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rӗi FkQ7tQK ӕL ӧng Oѭ WRiQNK sinh khӕL NK{WK{QJ FKrQK Ӌ ch khӕ O LOѭ ӧng [19] 2.3.3 3KѭѫQJ SKiS ҧ P ÿiQK TXDQ JLi YjF TXDQ Ӌsӧ ViW i nҩ m Vz KuQK YjQJ a ĈiQKJLi ̫m quan b̹ FQJ SK˱˯QJSKiSTXDQ ̷c, tr̩QJWKiLYj ViWPjX ͡ Gj\F ͯa h͏sͫi [3] + HӋsӧL ҥ W ÿ \rX ҫ u: hӋsӧ FL PjX ҳ QJ ӗ WU ng nhҩ ÿ t, mӑc khӓe, thҷ QJ Yj FKL QKiQK Ӆ u, thuҫ ÿ n chӫ ng + HӋsӧL NK{QJ ҥ W \rX ҫ u:ÿhӋsӧ FL әL ÿ PjX Ӄ W ұ m,ÿ Y vӃ t nhҥ W ӏ FK Fy PjX G YjQJ + HӋsӧL NK{QJ Ӈ Q SKiW NK{QJ Ӌ WUL Qӧ Wѭ ng Fy bung KL sӧi, mү u chӃ t, nhiӉ m vi sinh vұ t b 4XDQViWKuQK ͏sͫi ṋm VzYjQJ WKiLK Lҩ y mӝWӧ tWV i nҩ PGjQ Ӆ XYjR ÿӑWQѭ ӟF JLFyWUrQ PNt nh Cӕÿӏ OD nh sӧi nҩ m WUrQODP bҵ QJNtQK FiFK ҽODP KѫNtQKTXD QK ҥ L WUrQ ӑQ ÿqQ O ӗ QJ n Nhuӝ Fm bҵ ng dung dӏ ch xanh methylen 5- SK~W5 ӱa thuӕc nhuӝm bҵ ng cӗQ6DXÿy ӱa lҥ i vӟLӟ Qѭ c cҩ W/rQNtQK ҧ Q TXDQ ӟ Gѭ i vұ W NtQK ViWWLrXE [6] [ 2.3.4 ;iFÿ ӏ QKWKjQKSK ҫ QGLQKGѭ ӥng quҧthӇYjWѫQ ҩ m VzYjQJ [10], [17] Sӱdө QJ FiF SKѭѫQJSKiS ӇÿiQK JLi hKyD phҫ QGLQK WKjQ VLQK ӥng cӫ aGѭ ÿ hӋ sӧi nҩ m VzYjQJ Yj ҧthӇ TX nҩ m VzYjQJ : - ;iF ӏ QK ÿӝҭ ÿ m cӫ a hӋsӧi nҩ m bҵ QJ SKѭѫQJ ҩ \ NK{ SKiS Ӄ n khӕL ÿV ӧng Oѭ NK{QJ әi ÿ - ;iFÿ ӏ QKKjP ӧng protein Oѭbҵ QJ SKѭѫQJ SKiS %LXUH 17 - ;iFÿ ӏ QKKjP ӧng lipid Oѭ bҵ QJ SKѭѫQJ SKiS 6R[KOHW - ;iFÿ ӏ nh KjP ӧngOѭ tro bҵ QJ SKѭѫQJӃ SKiS n khӕL ӧQJ Oѭ QXQJ NK{QJ әi ÿ ÿ - Ĉӏ QK ӧng Oѭ vitamin B1 bҵ QJ SKѭѫQJ Ǥ nh quang SKiS KX 2.3.5 ;iFÿ ӏ QKGѭ ӧc chҩ WFyWURQJK Ӌsӧi nҩ m, quҧthӇVzYjQJ YjG ӏ ch thӇsau QX{LF ҩ y a Chu̱n b͓m̳u th͵cͯa h͏sͫi ṋm VzYjQJ 7ѫ ҩ P Q QX{L WURQJӡ P{L ng lӓng Yj WUѭ ҧthӇ TXnҩ m Vz YjQJ thu hoҥ FK ӧcÿѭ ÿHP sҩ \ NK{ ӗL [D\ U ӝW WKjQK VDX QJkP Eӟ ÿy i nhӳ YQJ GXQJ WKtFK P{L ӧp rӗi K tiӃ QKjQK F{TXD\ Ӈthu dӏ ch ÿ chiӃ t b 3K˱˯QJ͓ QKO˱ SKiSÿ ͫng polysaccharide ¾ Ĉ͓ QKO˱ ͫng polysaccharide