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LI M U
Lysine l mt trong chn loi amino acid m c th ngi v ng vt khng th t tng hp c, l mt trong 12 axit amin thit yu cn c trong ba n hng ngy. Lysine l thnh phn ca nhiu loi protein, l yu t quan trng trong vic duy tr h min dch, n gip tng cng hp th v duy tr canxi, pht trin men tiu ha. Tuy nhin, c th ch tng hp c Lysine qua con ng thc phm nh lng trng g, tht, c, cc loi u v sa ti hoc di dng thuc. Nhng vic cung cp Lysine qua con ng thc phm rt d b ph hy trong qu trnh un nu. v vy, cch cung cp Lysine hiu qu nht l s dng cc loi thc phm ch bin sn, c b sung Lysine vi liu lng thch hp, va cho nhu cu ca c th. Lysine c tng hp t cng ngh ha hc, tip l cng ngh tch chit v ph bin hin nay l cng ngh ln men vi sinh vt. v cng ngh sn xut lysine trn th gii, lysine c sn xut quy m cng nghip cc nc nh M, Nht Bn, ng u. Cng ngh ln men Lysine ngy cng m rng vi hiu sut qu trnh ln men tng i cao. Ti Vit Nam, hng nm lysine c nhp khu vo vi s lng tng i ln. Tuy nhin ch duy nht cng ty VeDan l a vo sn xut vi quy m cng nghip, ngoi ra cc nghin cu Vit Nam mi ch dng li quy m phng th nghim, ging vi sinh vt cha t chun. ti thc t ti phng nui cy vi sinh vt cng ty CPHH VeDan VietNam, nhm gii thiu v quy trnh cng ngh sn xut Lysine bng phng php ln men quy m phng th nghim. Th nghim trc s lm tin cho th nghim sau, t gip chng ta tm ra phng php v iu kin nui cy ti u p dng vo sn xut quy m cng nghip, nhm mang li gi tr kinh t cao nht.

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CHNG 1: TNG QUAN V CNG TY CPHH VEDAN VIETNAM

Lch s thnh lp Tn cng ty: Cng ty CPHH VEDAN VIETNAM Cng ty Vedan vietnam xy dng nh my vo nm 1991 ti x phc thi, huyn long thnh, tnh ng nai. Nm ng nam Thnh Ph H Ch Minh, tng din tch 120 hecta. n nay, cc hng mc c a vo sn xut gm: nh my xt-clo, nh my bt ngt, nh my tinh bt, nh my tinh bt bin i, nh my lysine, nh my pht in v hi, ba h thng s l nc thi hin i, h cha nc cho sn xut 60.000m3, cng Phc Thi vedan, ng giao thng chuyn dng, cc khu lm vic, sinh hot, vui chi. Hin nay, VeDan Viet Nam c th thay th cho VeDan Dai Loan v tr thnh nh my sn xut ln nht trn th gii. Chin lc kinh doanh v pht trin Vi mc tiu, kinh doanh lu di ti vit nam, cng sng to tng lai. Cng ty VEDAN VIETNAM lin tc u t, p dng cng ngh tin tin, tch cc thc hin cc k hoch nghin cu, pht trin. thng xuyn hp tc rng ri vi cc c quan, hip hi nghin cu v cc t chc o to bi dng ngun nhn lc c y tim nng hon thnh xut xc cng vic, ng thi to ra bc u tt p cho vic nghin cu pht trin ca vit nam. p ng nhu cu v tnh cnh tranh trn th trng, cng ty vedan viet nam lin tc tuyn dng cc chuyn gia trong v ngoi nc. Lin tc nghin cu v pht trin nhng sn phm mi, tt c nhng thnh qu tr tu ca s n lc khng ngng ca ton th cn b cng nhn vedan c n p bng s khng nh cht lng v uy tn dng sn phm vedan trn th trng trong v ngoi nc. Mc tiu kinh doanh ng tm hip lc, pht trin lu di

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Nhm ton cu, lm ti ch Ln mnh, ln mnh hn na.


1.1.

Nng lc sn xut Ngay t khi mi thnh lp, cng ty VEDAN VIT NAM khng ngng

i mi cng ngh v nng cao cht lng sn phm bng vic u t thay th cc thit b c, lc hu, nng sut thp bng thit b tin tin. Trong sut nhng nm qua, cng ty khng ngng tuyn dng v o to nn nhng nhn vin u t p ng vi nhng nhu cu m rng sn xut v kinh doanh ca mnh. Vi s lng hin ti khong 2700 nhn vin, trong hn 30% l trnh trung cp tr ln, Vedan ang v s to ra nhiu sn phm cht lng hn na c th p ng nhng nhu cu ca ngi tiu dng. ng thi Vedan lun n lc nghin cu v pht trin cc sn phm mi p ng nhng i hi v nhu cu ca th trng trong v ngoi nc. 1.5. Sn phm Vedan 1.5.1. bt ngt Sn phm bt ngt ca cng ty vedan khng nh c uy tn v cht lng sn phm i vi ngi tiu dung trn ton th gii trong sut nhng nm qua. Sn phm c chng nhn cht lng ISO 2002, vi thun khit t trn 99% t mi tiu chun ca nh nc v an ton v sinh thc phm. 1.5.2. Acid glutamate L loi acid amin c tng hp trong c th ngi, chng c s dng rng ri trong tr bnh v b sung sinh trng cho thc vt. ngoi ra acid glutamic c hai gc hydroxyl v mt gc amin, n l mt cht lng tnh mang c tnh acid v kim, c th lm nguyn liu cho cc loi thc phm, ha phm v m phm.
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1.5.3. Axt amin Trong hn 20 loi acid amin cu to nn cc t chc c th, trong c 8 loi c th khng th t tng hp c m phi tip nhn chng t thc phm. do yu cu c bit , Vedan cho ra i sn phm Lysine c p dng rng ri trong thc phm nh cht tng cng dinh dng, cht ph gia thc phm, dch truyn amin..v.v. 1.5.4. Tinh bt khoai m Tinh bt khoai m vedan c s dng rng di trong nghnh thc phm nh lm cht n, cht kt ni, cht n nh, cht lm c. ngoi ra nh my ch bin tinh bt bin tnh ca vedan cng nghin cu v pht trin cc sn phm t tinh bt c ng dng trong cc nghnh cng nghip khc nh: dt, sn xut giy hay xy dng ..v.v. 1.5.5. Thc n chn nui Qua thi gian di nghin cu, Vedan sn xut loi thc n ng vt mi, cht b sung thc n chn nui vedan mt r ln men c c, sn phm ph t dung dch ln men ca mt sau khi ln men, khng nhng c cc thnh phn dinh dng ca mt m cn c thm protein. Sn phm c s dng rng ri cc nc u, M. 1.5.6. Phn bn hu c Phn bn hu c Vedagro c ch bin t tinh bt m v r ng bng cng ngh vi sinh hin i. Sn phm c hi ng khoa hc b nng nghip v pht trin nng thn cng nhn l tin b k thut, cho php s dng trn cc loi cy trng cn. 1.5.7. Acid clo hydric/ xt

