpm)ging cp 2,32C,25-30h;0,5VVM(mt

E)=>(300l mt/500l tng,cp kh:0,5 VVM,khuy:150-200rpm)ln men,32C,140-160h(mt P)Trong qu trnh ln men cn quan tm ti ho tan caoxy,trong g u(20-30h) cn duy tr ho tan cao,n sang gsau li cn oxy thp(xp x 0) s c li cho sx.

b-qu trnh tch chit tinh ch vtm B12c nhiu bin thin trong qu trnh tch chit v tinh ch vtm

B12dch ln men Ps. + un nng 30 120C,lm lnh chnh pH v 8.5

+thm KCN,khuy16h,25C

+thm ZnCl2 200g+chnh pH 8+thm cht tr lc

200g,khuy lc

Dch lc chit 3 ln vi

350ml hn hp cresol:CCl4(t l 1/2)

Dch chit hu c 1 thm 500ml n-butanol

Chit 3 ln vi

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100ml nc

Dch chit nc chit 3 ln vi hn

hp cresol: CCl4 t l 1/2

Dch chit hu c 2 thm 200ml axetonv 100ml ete, vtm B12 th ta xung

VTM B12 th ho trong 10mlmetanol

Sc k trn oxitnhm hot ho 4gPhn hp ph vs

a.axetic 2% trong metanol,ta ete

VTM B 12 kttinh

Cu 37:cng thc cu to v tc dng ca VTM B12,ln men sxvtm B12 nh P.shermanniiCng thc cu to:-cha khung tetrapyrrol.trong cobalanmin ti C1 ca ribose lk vs5,6-dimetyl-benzinmidazol. Bn cnh CN- cha CO + trung

34


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tm. Thay CN- = cc nhm khc nhau,ta c cc dx khc nhau. Ccng ng vtm B12 vs cc bazo d vng khc(cc

purin,benzimidazol th) c cc vsv to ra t pht hay khi cho thmcc cht y vo mt.Tc dng ca vtm B12C vai tr quan trng trong ti to tb,tb thn kinh, phc hi cc nhum gan,tham gia to hng cu,vtm B12 tham gia tng hp DNA v1 s cht khc: cholin metionin,c trong gan ,sa trng.Thiu vtm B12 c th b suy nhc ton bVtm B12 hu nh k c c tnh.Trong iu tr c th dng vtm B12 :-iu trn mu c tnh: c th dng phi hp vs cc thuc khc(axfolic, ax ascorbic)-iu tr cc bnh v vim thn kinh.

Cu 32: ln men sx ,tinh ch ,tch chit streptomycin-ln men sx streptomycin:Trong LM sx streptomycin ging S,griseus c nui cy 26-28C/48h. Ging cp 2,3 c nui trong 24h pH 6,5-7

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i vs chng S.gieus DTH -2 mt nhn ging v LM c cc thnh phnsau:Mt nhn ging:Glucoza 40g, dextrin 2g,bt u tng 20g,(NH4)2 SO4 5,5g,CaCO3

6,5g, NaCl 2g,KH2PO4 50mg,pH t nhinMt ln men:Glucoza 50g,dextrin 10g,bt u tng 30g,(NH4)2SO4 9g,NaCl2g,CaCO3 9g,cao ng 4g, KH2PO4 25mg,du u tng 7mg/l pH=7,2Sau khi time ging chuyn Sng mt ln men v ln men 26-28C,Ph 6,8-7,cp kh 1 VVM,khuy lin tc 4-7 ngy m.

Tch chit:Sau khi ln men kt thc,tin hnh lc loi sinh khi. Thm axoxalic,loi ion Ca2+,chnh pH v trung tnh ri em lc loiCa(C2O4)2. S dng nha trao i ion cationit yu ang chu k Na+ hp ph streptomycin theo phn ng:R-COONa+ +(streptomycin) => R3(strep) + 3Na+Nha trao i ion c th l Amberlite IRC 50, Woffatit CP300....saukhi ct bo ho dng ax HCl 1N phn hp ph streptomycin ra khinha theo phn ng:R3(STREP) + 3 H+ => 3ROO-H + strepPhn on chnh c c chn k.dch c c c cho thm metanol laoij NaCl i dng ta ri m lc. Thm CaCl2 vo cha metanolv mui kp str.clohydratcanxiclorid kt tinh s c em lc. T muikp cho Streptomycin c cho tc dng vs H2SO4 to mui sunfat. Saukhi tu mu v c li n nhit cn thit th tin hnh lc v trng.

Dch lc v trng c em ng kh. Sp thu c t tiu chun dcim.

Cu 25: ng dng my tnh in t trong ln men sx khngsinh.

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Trong LM sx ks sd my tnh in t trong 2 linh vc:+ti u ho vs s tr gip ca my tnh:tnh ton cc m hnhkt hp x l cc kt qu,x td cu cc tham s ln qt LM

+kim sot,iu khin qt LM :trong cng ngh LM cc nctin tin kt ni my tnh iin t on-line LM tr thnh hinthc

CU 27 trnh by con ng sinh tng hp v iu khin sxpen trong P.chrysogennum

37


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Vng- lactam-thiazilidin ca pen c xd t L-cistein v L-valin.Con ng sinh tonhr hp bt u t ngoi ribosome t L--AAAto dipeptid vs L-xistein ri qua tripeptid vs L-valin m tip tc, sau1 s bc kt hp ng vng c th tch c isopen N. Pen G c tothnh khi L--AAA i sang nhm phenyl axetic. Cn 6-APA k

pai l sp trung gian m ch do thiu tin cht m cha c loi ra.

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