ngo̩LEjR WK{ d͓ FKQX{LF ̭\WƭQKYjO ̷c h͏ sͫi ṋm VzYjQJ [21] Dӏ FKQX{L cҩ \ÿѭ ӧc lӑc bҵ ng giҩ y lӑc, phҫ n qua giҩ y lӑFÿѭ ӧc thu lҥ i Giҩ y lӑc sau lӑFÿѭ ӧc sҩ y ӣ110 0C 4h rӗLÿHPFkQNK ӕLOѭ ӧng Phҫ n dӏ ch lӑFÿHP ÿLÿRWK ӇWtFKWL Ӄ SÿyWKrPO ҫ n thӇWtFKHWKDQRO YjRÿ Ӈtҥ o kӃ t tӫ D6DXÿy tiӃ QKjQKO ӑc thu tӫ a Sҩ y tӫ a ӣ45 0C 36 giӡÿѭ ӧc polysaccharide ngoҥ L EjR WK{7L Ӄ Q KjQK FkQSRO\VDFFKDULGH ҵ QJ FkQSKkQ WtFK E KhӕLOѭ ӧng polysaccharide ngoҥ L EjR ӧFWtQKWKHRF{QJWK ÿѭ ӭc sau: mpolysaccharide = msau ±mEDQ ҫ u ÿ mpolysaccharide: KhӕLOѭ ӧng polysaccharide ngoҥ L EjRWK{ J msau: KhӕL ӧng Oѭ sau sҩ y gӗm polysaccharide ngoҥ L EjRҩ WK{ y lӑc Yj (g) mEDQ ӧng giҩ y lӑc (g) ҫ u: Khӕ ÿ LOѭ ¾ Ĉ͓ QKO˱ ͫQJȕ -glucan tͳsinh kh͙i qu̫th͋YjK ͏sͫi ṋm VzYjQJ [19] &kQ PJ ӕL NK{ VLQK Yj ӟ NK c cҩ W ÿHP POQѭ ҩ p ӣnhiӋ KW ӝ121 ÿ 0& iS suҩ W DW WURQJ ӟiSK~W tӕF ӝÿ 6DX YzQJSK~W ÿy O\ WkP WU Y 18 Lӑc qua giҩ y lӑc Dӏ ch lӑF WKXÿѭ ӧc pha trӝn vӟi lҫ n ethanol 96% lҳ c mҥ QK Yj ÿӇTXDÿrP ӣ40&6DXÿyO\WkPY ӟi tӕFÿ ӝYzQJSK~WWURQ ҫ n nәL OrQ ӧc bӓÿL ÿѭ Yj ҫ n huyӅ SK Q SKӟ SKtD Lӧ ÿѭ c ӫvӟiGѭ NaOH 1M ӣ60 0C WURQJK/ѭ ӧQJ JOXFDQ ӧF[iFÿ ӏ nh ÿѭ bҵ QJ SKѭѫQJ ± SKiS sulfuric SKHQRO 3KѭѫQJSKiSSKHQRO ±VXOIXULF ӏ nh glucan [iFÿ dӵDWUrQ Ӌ c hҩ p thө YL tҥ Lӟ Eѭ c VyQJ QP ӫ a phӭc chҩ tF tҥ o bӣL SKHQRO cbohydrate Yj SӱFD dө QJ Pi\ TXDQ phәhҩ p thө , chӑQӟ Eѭ FVyQJ QP ӵQJ ӡng ÿѭ chuҭ Q [k\G JOXFDQÿR ү u SKkQ WtFK ;k\ ӵQJ G ӡng ÿѭ chuҭ n glucan: Pha dung dӏ ch chҩ t chuҭ Q Oj -glucose ' vӟi nӗQJÿ ӝ1mg/ml (1000ppm) Tӯdung dӏ ch chuҭ QWD SKD WKjQK ҭ n vӟL FiF Gm\ nӗQJÿ ӝPJPO PJPO PJPO ҩ y 5ml mӛi nӗQJ ӝchҩ ÿt chuҭ QÿѭD YjR ӑPO FiF ҫ Q 7KrP Oӧ Oѭ t tҩ t cҧO FiF ӑchuҭ On POSKHQRO VDX ҫ QOѭ ӧW YjR ҩ t cҧFiFO ÿyWKrP Wӑ2,5ml dung dӏ ch OH2 SO4 96% CuӕL FQJ ÿR ÿ{ hҩ ҕ p thөquang hӑc ӣEѭ ӟF VyQJ QP Tӯÿѭ ӡng chuҭ Q [k\ ӵng [iF Gӏ QK ÿӧcÿѭ nӗQJ ӝmү ÿX SKkQ jP WtFK ӧng Oѭ + ȕ-JOXFDQ ӧFWtQK ÿѭ WKHRF{QJWK ӭc sau: +jP ӧng Oѭ ȕ-glucan (%) = షయൈ଼ൈଵ ௫ሺ ሻ ൈଵ ெ் ሺ ሻ 7URQJ[OjV ÿy ӕJDP [iFÿ ӏ nh tӯÿѭ ӡng chuҭ n OjK ӋsӕSKD ORmQJ /037ӕ OjNK LOѭ ӧng mү X EDQ ҫ XÿHPSKkQ ÿ WtFK c ;iFÿ ͓ nh ho̩WWtQKNKiQJNKX ̱n cͯa h͏sͫi ṋm YjTX ̫th͋VzYjQJ [34] Sӱdө ng chӫ ng chuҭ n vi khuҭ n E coli, tiӃ Q KjQK ӱkhҧQăQJ WK NKiQJ ҭ n N cӫ a dӏ ch chiӃ t tӯWѫQ ҩ PYjTX ҧthӇWKHRSKѭѫQJSKiSNKX Ӄ FKWiQWUrQÿ ҥ ch, ÿӕi chӭQJNK{QJ ӱdө ng dӏ chV chiӃ t NӗQJÿ ӝchҩ t chiӃ t tҥ R UDYzQJ ҭ n ӭc Y{N chӃsӵVLQKWUѭ ӣng cӫ a vi khuҭ n 19 d Th͵ho̩W WtQK ͙ ng oxL FK KyD ͯa h͏sͫ Fi ṋm Vz YjQJ b̹QJ SK˱˯QJ SK Reducing power [21] Sӱdө QJ SKѭѫQJ SKiS Ӈ5HGXFLQJ kiӇ m tra hoҥ W WtQK ӕ SRZHU QJ R[L FK ÿ Ky Lҫ QOѭ ӧt lҩ y dӏ ch chiӃ W WKHR ӇWtFK Gm\WK VDX PO PO ӗi VDXÿy 2ml WKrP dung dӏ ch ethanol 96%, 2ml dung dӏ FK Ӌ mÿ phosphate (0,2 M; S+ YjPO NDOLIHUULF\DQLGH ӕng nghiӋ P Yj ӫӣ300 C 30 SK~W7L Ӄ Sÿy WKrPYjR ӛn hӧp ml trichloroacetic K acid 10% rӗLÿHP ӣOLWk tӕFÿ ӝ YzQJSK~WWURQJ ӕLFQJO ҩ y 3ml phҫ SK~W n dung dӏ FKWUrQ Ӆ &X E mһ t pha vӟi ml etKDQRO YjPOIHUULFFKO ӝhҩ p thөcӫ a dung dӏ FK WUrQ ӟi thiӃ t bӏUVY Vis ӣEѭ ӟF VyQJQP ӵc hiӋ Q WѭѫQJ ӵ7K vӟi W mү u kiӇ m chӭQJ 7Kt Ӌ P ÿѭ ӧ QJKL c tiӃ QKjQK ҫ n O AAI = ோ ௦ ோ ௧ 7URQJ : ÿy AAI Oj chӍsӕchӕQJR[\KyD Rsample OjP ұ W ӝquang ÿ cӫ a mү u Rcontrol OjP ұ Wÿ ӝquang mү u kiӇ m chӭng e 3K˱˯QJ͓ QKWtQKDONDORLG SKiSÿ ¾ ÈSG ͭQJQJX\rQW ̷c th͵cͯa Webb vͣLFiFKWK ͵QK˱VDX Bӝt nҩ m xay nhuyӉ n (10 ±J YjGXQJG ӏ ch H2SO4 ÿѭ ӧFFKRYjR H ÿXQQK ҽtrong giӡ Lӑc lҩ y dӏ FKÿ ӇthӱnghiӋ m vӟi thuӕc thӱMayer ¾ Thu͙c th͵ÿ͓ QKWtQKDONDORLG Pha thuӕc thӱMayer bҵ QJ FiFK KzD WDQJ +J&O ӟ PO c cҩ W Yj Qѭ WURQJ J , WURQJ ӟc cҩ t Thu hӛnPO hӧp Qѭ dung dӏ FK Qj\ ҥ L Yj O WKrP ӟc cҩ t Qѭ FKRÿ ӫ100ml NhӓYjL ӑt thuӕ JL c thӱ0D\HU Yj ӏ FK GXQJ DFLG G ORmQJ ӭa