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Nh my xt-clo ca cng ty c quy trnh sn xut tin tin v ch qun l cht ch, do vy sn phm thun khit, t tiu chun thc phm. Acid clohydric, xt lng v Hypoclorite ca cng ty khng nhng c dung trong nh my sn xut bt ngt, cn l nguyn liu c bn cho cc nghnh sn xut. hin nay, cc nhu cu ca th trng pht trin mnh m, i hi vedan phi c gng khng ngng pht trin kinh t v x hi Vit Nam. 1.5.8. Hypochlorite NaClO. L mt cht oxi ha mnh, tn ti trong dung dch c c, khng c tnh kim. gp nh sng v nhit s nhanh chng phn hy.c ng dng trong ty trng vi, qun o, tinh bt, ti sinh giy, x l nc thi, x l v sinh..v.v.
1.6.

Tuyn dng v pht trin Nhn ti bao gi cng l vn qi khng th thiu c i vi mt doanh

nghip nng ng, v vy Vedan lun tuyn dng v to c hi cho mi nhn vin ca mnh theo ui mc tiu v pht trin ngh nghip. Tiu chun tuyn dng ca Vedan l nhng nhn vin c o c v kinh nghim cng tc. Trong mi trng cnh tranh x nghip mang tnh ton cu hin nay, Vedan khng ngng vn ln, hon ton nh vo k hoch o to pht trin ngun nhn lc, tng cng qun l, dn dt cng ty ra sc phn u thnh cng bc vo th k mi. n vi Vedan, cc nhn vin s c c hi tr ht ti nng, nim am m cng vic trong nghin cu sng to, v c lm vic trong mi trng nng ng. Ngoi ra cc nhn vin cn c hng nhng phc li x hi khc, nhm nng cao tinh thn hng hi v kch thch nng lc t pht ca bn thn.
1.7.

Vn ha cng ty Ngy nay, vn ha cng s c xem nh l yu t cn bn to nn s khc

bit ca mt thng hiu. cc gi tr ny c hnh thnh t tnh cch, mong

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mun ca ngi lnh o, ng gp ca ton th nhn vin trong qu trnh sn xut kinh doanh. Lun tch ly, iu chnh theo thi gian cng vi s pht trin ca cng ty. Mi trng vn ha Vedan c xy dng phong ph, vi nhiu bn sc, gn kt cc c nhn trong mt tp th nhm xy dng mi trng pht trin mnh m, bn b v l gi tr pht trin trng tn ca mt t chc. n to nn mt hnh nh khc bit ca Vedan. Vn ha th hin qua: Cch ng x hng ngy ca mi cng nhn vin. Gia cp qun l vi nhn vin ca mnh, v gia nhn vin vi nhau. Vn ha sinh hot hng ngy nh n ung, giao tip, cng vic Tt c cc yu t vn ha trn c hnh thnh v pht trin theo s ln mnh ca Vedan. N to nn hnh nh tt p ca Vedan trong mt moi ngi. 1.8. Chnh sch mi trng Bo v mi trng v pht trin bn vng l hai yu t khng th tch ri nhau i vi qu trnh pht trin ca mt nn kinh t - x hi hin i. V vy trong qu trnh kinh doanh lu di ca mnh, Vedan lun n lc duy tr v thc hin lin tc. Vedan lun c gng phn u tr thnh doanh nghip pht trin bn vng gn lin vi bo v mi trng trong thi gian ti, cng l mc tiu lu di ca cng ty CPHH VEDAN VIET NAM.

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Chnh sch h thng qun l mi trng ISO 14001 : 2004 c Vedan ly lm nn tng thit lp nn h thng qun l mi trng, ng thi nh

hng chung cho tan th nhn vin ca cng ty VeDan lun cao khi nim v yu qu mi trng. Vedan cng nh hng cho nhn vin ca mnh cao quan nim An ton v sc khe ngh nghip, phng pht sinh

cc ri ro m gy ra nguy hi cho nhn vin, tn tht ti sn v thit hi cho mi trng lm vic, thng qua Chnh Sch An Ton V Sc Khe Ngh Nghip OHSAS 18001 :2007 P Cng Ty An Ton, Mi Ngi Khe Mnh.
1.9.

Vinh d cng ty Vi nhng n lc phn u v pht trin khng ngng ca tp th cn b

cng nhn Vedan. Nm 1999, cng ty Vedan c chng nhn t tiu chun ISO 9002 v c ch tch nc tng thng Hun chng lao ng hng 3 vo ngy 08/11/2000 v nhng ng gp xut xc ca cng ty Vit Nam.

