NGHIÊN cứu TÁCH CHIẾT BETA GLUCAN

Lun thc s khoa hc Cụng ngh Sinh hc B GIO DC V O TO TRNG I HC BCH KHOA H NI NGUYN NGC SN NGHIấN CU TCH CHIT BETA-GLUCAN T THNH T BO NM MEN BIA Chuyờn ngnh : CễNG NGH SINH HC LUN VN THC S KHOA HC CHUYấN NGNH CễNG NGH SINH HC NGI HNG DN KHOA HC: PGS TS QUN Lấ H H Ni 2013 Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc MC LC LI M U PHN I TNG QUAN V TI LIU 1.1 Tng quan v nm men bia 1.1.1 c im hỡnh thỏi ca t bo nm men 1.1.2 Cu to t bo nm men 1.2 Tng quan v betaglucan .7 1.2.1 Tớnh cht -glucan ca thnh t bo nm men S cerevisiae .7 1.2.2 Ngun nguyờn liu cha -glucan 1.2.3 Tỡnh hỡnh nghiờn cu ng dng ca -glucan 1.2.4 Cỏc ng dng ca -glucan 12 1.3 Cỏc phng phỏp thu nhn -glucan t thnh t bo nm men bia 22 1.3.1 Phng phỏp c hc phỏ v t bo nm men 22 1.3.2 Phng phỏp súng siờu õm phỏ v t bo nm men 23 1.3.3 Phng phỏp t phõn phỏ v thnh t bo nm men 23 1.3.4 Phng phỏp dựng húa cht thu nhn -glucan t thnh t bo nm men 25 1.4 Tng quan v papain (enzyme thy phõn protein nm men) 29 1.4.1 Papain t nha qu u (Carica papaya L) 29 1.4.2 Hot tớnh enzyme v nguyờn tc tỏc dng ca papain 30 PHN II VT LIU V PHNG PHP NGHIấN CU 32 2.1 Vt liu nghiờn cu 32 2.1.1 i tng nghiờn cu 32 2.1.2 Thit b s dng nghiờn cu 33 2.1.3 Húa cht s dng nghiờn cu 34 2.2 Phng phỏp nghiờn cu 34 2.2.1 Cỏc bc nghiờn cu thu nhn -glucan t t bo nm men 34 2.2.2 Phng phỏp xỏc nh m ca nm men 38 2.2.3 Phng phỏp xỏc nh hm lng Protein theo phng phỏp Kjeldahl 38 2.2.4 Phng phỏp xỏc nh hm lng carbohydrate 39 Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc 2.2.5.Phng phỏp xỏc nh hm lng -glucan (b kit Megazyme-ireland) 42 PHN III KT QU V THO LUN 47 3.1 Kho sỏt cỏc phng phỏp phỏ v t bo nm men thu nhn thnh phn ch yu ca thnh t bo nm men .47 3.1.1 T phõn nm men 47 3.1.2 Kho sỏt nh hng ca vic s dng súng siờu õm phỏ v thnh t bo nm men cỏc khong thi gian khỏc 55 3.1.3 nh hng ca vic s dng dung dch kim núng phỏ v thnh t bo nm men .56 3.1.4 Kt qu kho sỏt cỏc phng ỏn phỏ v t bo nm men khỏc thu nhn thnh t bo 57 3.2 Kho sỏt phng phỏp dựng kim tỏch protein thnh t bo nm men thu nhn phn ln -glucan .59 3.2.1 Kho sỏt nh hng ca nng NaOH ti hiu qu tỏch protein 60 3.2.2 Kho sỏt nh hng ca nhit ti hiu qu tỏch protein 61 3.2.3 Kho sỏt nh hng ca thi gian ti hiu qu tỏch protein 63 3.2.4 Kho sỏt nh hng ca t l dung mụi ti hiu qu tỏch protein 64 3.3 Nghiờn cu cỏc ch to sn phm cú hm lng -glucan cao t thnh t bo nm men bia 66 3.3.1 La chn phng ỏn tinh sch sn phm 66 3.3.2 La chn ch sy to sn phm 68 3.4 xut quy trỡnh tỏch chit v thu nhn -glucan t thnh t bo nm men bia 70 3.4.1 Thuyt minh quy trỡnh 72 PHN IV KT LUN V KIN NGH 74 4.1 Kt lun 74 4.2 Kin ngh .75 TI LIU THAM KHO 76 PH LC 82 Ph lc 82 Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc 1.1 Phng phỏp xỏc nh hm lng Protein theo phng phỏp Kjeldahl 82 Ph lc 83 2.1 Phng phỏp xỏc nh hm lng axit