alkaloid, nӃ uFy FK Fy DONDORLG Ӂxuҩ t hiӋ n kӃ t tӫ V D PjX ҳ ng hoһ WU F YjQJ ҥ t Cҫ Q QK X Oѭê ӫ a Yu tҥ o W 20 WKjQKFyWK ӇKzDWDQWU ӣlҥ LWURQJOѭ ӧng thuӕc thӱhoһ FKzDWDQE ӣLHWKDQROF ҹ n dung dӏ ch thӱ[4] f 3K˱˯QJ͓ QKWtQKK SKiSÿ ͫp ch̭t saponin 7KtQJKL ͏ m t̩o b͕t: 6DSRQLQ ҥ o FyWtQKW bӑWQrQ ӝt ÿk\OjP nhӳng SKѭѫQJ SKiS Ӈÿӏ QKWtQKV FKtQK[iF ӵhiӋ n diӋ n cӫ a saponin ÿ 3K˱˯QJSKiSWL ͇ QKjQK KzDWDQ ӏ ch chiӃ POG WYjR ӟF POQѭ QyQJ ӑc / YjR ӝt ӕ P ng nghiӋ m 1,6FPYj ӇnguӝL ÿ WKrPQѭ ӟFFKRÿ ӫ10 ml, lҳ c mҥ nh dӑc theo chiӅ u ӕng nghiӋ P WURQJSK~W Ӈ\rQ ӕng nghiӋ P ĈTXDQ ӟp bӑWYj ViWO ÿiQK Ӄ t JLi quҧ N - Bӑt bӅ QWURQJSK~W - Bӑt bӅ QWURQJSK~W - Bӑt bӅ QWURQJSK~W +++ * Th͵nghi͏ m Fontan ±Kaudel: Lҩ y mӝWOѭ ӧng 10 ml dӏ ch chiӃ W&KLD Ӆ u2 ÿ ӕng nghiӋ m + ӔQJ WKrPPO+&O1 S+ + ӔQJ WKrPPO1D2+ 1 S+ Lҳ c mҥ nh dӑc theo chiӅ u ӕng nghiӋ P WURQJSK~W Ӈ\rQ TXDQ Yj ÿViW cӝt bӑt cҧ2 ӕng nghiӋ m + NӃ u cӝt bӑt cҧ2 ӕQJ FDRQJDQJ Ӆ QQKѭ QKDX QKDX Yj ӝ[iF EWKu ÿӏ QKOjFyVDSRQLQ WULWHUSHQRLG + NӃ X S+ FyF ӝt bӑWFDRKѫQ Ӆ u so vӟi ӕQJS+ QKL ӝVѫE [iFÿ ӏ nh OjFyVDSRQLQ [2] VWHURLG g 3K˱˯QJ͓ QKWtQKDFLGK SKiSÿͷX F˯ Lҩ y ml dӏ ch chiӃ t cӗQFKRYjR ӝt ӕng nghiӋ P P 7KrP YjR ӏ ch mӝWtW GXQJ tinh thӇNa2CO3 KѫQK ҽqua ngӑn lӱa NӃ XFyFiFE ӑWNKtV ӫ LOrQW ӯFiFWLQKW Ӈ Na2CO3 WKuN Ӄ t luұ Q OjFyDFLGK ӳX [1] Fѫ 2.3.6 ;iF ӏ nh ÿ chӍWLrXYLVLQKY ұ t hӋsӧi nҩ m VzYjQJ Ĉӏ QK ӧng Oѭ vi sinh vұ t bҵ QJ SKѭѫQJ Ӄ m sӕSKiS khuҭ n lҥ F ÿWUrQ ӡngP{L W ÿһ c Mү u cҫ Q [iF ӏ QK ӧ ÿ F ÿѭSKD ӣnӗ ORmQJ QJ ӝWKtFK ÿ ӧp T\ K YjR ӍWLrX FK YL sinh vұ t sӁFyP{LWUѭ ӡQJÿ һ c hiӋ XNKiF P{L QKDX WUѭ ӡQJ ӧcÿѭ chӭa WURQJÿƭD 21 SrWUL 0{LWUѭ ӡQJ [iF ӏ nh chӍWLrX ÿ E.coli P{LWUѭ ӡng Macconkey Agar 0{LWUѭ ӡQJ [iF ӏ nh chӍWLrXW ÿ әng vi sinh vұ t hiӃ X NKt P{LWUѭ ӡng LB (pepton: 5g, cao nҩ P PHQ J1D&O ӟc cҩ W ÿ JDJDU ӫ1l) J 0{LWUѭ ӡng [iF ӏ nh chӍ ÿ WLrXW