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B thng mi nc CHXHCN Vit Nam tng thng bng khen cho cng ty vo ngy 19/07/2001 v thnh tch xut xc trong nhp khu. Vi nhng thnh tu t c ca mnh, ngy 11/10/2009, Cng ty CPHH Vedan c trao gii thng Doanh nghip ng dng h thng qun l xut sc v Top 100 sn phm v an ton sc khe cng ng.
1.10.

C cu t chc hnh chnh

T chc gi nhng chc nng chung l qun l cc hot ca cc b phn trong t chc, bao gm cc khi, cc cp, vn phng hay xng (tc l quan h hng dc) m nhn nhng hot ng cn thit, xc lp cc mi quan h v nhim v quyn hn v trch nhim gia cc b phn . Mc tiu ca chc nng t chc l to nn mt mi trng ni b thun li cho mi c nhn, mi b phn pht huy c nng lc v nhit tnh ca mnh, ng gp tt nht vo vic hon thnh mc tiu ca t chc. Mi mt b phn trong c cu t chc ng mt chc nng ring v chu s gim st ca cp trn mnh. S c cu t chc:

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i hc M TP.HCM VP. TNG GIM C

KHI QUN L HNH CHNH

KHI QUN L TI V

TNG GIM C
KHI QUN L KINH DOANH

KHI M BO CHT LNG & NGHIN CU PHT TRIN KHI QUN L SN XUT

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X.LN M BT NGT EN

X.THU HI BT NGT

X.TINH CH BT NGT

X.NG GI BT NGT

P.K HOCH SN XUT

X.LN M LYSINE EN

X.THU HI V TINH CH LYSINE

X.GIA CNG SN PHMPH

X.PGA

X.XL N THI C

K Q L S X H I U N N U T
X.NHIT IN

P.C KH

P.TH NGHIM

X.ST -AXIT

X.TH NGHIM C

X.TINH BT PH THI C

P.CNG V

X.TINH BT PH LONG C

X.TINH BT BNH THUN

X.TINH BT H TNH

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i hc M TP.HCM P.THU QUAN V XNK

P.PHN TCH KINH DOANH

KHI QUN L TI V

P.VI TNH

P.TI V

P.K TON

P.VN TI

P.PHC LI NHN VIN

KHI QUN L HNH CHNH

P.T CHC HNH CHNH

P.THU MUA

P.QUN L KHO

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i hc M TP.HCM P.HA NGHIM TRUNG TM

KHI M BO CHT LNG NGHIN CU PHT TRIN

P.NGHIN CU PHT TRIN

P.M BO PHT TRIN

P.BO V

BAN QUN L NGUYN LIU

BAN D N

P.KIM TON NI B VP.TNG GIM C P.PHP CH

BAN CNG TC I NGOI

BAN TH K

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CHNG 2: TNG QUAN PHNG PHP NUI CY LYSINE


2.1. Khi nim Lysine

Hnh 2.1.a cu trc khng gian L-Lysine

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Tn quc t: 2,6 diminohexanoic acid Cng thc ha hc : C6H14N2O2 Khi lng phn t gam : 146.188 g/mol Cng thc cu to :
NH2-CH2-CH2-CH2-CH2-CH-COOH NH2

Lysine l mt a-amino acid. L acid amin thuc h aspartat, c cha 2 nhm (NH2) v mt nhm (-COOH). Lysine tng hp qua con ng trao i cht phn nhnh, qua con ng ny cn c metionin, treonin, izoloxin cng c to thnh. 2.2. Cc phng php tng hp Lysine Phng php thy phn Phng php tng hp ha hc Phng php kt hp Phng php tng hp axit amin bng cng ngh vi sinh vt : Thu nhn axit amin dng L Nng xut cao Gi thnh sn phm thp Nguyn liu sn xut r, d kim. 2.3. Tng quan v phng php nui cy Lysine 2.3.1. Ging vi sinh vt Trong nui cy Lysine. Ngi ta thng s dng chng Corynebacterium glutacium v: C kh nng tng hp Lysine vi mc cao nht, y ngi ta s dng vi khun Corynebacterium glutamicum t bin.
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Sng c trn mi trng cha hm lng methionine cao v threonine thp. Tng i d nui v p dng vo cng ngh ln men quy m cng nghip. c im ca ging: Hnh thi khun lc: trn, trn, bng Mu sc: vng chanh 2.3.2. Nguyn liu

Ngun C:

R ng ( ma hoc c ci ng). ng ca r ng bao gm: 25-40% sacaroza, 15-25% ng kh ( glucoza v fructoza), 3-5% ng khng ln men c. Cc ging vi khun tham gia tng hp Lysine u c kh nng ng ha glucose, fructose, maltose, pentose, rafinose.

Syrup glucose: dung dch ng thu c sau qu trnh thy phn tinh bt sn. CSL: bt ng c hm lng ng khong 10-20%. NH4Cl,

Ngun Nit: Pepton. Dng cc loi mui cha NH4+ nh:

NH4OH, (NH4)2SO4, NH4HPO4 hay kh NH3 hoc ure lm ngun cung cp Nit. Trong nui cy cng nghip thng s dng NH3 di dng lng hoc kh. C th s dng urue, khi s dng urue cn quan tm ti nng ban u v kh nng chu ng ca mi loi sinh vt khc nhau.