amin bng thuc th ninhydrin (dựng xỏc nh hot enzyme papain) 83 2.2 Phng phỏp xỏc nh hot protease ca papain 85 Ph lc 86 3.1 Kt qu thớ nghim xỏc nh hm lng -glucan 86 Ph lc 88 4.1 Hỡnh nh quỏ trỡnh tin x lý, t phõn, tỏch protein 88 Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc LI CM N hon thnh c lun ny, ngoi s c gng n lc bn thõn, tụi ó nhn c s ng h, giỳp tn tỡnh ca thy cụ giỏo, gia ỡnh v bn bố Trc ht, tụi xin by t lũng bit n chõn thnh v sõu sc ti PGS.TS Qun Lờ H - Vin Cụng Ngh Sinh Hc & Thc Phm, Trng i hc Bỏch khoa H Ni ó tn tỡnh ch bo tụi sut quỏ trỡnh hon thnh lun ny, Tụi xin chõn thnh cm n ti cỏc cụ giỏo thuc b mụn Cụng ngh Sinh hc (PGS.TS Lờ Thanh H, PGS TS Nguyn Lan Hng), PGS.TS Khut Hu Thanh Trng i hc Bỏch khoa H Ni ó ging dy v giỳp tụi sut quỏ trỡnh hc v thc hin lun Xin c cm n n ton th cỏn b, cụng nhõn viờn Cụng ty c phn Bia H Ni- Hi Dng ó giỳp v to iu kin cho tụi lm nghiờn cu sut quỏ trỡnh hc v lm lun ng thi tụi xin chõn thnh cm n gia ỡnh tụi v cỏc thy cụ cựng cỏc bn hc viờn, sinh viờn lm vic ti phũng thớ nghim thuc Vin Cụng ngh Sinh hc & Thc Phm, Trng i hc Bỏch khoa H Ni ó to iu kin, quan tõm, ng viờn v gúp ý cho tụi sut quỏ trỡnh hc tp, nghiờn cu v hon thnh lun H Ni, ngy 25 thỏng nm 2013 Hc viờn Nguyn Ngc Sn Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc LI CAM OAN Tụi l Nguyn Ngc Sn xin cam oan ni dung quyn lun ny vi ti Nghiờn cu tỏch chit Beta-Glucan t thnh t bo nm men bia' l cụng trỡnh nghiờn cu v sỏng to chớnh tụi thc hin di s hng dn ca PGS.TS Qun Lờ H - B mụn Cụng ngh Sinh hc Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc DANH MC CC Kí HIU, CC CH VIT TT STT Ký hiu Tờn y Ghi chỳ BYC Brewers yeast whole cells Nm men bia sau tin x lý MOS Mannan-Oligo-Saccharide v/phỳt Vũng/phỳt PA Phng ỏn SP Sn phm LDL Low Density Lipoprotein Lipoprotein ty trng thp DANH MC BNG, BIU STT Tờn bng biu Trang Bng 1.1 Cu trỳc ca -glucan c tỏch t mt s ngun khỏc Bng 1.2 Cỏc -glucan cú hot tớnh sinh hc thng c s dng Bng 1.3 Mt s ng dng in hỡnh ca -glucan thc phm 11 19 Bng 2.1 Hot lc ca enzyme papapain 33 Bng 2.2 B trớ thớ nghim s dng papain thy phõn nm men 37 Bng 2.3 Cỏch pha nng glucose xõy dng ng chun 40 Bng 2.4 Giỏ tr OD ca dung dch glucose nng khỏc 41 10 Bng 3.1 nh hng ca thi gian n kt qu thy phõn ca nm men Bng 3.2 nh hng ca nhit n kt qu thy phõn ca nm men Bng 3.3 nh hng ca pH n kt qu thy phõn ca nm men Nguyn Ngc Sn 48 49 51 i hc Bỏch khoa H Ni Lun thc s khoa hc 11 12 13 14 15 16 17 18 19 20 21 22 Cụng ngh Sinh hc Bng 3.4.nh hng ca hm lng papain n hiu sut thy phõn Bng 3.5.nh hng ca vic s dng enzyme thy phõn sau nm men ó c t phõn Bng 3.6 nh hng ca quỏ trỡnh x lý bng song siờu õm cỏc thi gian khỏc Bng 3.7 Kt qu ca phỏ v t s b thnh t bo nm men bng NaOH Bng 3.8.Tng hp kt qu kho sỏt x lý t bo nm men bia Bng 3.9 kho sỏt nh hng nng NaOH n hm lng protein, carbohydrate v hm lng cht