әng nҩ P PHQYj ҩ m mӕF P{LWUѭ Q ӡng Hansen (glucose: 50g, pepton: 10g, K2HPO4 : 3g, MgSO4.7H22 DJDU ӟc cҩ tJQѭ ÿӫ 1l) 6DXNKLFy ү XSKD PORmQJYj ӡng tiӃ Q KjQKJLHR P{LWUѭ ҩ y 1ml mү u YjR F P{L WUѭ ӡQJĈHP ҩ \ ÿƭDÿm QX{L ӣÿL Ӆ u kiӋ nF nhiӋ W ӝ30 ÿ0C 48 giӡ CuӕL FQJ ÿӃ m khuҭ n lҥ c Yj WtQKN Ӄ t quҧWKHRF{QJWK ӭc sau: ܷܨܥ ܰ ܣሺ ሻൌ ݃ ݊ͳܸ݂ ͳڮܸ݂݊݅݅ 7URQJ ÿy $ sӕ Oj tӃEjR YL ҭ n (khuҭ NKX n lҥ c) 1g mү u 1Oj tәng sӕkhuҭ n lҥ Fÿ Ӄ Pÿѭ ӧFWUrQFiFÿƭDÿmFK ӑn QOj sӕOѭ ӧng ÿƭDF ҩ y tҥ Lÿ ӝSKD ORmQJ ӭi WK 9 thӇ Oj WtFKG ӏ ch mү u (ml) cҩ \ YjRӡng P{LWUѭ IOj ÿӝSKD ORmQJ ӭng WѭѫQJ 2.3.7 3KѭѫQJ QX{LWU SKiS ӗng thӱnghiӋ m a 1X{L ͛ ng ṋ WU m b̹ng gi͙ ng d̩ng l͗ ng YjG ̩ng h̩t CiFW~L3( chӭa 12JJLiWK ӇPQFѭDFDRVX ÿ ӝҭ m 60 -70ÿѭ ӧc tiӋ WWUQJ bҵ ng nӗi hҩ S iS ҩ t Hai VX c{QJ ӭF WK ӧ ÿѭ c bӕWUt ӟi hai Y loҥ i giӕng lӓQJ ҥ Yj ng G hҥ t MӛL F{QJ ӭF Fy WK W~L ӛL P W~L ҩ y 50g giӕ Fng hҥ t hoһ c 50ml giӕng lӓng ѬѫP ӗngWU nҩ P WURQJ Ӆ u kiӋ Q ÿL WKѭ ӡng cӫ D SKzQJ Ӌ m WKt vӟi nhiӋ W QJKL ӝÿ trung C Chi tiӃ EuQKa t vӅFKăPVyFWK ӡi kǤSKiWWUL Ӈ n sӧLYjTX ҧthӇnҩ m VzWKHR QKѭ FiFQJKLrQ ӭu ÿmWK ӵc hiӋ QWUѭ F ӟFÿk\ [12] b 3K˱˯QJ SKiSÿiQKJLiQăQJVX ̭t sinh h͕ c cͯa ṋm [13],[47] 1ăQJ ҩ t sinh VX hӑc = KhӕLOѭ ӧng nҩ P WѭѫL[NK ӕLOѭ ӧQJ QJX\rQ Ӌ u OL ... VDX /RjL Pleurotus citrinopileatus Chi: Pleurotus Hӑ: Pleurotaceae Bӝ: Agaricales Lӟp: Agaricomycetes 1JjQK ө : Basidiomycotina SK Giӟi nҩ m: Mycota (Fungi) Nҩ m VzYjQJ P citrinopileatus. .. Ho̩WWtQKNKiQJXQJWK˱F ͯa polVDFFKDULGHWDQWURQJQ ͣc ͧṋm Pleurotus citrinopileatus + HiӋ u quҧNKiQJ XQJ ӫ a Sarcoma WKѭ 180 ӣ F chuӝt cӫ D SROVDFFKD tӯnҩ m P citrinopileatus thӇhiӋ n ӣbҧ ng 1.1&ySKkQÿR... WDQ WURQJQ˱ ͣc WiFK ͳ W ṋm Pleurotus citrinopileatus B̫ng 1.2 Hi͏ u qu̫NKiQJXQJWK˱6DUFRPD ͧchu͡t cͯa SROVDFFKDULGHNK{QJWDQWURQJQ˱ ͣc WUtFKOW ͳṋm P citrinopileatus tFK Polysaccharide