Mui khong:

Mui khong c s dng nhiu nht l cc dng

mui photpho. Nng thch hp l 0.008-0.02mg/l, trong sn xut ngi ta phi b xung thm MgSO4.7H2O vi hm lng 0.03-0.5%. Vitamin: Biotin, Nicotinic, pantothenic acid, thiamine HCL Cc ch tiu qun l cht lng mt r ( t indo, thailan, Vietnam, c ci):
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i hc M TP.HCM

2.3.3. Cc yu t nh hng 2.3.3.1. Oxy: Ln men L-lysine l mt qu trnh ln men hiu kh, i hi mt lng ln oxy. C. glutamicum c th chu c lng oxy cung cp hn ch nhng qu trnh sn xut L-lysine c thun li th cn trnh s thiu oxy bi v hot ng ca vi sinh vt s tng cng chuyn ha to ra cc ph phm, lm gim s tinh khit v hiu sut chuyn i. Trong iu kin thiu oxy, pyruvate s chuyn ha thnh cc sn phm ph nh acetate, lactate, and L-alanine [21]. Vic cung cp oxy c thc hin v kim sot bng hai cch: sc kh v khuy o. Trong khng kh, ngoi cc thnh phn ca khng kh cn cha rt nhiu VSV, do khng kh cung cp cho qu trnh ln men i hi phi c sch s v vi sinh vt v cc tp cht khc. Trng hp ln men trong iu kin hiu kh, cnh khuy lm tng kh nng ha tan ca oxy. Cc kh s li lu hn do dng chuyn ng ca mi trng, v nh vy, kh nng ha tan ca oxy t bt kh s cao hn. Ty theo qui m ln men v ging vi sinh vt s dng m tc sc kh v khuy o s khc nhau. i vi chng C. glutamicum, mt s nghin cu ln men L-lysine c tin hnh bng cch lc trong my lc (250-300v/ph) hoc bng s c kh (0,5-1,5vvm) tro n g jar ferm en tor 30 lt [28 ]. Thc hin ln men th nghim trong bench scale fermentor

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6.6 lt tc thi khng k h sch l l ,2 vvm v thay i tc khuy trong sut qu trnh ln men t 600 rpm 0 h n 900 rpm 19h [20]. 2.3.3.2. pH: pH l yu t rt quan trng, nh hng mnh m n qu trnh ln men. Nhng hp cht c bn nh sodium hydroxide, potassium hydroxide, ammonium hydroxide, calciumcarbonate, urea, ammonia v kh ammonia hoc nhng hp cht acid v c nh acid sunfuric hoc phosphoric v nhng hp cht acid hu c c s dng trong cc nghin cu kim sot pH trong ln men L-lysine dy rng pH t 5-9 [30]. pH ti u cho qu trnh sinh tng hp lysine i vi chng C. glutamicum l t 7,0 7,6 [2]. Trong qu trnh ln men, vi sinh vt li to ra nhng sn phm trao i cht c tnh acid hay kim, khin pH ca mi trng khng cn thch hp cho hot ng sng ca chng. V vy, vic ch ng iu chnh pH ca mi trng lun gi tr thch hp trong sut qu trnh ln men l iu rt cn thit. 2.3.3.3. Nhit : Nhit ti u ca Corynebacteria sn xut cc amino acid l t 25 n 37C [21]. C. glutamicum c nhit ti u l khong 30oC, nhit ti u cho qu trnh ln men lysine duy tr trong khong t 28-30oC. Do vic s dng C. glutamicum cc nc nhit i c ngha v kinh t, bi v chi ph cho vic gi nhit ti u cho qu trnh ln men khng ng k [22],[2]. Qu trnh ln men lun c s ta nhit rt mnh nn nhit trong cc thit b ln men thng c xu hng gia tng vt qu ngng nhit thch hp i vi hot ng sng ca vi sinh vt . V vy phi thng

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xuyn gim st v iu chnh nhit theo yu cu ca qu trnh ln men. Mun t c nhit thch hp cho qu trnh ln men, ngi ta trang b h thng lm nng v lm ngui bng nc chy quanh ni di dng cc ng rut g . 2.3.3.4. Hin tng to bt: Trong ln men fed-batch, hin tng to bt cng l mt iu quan trng cn nghin cu, c bit trong quy m cng nghip, khi tc kh trn b mt tng theo kch thit b. Kch thc fermentor khng phi l tiu chun nh gi tt cho qui m ca mt qui trnh cng ngh ln men. Thng thng, lng canh trng ln men trong bnh phn ng chu nh hng bi hin tng si bt, do trong sn xut cng nghip, cha y t 70 n 85% th tch. Hin tng to bt b nh hng bi t trng ca canh trng ln men, cng nh ph thuc vo chng sn xut, thnh phn mi trng v iu kin nui cy. Bn cnh p lc thy tnh ln trong cc thit b qui m ln do lng CO2 rt ln sinh ra trong cht lng, s si bt cng nh hng rt ln trong sut qu trnh nui cy v trong sut qu trnh downstream sau ny[21]. Hin tng to bt xut hin trong sut qu trnh ln men c th c kim sot bng cht chng to bt nh acid bo, ester polyglycol, hoc silicon v polypropylene [30].

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2.4. C ch tng hp Lysine Dextrose Piruvat

Oxalatetat -Aspartat-photphat Aspactat--sinial dehyd

Lysine Methionin

Homogerium Treonin

T s ta nhn thy, mun vi khun to ra nhiu L-Lysine th s tin ha phi theo nhnh . y s dng chng t bin mt Enzylme homoserin dehydrogenase, do s khng to thnh tronin v methionin. Kt qu LLysine c tng hp. S c ch c ch enzyme homoserine dehydrogenase

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Trong : AK = aspartokinase ; HSD=homoserine dehydrogenase ; Aspartokinase l mt loi enzyme ch b c ch khi hm lng ca c lysine v threonine tit ra vt mc. Qua s chuyn ha trn ta rt ra mt kt lun l : mun iu chnh s trn nhm mc ch ch sn xut Lysine v t c hm lng cao th cn c ch c enzyme aspartokinase v enzyme homoserine dehydrogenase . Vy vi sinh vt m ta chn lm ging ln men ( tt nhin l phi c kh nng tng hp lysine) phi sng trn mi trng c cha hm lng methionine cao v threonine thp .