rn Bng 3.10 Kho sỏt nh hng ca nhit n n hm lng protein, carbohydrate v hm lng cht rn Bng 3.11 Kho sỏt nh hng ca thi gian n hm lng protein, carbohydrate v hm lng cht rn Bng 3.12 Kho sỏt nh hng ca ty l dung mụi n hm lng protein, carbohydrate v hm lng cht rn Bng 3.13 Tng hp nghiờn cu cỏc ch x lý nm men nhng iu kin khỏc to sn phm cú cha -glucan Bng 3.14 Kho sỏt cỏc ch sy Nguyn Ngc Sn 54 55 57 58 60 62 63 65 67 69 Bng 3.15 Thnh phn sn phm bng phng phỏp sy khỏc 53 70 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc DANH MC CC HèNH V, TH STT Tờn hỡnh v, th Trang Hỡnh 1.1 Cu to t bo nm men Hỡnh 1.2 Cu to thnh t bo nm men Hỡnh 1.3 Cu trỳc -glucan ca thnh t bo nm men 7 Hỡnh 1.4 Dectin-1 trung gian cho hiu qu sinh hc ca glucan Hỡnh 1.5 S phỏ v thnh t bo nm men bng phng phỏp c hc Hỡnh 1.6 S thu nhn -glucan t t bo nm men S cerevisiae Hỡnh 1.7 S thu nhn cỏc phõn on -glucan t t bo nm men S cerevisiae Hỡnh 2.1 Bó thi nm men bia 10 23 26 28 32 Hỡnh 2.2 Cỏc bc c bn thu nhn -glucan t thnh t bo nm men bia 35 10 Hỡnh 2.3 Phng trỡnh to mu gia glucose v Anthrone 40 11 Hỡnh 3.1 Sn phm sau sy thng 69 12 Hỡnh 3.2 Sn phm sau sy ụng khụ 69 13 Hỡnh 3.3 Sn phm sau sy phun 69 14 15 Hỡnh 3.4 Quy trỡnh cụng ngh thu nhn sn phm -glucan t thnh t bo nm men bia th 2.2 Phng trỡnh ng chun glucose bc súng =630nm Nguyn Ngc Sn 71 41 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc LI M U Trong nhng nm gn õy Vit Nam cng nh trờn th gii nhu cu phỏt trin, s lng cỏc nh mỏy ch bin ru, bia, ngy cng mt gia tng em li giỏ tr to ln v m t kinh t cho t nc Tuy nhiờn c ng vi s phỏt trin ny, mt lng bó men thi khng l thi ngoi mụi trng Trung bỡnh c sn xut 1000 lớt bia thỡ x 10 kg men thi vi m trung bỡnh khong 85% Lng nm men ny ch yu d ng ch bin thc n ng vt hay c d ng nh l mt ngun nguyờn liu r tin lm dch chit nm men t phõn, phn cũn li thi ngoi mụi trng gõy ụ nhim n mụi trng sng xung quanh [20,56] Trong t bo nm men, thnh t bo chim khong 10-20% trng lng khụ ca t bo, ú glucan chim khong 50-60% lng cht khụ ca thnh t bo nm men Chớnh vỡ vy, vic sn xut cỏc sn phm ph, cú giỏ tr kinh t nh -glucan cú th mang li li nhun cao cho cỏc nh mỏy xớ nghip ng thi cũn cú tỏc ng lm gim thiu tỏc ng v m t mụi trng t nhng nh mỏy cụng nghip ny Nhiu nc trờn th gii nh Hn Quc, M, Nht bn cng ó tin hnh nghiờn cu tỏch chit -glucan sch t thnh t bo nm men s dng nh yu t kớch thớch h dch v tỏc ng tớch cc n h thng bo v vt ch, tng tớnh khỏng ca vt ch n hu ht cỏc loi bnh nhim khun, nm, virut c ng nhiu loi sinh vt ký sinh khỏc Ngoi ra, cỏc nh khoa hc Nht bn cng ó s dng glucan nh l mt yu t dch hiu qu iu tr ung th [27] Bờn cnh ú, glucan cũn l cht chng oxy hoỏ, tỏi to da, lm m np nhn lờn c d ng lm cht b sung sn xut m phm [39] Vit Nam, õy l mt hng nghiờn cu cũn mi ú chỳng tụi bc u nghiờn phng phỏp tỏch chit -glucan t thnh t bo nm men bia S cerevisiae t tn thu ngun bó