CHNG 3: THC NGHIM NUI CY LYSINE CNG TY CPHH VEDAN VIETNAM


3.1. Mc ch th nghim nui cy Th nghim c thc hin trn mt loi vi sinh vt t bin, nhm kim tra nhng iu kin nui cy ti u nht p dng vo quy m nui cy cng nghip hiu qu hn. Ly th nghim trc lm tin cho th nghim sau Trong qu trnh nui cy, mi trng dinh dng c thay i nhng cc iu kin khc vn tun theo nhng yu cu v k thut. V vy chng ta c th so snh c mi trng nui cy pht trin ti u cho vi sinh vt v mang li gi tr kinh t cao hn. T xy dng c biu sinh trng nhng iu kin khc nhau.

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3.2. S khi quy trnh sn xut v thuyt minh quy trnh 3.2.1. S khi quy trnh sn xut L- Lysine
Chun b bn D Chun b bn F

St trng bn khng

St trng bn khng

Lm ngui

Lm ngui

Nguyn liu

iu ch mi trng

Nguyn liu

iu ch mi trng

St trng mi trng

St trng mi trng

Vitamin, H3PO4

Lm ngui

Vitamin, H3PO4

Lm ngui

Ging

Tip ging

Chuyn bn

Nhn ging

Nui dng

FD

B.O

3.2.2. Thuyt minh quy trnh 3.2.2.1. chun b bn khng Cu to chung v chc nng: Bn D:

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1: ng ng kh thi ng ng B.O 2: Thit b thao tc tip ging 3: Bn tip ging 4: ng ng bm Vitamin 5: Trc khuy 6: Cnh khuy

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7: ng ng nc vo 8: ng ng nc ra 9: ng ng gi vo 10: ng ng hi nng 11: Bn bng 12: ng ng NH3 13: ng ng D-F, ng ng ly mu.

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Bn F:

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1: ng ng B.O 2: ng ng bm Vitamin 3: ng ng D-F 4: ng ng kh thi 5: ng ng gi vo 6: Trc khuy 7: ng ng nc ra


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8: Cnh khuy 9: ng ng cool 10: ng ng hi nng 11: ng ng kh 12: Bn bng 13: ng ng NH3 14: ng ng nc vo 15: ng ng ly mu

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Chc nng: ng dn ng FD: dn mi trng dinh dng b xung trong qua trnh nui cy. Vch khuy ( vch cn dng): c tc dng nh ri mi trng nui cy.

ng gi: gm van mt chiu v thit b lc gi, ti y khng kh c thi vo cng NH3 Trong qu trnh nui cy nhm cung cp oxi v nit cho qu trnh ln men hiu kh, ng thi gip n nh PH cho mi trng nui cy. Kh cung cp vo phi sch v vi sinh vt v cc tp cht khc.

ng tip ging ng ly mu kim tra Cnh khuy: Thc hin qu trnh khuy o nhm:

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Lm tng kh nng tip xc gia cht dinh dng v t bo vi sinh vt, t lm tng kh nng trao i cht. kh nng tip xc cng nhiu th trao i cht cng mnh. Trong khi tin hnh trao i cht, cc cht sau ng ha v d ha s to ra mt lp quanh t bo. Khi cnh khuy hot ng, lp bao quanh ny b ph b. v vy mc xm nhp ca cht dinh dng vo t bo mnh hn. Ln men trong iu kin hiu kh, cnh khuy lm tng kh nng ha tan ca oxy, cc kh s li lu hn do dng chuyn ng ca mi trng. nh vy, kh nng ha tan ca oxy t bt kh s cao hn
Cnh khuy lm tng kh nng tch cc kh CO2, H2S, NH3

sinh ra t qu trnh trao i cht, lm gim nh hng xu ca cc lai kh ny n vi sinh vt. Cnh khuy lm tng nhanh qu trnh sn xut ca con ging, do tc ng c hc m cc t bo d dng tch ra v sng c lp.

ng dn kh CO2: gip kim sot c s to thnh CO2 trong qu trnh nui cy. Lng vi sinh vt cng cao th CO2 thi ra cng nhiu.

Mn hnh my tnh: hin th cc thng s k thut ca bn ln men, gip kim sot c mi trng nui cy theo ng yu cu k thut.

3.2.2.2. St trng bn khng


Mc ch: qu trnh chun b cho iu ch mi trng. Cc bin i:

Vt l: 1210C +/-20C, 1kg/cm2 ( thng s c th thay i ph thuc vo iu kin thit b). Ha hc: khng xy ra.
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Sinh hc: Tiu dit ton b vi sinh vt c trong bn cha.


Thao tc thc hin:.

M nh van y bn, van ly mu, van thi ca bnh tip ging, van ng h p sut NH3, van gi ra. M hi nng t t tin hnh st trng, khi nhit n 100 oC, ta iu chnh li van thi cho va thot hi. Khng ch nhit v thi gian theo yu cu ca Bng k hoch thc hin. Sau khi st trng xong ta m van gi vo trc, sau mi ng li van hi nng, ng thi m ln van thi thi kh bn.

3.2.2.3. iu ch mi trng
Nguyn liu bnh C:

1. Glucose 2. Fructose 3. Acid citric 4. NaCl 5. Yeast Extract 6. Circle Grow


7. CaCO3

Nguyn liu bn D:

1. Sucrose 2. Peton 3. Acid citric


4. (NH4)2SO4 5. MgSO4.7H2O 6. KH2PO4 7. Na2HPO4.12H2O 8. MnSO4.H2O Sinh vin: Bi Trung Tn -27Lp: SH08-A2

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9. FeSO4.7H2O 10. CuSO4.5H2O 11. ZnSO4.7H2O 12. Cht kh bt EP-530

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13. CSL_Requette 14. Biotin 15. Ca_pantothenate 16. Nicotinic acid 17. Thiamine HCl
Nguyn liu bn FD:

1. Dextrose 2. Mol INDO


3. Cht kh bt EP-530

4. Biotin
5. (NH4)2SO4. c ha tan v st trng ring vi cc nguyn liu khc

121+/-20C, 1kg/cm2, thi gian l 25 pht ( ty thuc vo yu cu thc nghim). Sau c trn chung vi nhau.
Nguyn liu bn F:

1. Sucrose
2. (NH4)2SO4 3. Cht kh bt EP-530 4. Na2HPO4.12H2O 5. KH2PO4

6. Acid citric
7. MgSO4.7H2O 8. FeSO4.7H2O 9. CuSO4.5H2O 10. ZnSO4.7H2O

11. CSL_Requette

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12. Biotin
13. H3PO4

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14. Ca_pantothenate 15. Nicotinic acid 16. Thiamine HCl

Thao tc thc hin: ng van gi v trng, van y bn, van ly mu, m van thi p sut v 0 kg/cm2. M np bn ra cho nc vo trc, ri mi cho cc loi nguyn liu theo Bng pha ch mi trng, sau dng nc nh mc theo yu cu. Sau khi cho nguyn liu vo, khi ng motor khuy v ly mu o Bx, pH. ng np bn li dng NH3 iu chnh pH theo yu cu ca thc nghim. Mi trng c iu ch v thay th nguyn liu ty theo yu cu thc nghim. Hiu chnh PH t 6.8 7.2 bng NH3 ty theo c tnh ca tng loi ging. Nhm p ng cho nhu cu sinh trng, pht trin ca sinh vt.

3.2.2.4. St trng mi trng Mc ch: L qu trnh chun b cho qu trnh nhn ging v qu trnh nui dng. Cc qu trnh bin i xy ra: Vt l: 121+/-20C, 1 kg/cm2 ( thng s c th thay i ph thuc vo iu kin thit b). Ha hc: ng nht mi trng. Sinh hc: Tiu dit ton b vi sinh vt c trong bn cha mi trng.

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Thao tc thc hin:

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Thi gian st trng ty thuc vo yu cu thc nghim. St trng c chia lm 3 giai on: nng nhit, gi nhit, lm ngui. M hi nng t t ra. M ln ht van tip nhn ging v van chuyn tip ging, van thi kh ch m mt na, m 1/3 van y bn v van hi nng ca ng ly mu. Khi nhit ln ti 100oC , t van y bn v van ly mu x ra mt t dch mi trng trnh tnh trng st trng khng trit . Nhit v thi gian st trng tun th theo qui nh trong Bng k hoch thc hin. Sau khi hon tt st trng, x van y bn v van ly mu mt t dch mi trng trnh tnh trng st trng khng trit sau ng li van y bn v van ly mu li ri mi cho thng hi nng phng b nhim tp khun. Sau qu trnh lm ngui, Vitamin c bm vo bn cho ging vi sinh vt s dng. ng li van hi nng, van tip nhn ging v van chuyn tip ging.
Thao tc lm ngui mi trng:

Sau khi hon tt vic st trng. M van gi trc, sau ng van hi nng li. ng van x nc, sau m nc lm mt h nhit 35-37oC. Sau khi lm ngui mi trng, dng NH3 chnh pH mi trng ln theo yu cu th nghim v ch chuyn bn. Trc ht phi thng gi bo p mi m nc lm mt, trnh thn bn b bin dng. 3.2.2.5. Ging v tip ging Ging c phn lp, chn lc v c nhn ging cp 1 trong ng nghim ( ng B), nhn ging cp 2 trong bnh tam gic ( bnh C) cha dch nui cy. Lc ging cp 2 trong bnh tam gic t 13-16h ( ty tng loi ging). Tip ging:

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Tt c cc thng s k thut trong bn nui cy phi t yu

cu: gi, p sut, nhit , PH Ging c tip t bnh tam gic vo bn nhn ging (bn D) bng bm lu lng. Tin hnh cy tp khun t ging cp 1, cp 2 trc ( trc khi tip ging) v dch ln men sau khi tip ging. Nu trn mi trng dinh dng nui cy tp khun, dch c cy vo c pht hin khun lc. Chng t ging nhim tp khun.

3.2.2.6. Nhn ging


Mc ch: Tng sinh khi vi sinh vt

Cc qu trnh bin i xy ra: Vt l: Qu trnh trao i cht ca vi sinh vt lm nhit trong bn tng ln. Ha hc: Lm gim pH mi trng do to ra acid trong qu trnh chuyn ha ca vi sinh vt. Sinh hc: ging c bm vo s dng c cht c iu ch sn tng sinh khi. iu kin nhn ging: Nhit p sut pH Thi gian
Thao tc thc hin: kim sot cc thng s: pH, p sut, nhit , lu

lng gi. Theo di CO2, OD, OUR khi t yu cu theo bng k hoch thc hin E100040 th chuyn bn.
Thit b: bn D

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3.2.2.7. Chuyn bn

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Khi bn D t yu cu ca Bng k hoch thc hin th chuyn t bn D sang bn F.

3.2.2.8. Nui dng

Mc ch: Ln men to Lysine. Cc qu trnh bin i xy ra: Vt l: Nhit tng dn. Ha hc: pH mi trng gim trong qu trnh chuyn ha ca vi sinh vt do to ra nhiu acid. To ra lysine v cc sn phm khc. Sinh hc: Qu trnh ln men c thc hin trong thit b c cnh khuy, v thi kh lin tc v vi sinh vt c nui cy trong mi trng dch th, chng pht trin theo chiu ng ca ct mi trng. V vy, phi thc hin yu cu v khuy o v sc kh. Sau khi ging c tip vo, tin hnh nui cy t 68-72 gi. Qu trnh ln men, di s khuy o lin tc trong bn ln men s c hin tng xm thc ( s va p ca dch ln men vo thit b kim loi), v qu trnh sinh kh ca vi sinh vt xut hin nhiu bt kh c khuynh hng tro ra khi bn ln men, d gy tp nhim. V vy, cn kim sot lng bt v tm cch ph hy chng. Ngoi ra, bt kh cng cn tr s tip xc ca t bo vi mi trng dinh dng. L-Lysine c cu thnh t cc phn t C, H,O,N. V vy, mi trng nui cy phi p ng cc yu t cn thit hnh thnh cu trc vi sinh vt.

iu kin ln men Nhit , pH, gi da theo Bng k hoch thc hin.