thi nm men bia nhm s dng ngnh cụng nghip thc phm, dc phm v m phm Nguyn Ngc Sn i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc cũn cú kh nng tn thu c ngun axit amin t quỏ trỡnh thy phõn nm men lm nguyờn liu ph gia cú giỏ tr cao v dinh dng cú th ng dng cụng ngh thc phm, sinh hc 4.2 Kin ngh Trong phm vi ti chỳng tụi ch nghiờn cu khuụn kh nm men bia thi ca Cụng ty c phn Bia H Ni Hi Dng v nghiờn cu mt s cỏc yu t tỏch -glucan t thnh t bo nm men - ngh c tip tc 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Klis*, Andre Boorsma and Piet W J De Groot Swammerdam (2006), Cell wall construction in Saccharomyces cerevisiae, Institute for Life Sciences, University of Amsterdam, BioCentrum Amsterdam, The Netherlands Yeast 2006; 23: 185202., Nguyn Ngc Sn 80 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc 52 Aguilar-Uscanga, B Francois JM (2003) A study of the yeast cell wall composition and structure in response to growth conditions and mode of cultivation, Lett Appl Microbiol 37: 268274 53 Klis FM, Mol P, Hellingwerf K, Brul S (2002) Dynamics of cell wall structure in Saccharomyces cerevisiae, FEMS Microbiol Rev 26: 239-256 54 Lipke PN, Ovalle R (1998), Cell wall structure and new challenges, J Bacteriol architecture in yeast new 180:37353740 55 Vesna ZECHNER-KRPAN (2009), Potential Application of Yeast - Glucans in Food Industry, review article 56 M Suphantharika *, P Khunrae, P Thanardkit, C Verduyn (2002) Preparation of spent brewers yeast b-glucans with a potential application as animmunostimulant for black tiger shrimp, Penaeus monodon, Department of Biotechnology, Faculty of Science, Mahidol University, Rama Road, Bangkok 10400, Thailan 57 Celine Laroche and Philippe Michaud (2007) New Developments and Prospective Applications for (1,3) Glucans,Recent Patents on Biotechnology 2007, 1, 59-73 58 Morris GJ, Winters L, Coulson GE, Clarke KJ (1986) Effect of osmotic stres on the ultrastructure and viability of the yeast Saccharomyces cerevisiae J Gen Microbiol 132: 20232034 59 Wolfgang Kunze (2009), Technology brewing and malting, international edition C NGUN INTERNET 60 http://vi.wikipedia.org/wiki/Beta-glucan 61 http://en.wikipedia.org/wiki/Sonication 62 http://biochemie.web.med.uni-muenchen.de/Yeast_Biol Nguyn Ngc Sn 81 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc PH LC Ph lc 1.1 Phng phỏp xỏc nh hm lng Protein theo phng phỏp Kjeldahl * Nguyờn tc: Khi t núng nguyờn liu em phõn tớch vi H2SO4 m c, cỏc hp cht hu c b oxy hoỏ Cacbon v hydro to thnh CO2 v H2O cũn nit sau c gii phúng di dng NH3 kt hp vi H2SO4 to thnh (NH4)2SO4 tan dung dch [11] ui amoniac dung dch bng NaOH, ng thi ct v thu nú bng lng d axit boric H3PO3 (NH4)2SO4 + 2NaOH 2NH4OH + Na2SO4 2NH4OH + H3BO3 (NH4)2B4O7 + 7H2O nh phõn lng tetraborat amụn to thnh bng dung dch H2SO4 chun, qua ú tớnh c lng nit