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Cc cht dinh dng iu ch theo Bng pha ch mi trng.

Thao tc thc hin: Kim sot: pH, p sut, lu lng gi, nhit . Theo di OD v tin hnh FD theo Bng k hoch thc hin.
Thit b: bn F.

3.2.2.9. B.O Qu trnh nui dng kt thc trong khong thi gian 68 72 gi. B.O khi pH ln. Thi gian kt thc qu trnh nui dng da vo : o Da vo ng cong sinh trng nui cy vi sinh vt c thit lp trong cc th nghim.
o

Da vo kt qu phn tch TS, MV, OD ca ln phn tch cui xc nh thi im B.O.

Thu nhn sn phm, tin hnh ly mu phn tch v kim tra pH, TS, OD, MV. Ty thuc vo nhu cu nui cy m sn phm c sy kh, tinh ch , phc v nhu cu sn xut khc hoc kh trng x b.

Lng t bo Pha cn bng Pha ly tha Pha tim n pha suy vong

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68h

72h thi gian

Hnh 3.4.e ng cong sinh trng nui cy vi sinh Thi im thu nhn sn phm c th giai on cui ca pha ly tha hoc u hoc gia pha cn bng. Tuy nhin, nn B.O ti thi im cui pha ly tha v pha cn bng ( t l vi sinh vt sinh ra bng vi t l sinh vt cht), s lng vi sinh vt cht c th gy nh hng ti cht lng ca sn phm.

3.3. Phn tch v kim tra: Trong qu trnh ln men, mi trng nui cy s c s thay i do nhng phn ng xy ra lm thay i mi trng. v vy, phi kim tra theo nh k v cc thng s nh: T/S, OD, MV, PH t 1- 4 gi/ 1 ln. PH: mi loi vi sinh vt thch nghi vi mt mi trng nui cy. trong qu trnh ln men, vi sinh vt to ra nhng sn phm trao i cht c tnh acid hay kim, khin pH ca mi trng khng cn thch hp vi mi trng sng ca mnh. V vy, cn phi thng xuyn theo di v iu chnh pH trong sut qu trnh nui cy. Vi khun Corynebacterium glutamicum sinh trng ti u pH t 6.8 7.2. V vy trong qu trnh nui cy, mi trng c nh hng nhiu ti nng sut Lysine.
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Nhit : nhit l yu t quan trng nh hng ti s pht

trin ca vi sinh vt v hiu qu ca qu trnh ln men. Mi loi vi sinh vt thch ng vi mt nhit khc nhau ( y nhit t 32 340C). Trong qu trnh ln men lun c s ta nhit rt mnh, nn nhit trong cc thit b ln men thng c xu hng gia tng vt ngng nhit thch hp i vi hot ng sng ca vi sinh vt . V vy, phi thng xuyn theo di v iu chnh nhit thch hp cho qu trnh ln men ( 32 340C). t c nhit thch hp cho qu trnh ln men. Bn ln men c trang b h thng lm nng v lm ngui bng nc chy quanh y bn di dng ng rut g lm ngui.

T nghin cu l thuyt n thc nghim ta thy, nhit mi trng t 30 400C lng lysine sinh ra cao hn, c bit nhit t 32 340C. V vy trong sn xut v th nghim chng ta chn nhit ny ( 32 340C).

Phn tch T/S: Mc ch: Kim tra hm lng ng v iu chnh ph hp. Ha cht s dng: dch A, dch B, dch C,dch D. Tin hnh: ly mu t bn nui cy Ht 10ml dch mu cn phn tch Ht 1ml hoc 2ml H2SO4 30% ( ty thuc vo tui ca mu): Mu th t 20 40 tui dng 1ml H2SO4 30% Mu th t 40 72 tui dng 2ml H2SO4 30%

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Cho vo ng nghim v nh mc bng pH=1 ti 30ml. Tin hnh gia nhit trong 30 phut thy phn hon ton hm

lng ng trong dch mu. sau khi gia nhit, tin hnh lm ngui v iu chnh pH ca mu t t 6,5 7,3. nh mc ti 100 ml bng nc siu thun. Cho vo bnh tam gic (250 ml ) 20 ml dch A. Ht 2ml hoc 5ml dch mu ( nu T/S ln kim tra trc t 3.00 tr ln th ht 2ml, nu di 3.00 th ht 5ml), cho vo dch A, un si trn bp 3 pht v lm ngui. Thm vo 10ml dch B, 10ml dch C. Tin hnh chun bng dch D ti khi i mu, thm 3 git ch th vo v tip tc chun ti khi i mu hon ton. Da vo th tch dch D, xc nh T/S theo cng thc: %TS = ( Vo V1)xFx1.449 x 0.2 Hoc %TS = ( Vo V1)xFx1.449 x 0.5 Vo: s ml chun mu trng V1: s ml chun mu th F: h s dung dch D. o MV: Xc nh vi sinh vt v sinh khi ca vi sinh vt dng th. Tin hnh: cn c theo MV ca ln kim tra trc hoc mu kim tra m tin hnh o MV Nu MV ca ln kim tra trc cao hoc dch qu c, kh c th nhn bit c th tin hnh pha long mu. ht 10 ml dch mu , ly tm trong 10 pht ( 3000 vng/ pht). o OD: Xc nh c ca mu, t xc nh mt vi sinh vt c trong dch mu.

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Ht 1ml dch mu, nh mc ti 100ml v o mu bng my o

quang bc sng 660nm.

CHNG 4 : GII THIU V VAI TR V NG DNG SN PHM


4.1. Vai tr v ng dng:
Lysine gi vai tr sng cn trong tng hp protein, l cha kha trong sn xut enzyme, hoocmon v cc khng th gip c th tng cng sc khng, chng bnh tt c bit ngn cn s pht trin ca vi khun gy bnh mn gp mi hay mn gp sinh dc. C th ngi v ng vt thiu lysine c th s khng hot ng bnh thng, c bit ng vt cn non v tr em s xy ra hin tng chm ln, tr tu pht trin km. Chnh v th lysine thng c a vo phn n ca tr v ca gia sc. L-lysine l mt amino acid cn thit v i hi phi lun c sn trong thc n p ng nhu cu dinh dng ca ng vt, c bit i vi thc n t ng cc, la m hoc la mch th ngho lysine. Do b sung ngun giu lysine vo l cn thit tng hiu qu thc n. Ta c th b sung bt u nnh (cha nhiu lysine) hoc thm trc tip lysine vo thc n. u im ca vic b sung trc tip lysine l cc amino acid khng thay th khc khng c thm vo nn thnh phn ca chng trong c th khng b thay i. V d thm 0.5% lysine tng cht lng m ca thc n hiu qu nh l b sung 20% u nnh. T , nit ca amino acid khng gii hn thm vo tha ca ch n c hm lng m cao th s b chuyn ha thnh amoni v c ng vt bi tit ra ngoi, vi vic b sung lysine lm gim lng m b sung t lm gim nhim mi trng t phn.

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Hnh 4.1.1. Nhu cu ca lysine, threonine v methionine trong thc n heo con v thnh phn ca nhng amino acid ny cha sn trong thc vt [34]. Trong nm 2000, vic sn xut L-lysine khp ni trn th gii c s dng nh l mt ngun cht b sung vo thc n t sp x 550.000 tn v trn th trng vn cho thy mt s tng trng tim nng t 7-10% mi nm [34]. V ch c L-lysine l c hiu qu nh l mt ngun cht b sung v tt c cc quy trnh sn xut hin nay s dng phng php ln men quen thuc.

Hnh 4.1.2. Sn xut lysine ln men (tn/nm) trong sut 3 thp k qua [34]. Lysine c nhiu trong trng, tht, c, u nnhnhng d b ph hy Sinh vin: Bi Trung Tn -38Lp: SH08-A2

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trong qu trnh ch bin v nu nng thc n. Ngi ta b sung lysine cho ng vt di dng thc n nh cho lysine vo sa. ngi lysine c b sung vo thuc dng vin v thuc ung.

4.2. Cc dng tn ti ca lysine:


L-lysine Sulphate

Hnh 4.2.1. L-lysine sulphate [18]. Thnh phn: Lysine Sulphate l mui sulphate vi lysine L-lysine : 51% min, trung bnh l 65%.. Sulphate : 15% max Cc acid amin : 10% min. m : 3% max Mui amoni (nh NH4) : 1% ma Kim loi nng : 0,003% max Arsen : 0,0002% max. Lysine Sulphate c th c b sung vo L-lysine HCl lm thc n cho vt nui, chng c hiu qu nh L-lysine HCl. L-lysine HCl (L-Lysine monohydrochoride) [18]. Xut x : Trung Quc

Hnh thc : Mu nu sng.

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Hnh 4.2.2. L-lysine HCl [18]. Thnh phn : 98.5% m : 1% Amonium (NH4) : 0.04% Kim loi nng : 0.003% pH (1 :10 nc) : 5 - 6.09 Hnh 2.5: L-lysine HCl L-lysine HCL 98.5% (Feed Grade) l mt trong nhng amino acid c s dng rng ri nht. y l amino acid cn thit yu cu c trong ch n ca ln, g vt gia cm.

4.3. Cc sn phm lysine thng mi:


L-lysine vi B6 [37].

Hnh 4.3.1. Sn phm thng mi L-lysine vi B6 [37]

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Thnh phn: L-lysine, vitamin B6 (Pyridoxine hydrochloride), Mg, Si, men, gluten, mui, ng v cc sn phm sa. Mi Capsule s cung cp: L-lysine 500mg, Vitamin B6 6mg Gi thnh: 380000 VND. Lysine power [40].

Thnh phn ca sn phm ny l L-lysine HCl.

Hnh 4.3.2. Lysine capsures [40].

Hnh 4.3.3. Sn phm thng mi Lysine Capsures. Thnh phn ca sn phm ny l L-lysine HCl [40]. Lysine Vit- B12 Granules [40].

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Hnh 4.3.4. Sn phm thng mi Lysine Vit B12 Granules [40]. Thnh phn: 300mg lysine, 15mg vitamin B12 Granule c hng v ngt. Lysine-1000 90ct [40] Lysine ng vai tr quan trng cho s pht trin ng n v ng vai tr thit yu trong sn xut carnitine. N chu trch nhim chuyn i cc acid bo thnh nng lng v gip gim cholesterol. Lysine xut hin gip c th hp th v chuyn ha canxi, n ng mt vai tr quan trng trong vic hnh thnh collagen, mt cht quan trng cho xng v m lin kt bao gm da, gn v sn.

Hnh 4.3.5. Sn phm thng mi Lysine-1000 90ct [40]. Lysine Powder [40]

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Hnh 4.3.6. Sn phm thng mi Lysine Powder [40]. Thnh phn L-Lysine

CHNG 5 KT LUN V NGH 5.1. Kt lun 5.2. Kin ngh TI LIU THAM KHO

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