mu vt (NH4)2B4O7 + H2SO4 + 5H2O (NH4)SO4 + H3BO3 * Cỏch thc thc hin: - Cõn 0,5g mu ó ct nh chuyn vo bỡnh Kjeldahl - Cho tip vo bỡnh Kjeldahl 10ml H2SO4 m c (d=1,84) tng nhanh quỏ trỡnh vụ c hoỏ (t chỏy) cn phi cho thờm hn hp xỳc tỏc CuSO4 v K2SO4 ( ty l 1:3) Sau ú un cho n dung dch hon ton mt mu v ngui - Chuyn ton b dung dch sau ó vụ c hoỏ xong bỡnh Kjeldahl vo bỡnh nh mc 100 ml thờm nc ct n vch mc, lc u - Ly 10 ml dung dch thớ nghim sau ó vụ c húa c nh mc n 100ml v 10ml dung dch NaOH 40% cho vo bu ct Quỏ trỡnh ct kt thỳc sau 15 phỳt, nh phõn lng tetraborat amụn to thnh bng dung dch H2SO4 0,01N Ct mt mu trng kim chng vi cỏc thuc th v thao tỏc nh trờn nhng thay 10 ml dung dch ó vụ c hoỏ bng 10 ml nc ct Kt qu c tớnh theo cụng thc: Nguyn Ngc Sn 82 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc Trong ú : X: Hm lng protein (%) 6,25: H s chuyn i sang hm lng protein a: S ml H2SO4 0,01N d ng chun mu thớ nghim b: S ml H2SO4 0,01N d ng chun mu kim chng 0,14: mg nit ng vi 1ml H2SO4 0,01N V: Dung tớch bỡnh nh mc V1: ml dung dch thớ nghim hỳt t bỡnh nh mc vo bu ct m: Lng mu em ct m (mg) Ph lc 2.1 Phng phỏp xỏc nh hm lng axit amin bng thuc th ninhydrin (dựng xỏc nh hot enzyme papain) Bn cht ca phng phỏp l dung ninhydrin phn ng vi axit amin (dng -aminoaxit) to phc cht mu xanh tớm Tin hnh o mu trc tip cú s dng pyridine l cht n nh mu[10] Axit glutamic l mt -amin tham gia phn ng to mu vi ninhydrin cho mu xanh v c o bc súng 570nm Cú th túm lc quy trỡnh ng chun axit glutamic theo s : Pha dung dch 0,1% (0,1g/100ml) Dung dch aa phõn tớch 100 ml Ly 10 ml nh mc n 100 ml Pha cỏc nng t 0,01g/l-0,1g/l Pha loóng Ly 1ml + 1ml ninhydrin 2% + 1ml pyridine 20% Xỏc nh mt quang bc súng 570 nm un núng 750C 10 phỳt Hỡnh 2.1.1 Cỏc bc tin hnh xỏc nh hm lng axit amin v xõy dng ng chun axit glutamic Nguyn Ngc Sn 83 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc Dung dch sau c pha theo nng 0,01g/l n 0,1g/l c tin hnh phn ng vi ninhydrin Ly 1ml dch cng vi 1ml pyridine 20% v 1ml ninhydrin 2%, un núng nhit 750C 10 phỳt o mt quang bc súng 570 nm Dng ng chun v cú phng trỡnh chun ng vi R2 C th nh sau Giỏ tr mt quang xõy dng ng chun axit glutamic Bng 2.1.1.Giỏ tr OD ca dung dch axit glutamic cỏc nng khỏc Nng Kt qu o OD(570nm) 0,001 0,041 0,002 0,123 0,003 0,232 0,004 0,335 0,05 0,402 0,06 0,497 th 2.1.1.Phng trỡnh ng chun axit glutamic Phng trỡnh ng chun axit glutamic cú dng y=9.2x-0.050 Nguyn Ngc Sn 84 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc 2.2 Phng phỏp xỏc nh hot protease ca papain Mt n v hot lc ca enzyme protease c nh ngha l gram axit amin c to sau phn ng thy phõn bng enzyme vi c cht l gelatin u kin nhit 500C 30 phỳt Hm lng axit amin c xỏc nh bng phn ng vi ninhydrin v tớnh theo axit glutamic Phng phỏp xỏc nh hot enzyme protease cú th túm lc bng s sau [10] Pha dung dch enzyme papain (0,1g/100ml) Chun b dung dch gelatin nng 2% Ly 1ml dung dch E + 4ml gelatin n nhit 500C 30 phỳt Ly 1ml dch, pha loóng 25 ln Ly 1ml dch + 1ml ninhydrin 20% + 1ml pyridine 20% un núng nhit 750C 10 phỳt Xỏc nh mt quang bc súng 570 nm hỡ Hỡnh 2.2.1 Cỏc bc tin hnh xỏc nh hot lc ca protease Kt qu o mt quang thu c, s dng phng trỡnh ng chun glutamic tớnh hot lc ca enzyme ph mc [2.2.5] Hot ca enzyme c tớnh theo cụng thc: Trong ú: C: g/l tớnh t phng trỡnh ng chun n: h s pha loóng 5: th tớch ng 5ml 10: thi gian phn ng 1000/147 h s quy i theo axit glutamic Nguyn Ngc Sn 85 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc 2.3 Bng xỏc nh hm hot lc enzyme papain Giỏ tr mt H s pha Lng aa sau Hot lc enzyme quang (570nm) loóng thy phõn (g) (U/g) 0,55 250 16,7935 5545 0,59 250 17,1739 5915 0,565 250 17,1195 5684 Ph lc 3.1 Kt qu thớ nghim xỏc nh hm lng -glucan Bng 3.1 phõn tớch hm lng - glucan Giỏ tr Mu Khi lng Hm lng mt quang m mu phõn (510nm) % tớch (mg) BYC 0,0480,004 100 4,04 SP1 0,210 0,03 100 18,650,53 SP4 0,2800,03 100 23,640,64 SP5 0,2700,04 100 22,350,45 SP6 0,2800,02 100 23,500,51 SP7 0,2810,008 100 23,670,36 SP8 0,2820,008 100 23,870,40 SP9 0,2820,008 100 23,870,53 Nguyn Ngc Sn 86 -glucan 0,68 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc Bng 3.2 phõn tớch hm lng glucan tng s Giỏ tr Mu Khi lng Hm lng mt quang m mu phõn tớch Glucan (510nm) % (mg) tng s BYC 0,0380,003 100 17,530,667 SP1 0,480 0,05 100 42,380,53 SP4 0,9400,03 100 82,660,64 SP5 0,9800,04 100 81,870,45 SP6 1,010 0,03 100 83,490,51 SP7 1,0100,04 100 83,470,40 SP8 1,0200,02 100 85,230,40 SP9 1,0200,03 100 85,240,52 Nguyn Ngc Sn 87 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc Ph lc Bng 4.1 S nh hng ca thi gian ngõm dung dch NaOH cỏc khong thi gian khỏc n hm lng carbohydrate, protein, cht rn Thi gian Hm lng Hm lng (Phỳt) carbohydrate protein (g) ( g) (Nì6,25) 30 2,950,04 0,2090,002 3,620,065 60 2,230,29 0,0880,004 2,460,046 90 1,950,23 0,0820,015 2,170,042 120 1,890,19 0,0780,003 2,100,051 SP1 4,850,053 5,020,016 10 Hm lng cht rn (g) 4.1 Hỡnh nh quỏ trỡnh tin x lý, t phõn, tỏch protein Nm men bia thi Nguyn Ngc Sn 88 i hc Bỏch khoa H Ni Lun thc s khoa hc Cụng ngh Sinh hc Nm men sau tin x lý Nm men sau t phõn Sn phm sau x lý bng NaOH Nguyn Ngc Sn 89 i hc Bỏch khoa H Ni ... PHƯƠNG PHÁP NGHIÊN CỨU 32 2.1 Vật liệu nghiên cứu 32 2.1.1 Đối tượng nghiên cứu 32 2.1.2 Thiết bị sử dụng nghiên cứu 33 2.1.3 Hóa chất sử dụng nghiên cứu ... Nguyễn Ngọc Sơn xin cam đoan nội dung luận văn với đề tài Nghiên cứu tách chiết Beta-Glucan từ thành tế bào nấm men bia’' công trình nghiên cứu sáng tạo thực hướng dẫn PGS.TS Quản Lê Hà - Bộ môn... 1.2.3.2 Tình hình nghiên cứu β-glucan Việt Nam Hiện nguồn β-glucan chủ yếu nhập từ nước sử dụng nhiều lĩnh vực Việc nghiên cứu β-glucan Việt Nam mẻ, đề tài cấp nhà nước năm 2005 Nghiên cứu xây dựng