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Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)Nghiên cứu sử dụng kết hợp enzyme trong chiết tách và làm giàu một số sản phẩm nguồn gốc thiên nhiên (LA tiến sĩ)
B GIO DC V O TO VIN HN LM KHOA HC V CễNG NGH VIT NAM HC VIN KHOA HC V CễNG NGH HONG TH BCH Nghiên cứu sử dụng kết hợp enzyme chiết tách làm giàu số sản phẩm nguồn gốc thiên nhiên LUN N TIN S HO HC H NI - 2017 B GIO DC V O TO VIN HN LM KHOA HC V CễNG NGH VIT NAM HC VIN KHOA HC V CễNG NGH HONG TH BCH Nghiên cứu sử dụng kết hợp enzyme chiết tách làm giàu số sản phẩm nguồn gốc thiên nhiên Chuyờn ngnh: Húa hc cỏc hp cht t nhiờn Mó s: 62.44.01.17 LUN N TIN S HO HC Ngi hng dn khoa hc: PGS.TS Lờ Mai Hng PGS.TS Nguyn Quyt Chin H NI - 2017 i LI CAM OAN Tụi xin cam oan õy l cụng trỡnh nghiờn cu ca bn thõn, c hỡnh thnh v phỏt trin t nhng quan im ca cỏ nhõn tụi, di s hng dn ca PGS.TS Lờ Mai Hng v PGS.TS Nguyn Quyt Chin, cú tham kho thờm cỏc ti liu ỏng tin cy, cú ngun gc rừ rng Cỏc s liu, kt qu lun ỏn l hon ton trung thc v cha c cụng b bt k cụng trỡnh no khỏc Tụi xin hon ton chu trỏch nhim v ni dung lun ỏn ny Tỏc gi lun ỏn Hong Th Bớch ii LI CM N u tiờn, tụi xin by t lũng bit n chõn thnh v sõu sc nht ti PGS TS Lờ Mai Hng v PGS TS Nguyn Quyt Chin, nhng ngi thy bng c tõm huyt ca mỡnh ó hng dn tụi v khoa hc, gi m cho tụi cỏc ý tng nghiờn cu v chia s nhiu ca cuc sng sut thi gian tụi nghiờn cu lun ỏn Tụi xin gi li cm n ti GS TS Phm Quc Long - Vin trng Vin Húa hc cỏc Hp cht thiờn nhiờn, cựng ban lónh o Vin, b phn o to ca Vin Húa hc cỏc Hp cht thiờn nhiờn ó to iu kin rt nhiu cho tụi hon thnh lun ỏn ca mỡnh Tụi xin gi li cm n ti cỏc thy, cỏc anh ch ph trỏch ca Hc Vin Khoa hc Cụng ngh, ó to mi iu kin cho tụi hon thnh lun ỏn ca mỡnh Tụi xin gi li cm n ti cỏc anh ch em ng nghip Trung tõm Nghiờn cu v Phỏt trin cỏc SPTN ó to iu kin giỳp tụi quỏ trỡnh hon thnh cụng trỡnh nghiờn cu ny Tụi cm n ti Nghiờn cu ng dng cụng ngh sinh hc to ch phm acid bộo a ni ụi (n3 - PUFA) t nguyờn liu t nhiờn b sung vo thc n ng nuụi mt s i tng cỏ bin ch lc v Nghiờn cu thnh phn húa hc, iu tra v ỏnh giỏ cht lng tinh du trm hin ang c sn xut Vit Nam, mó s VAST 04; ó ti tr kinh phớ Tụi xin gi li tri õn ca mỡnh ti gia ỡnh, bn bố, nhng ngi thõn luụn ng viờn tụi cú ng lc cụng vic v hon thnh cụng trỡnh nghiờn cu khoa hc ny Xin chõn thnh cm n! Tỏc gi Hong Th Bớch iii MC LC Trang Li cam oan i Li cm n ii Mc lc iii Danh mc cỏc ký hiu, cỏc ch vit tt vii Danh mc bng ix Danh mc hỡnh v .x M U CHNG TNG QUAN TI LIU 1.1 Tng quan v ng dng cụng ngh enzyme - gii phỏp Húa hc xanh chit xut v lm giu cỏc sn phm cú ngun gc thiờn nhiờn 1.1.1 Húa hc xanh chit xut v lm giu cỏc sn phm cú ngun gc thiờn nhiờn 1.1.2 C s ng dng enzyme h tr chit xut cỏc sn phm cú ngun gc thiờn nhiờn 1.1.3 Mt s yu t nh hng n quỏ trỡnh thy phõn bi enzyme 1.1.4 Li ớch v khú khn ng dng cụng ngh EAE 1.2 Cỏc enzyme s dng cụng ngh enzyme h tr chng ct (EAD) v enzyme h tr chit xut (EAE) 10 1.2.1 Cỏc enzyme phõn gii cu trỳc thnh t bo nguyờn liu lignocellulose 10 1.2.2 Enzyme protease phõn gii cu trỳc nguyờn liu giu protein 16 1.2.3 Enzyme lipase thy phõn lipid thnh axit bộo t 19 1.3 Tỡnh hỡnh nghiờn cu ng dng cụng ngh enzyme chit xut v lm giu cỏc hp cht thiờn nhiờn v ngoi nc 22 1.3.1 Cỏc nghiờn cu trờn th gii 22 1.3.2 Cỏc nghiờn cu nc 26 1.4 Tng quan v qu Cinnamomum cassia 27 iv 1.4.1 Gii thiu v Qu C.cassia 27 1.4.2 Nhng nghiờn cu v thnh phn húa hc C cassia 29 1.4.3 Nhng nghiờn cu v hot tớnh sinh hc C cassia 31 1.4.4 Cỏc phng phỏp chit xut tinh du qu 33 1.5 Tng quan v cỏ ng 35 1.5.1 Sn lng v giỏ tr dinh dng ca ph phm cỏ ng 35 1.5.2 Du cỏ v cỏc axit bộo khụng no a ni ụi n-3 PUFA 37 1.5.3 Cỏc phng phỏp chit xut lipid v lm giu cỏc axit bộo khụng no 39 CHNG NGUYấN VT LIU V PHNG PHP NGHIấN CU .41 2.1 Nguyờn liu 42 2.1.1 Mu nguyờn liu 42 2.1.2 Enzyme s dng nghiờn cu 43 2.1.3 B chng vi sinh vt kim nh 44 2.1.4 Dũng t bo 45 2.2 Húa cht, thit b 45 2.2.1 Húa cht 45 2.2.2 Thit b 46 2.3 Phng phỏp nghiờn cu 46 2.3.1 Cỏc phng phỏp phõn tớch húa sinh 46 2.3.2 Cỏc phng phỏp phõn tớch thnh phn húa hc 46 2.3.3 Cỏc phng phỏp xỏc nh hot enzyme 47 2.3.4 Cỏc phng phỏp thy phõn 49 2.3.5 Cỏc phng phỏp xỏc nh sn phm thy phõn 50 2.3.6 Phng phỏp lm giu cỏc axit bộo bng kt tinh ure 52 2.3.7 Cỏc phng phỏp nghiờn cu iu kin ti u 52 2.3.8 Cỏc phng phỏp xỏc nh hot tớnh sinh hc 53 CHNG THC NGHIM .54 3.1 Nghiờn cu ng dng enzyme kt hp chng ct tinh du t cỏc b phn khỏc ca cõy qu Cinnamomum cassia 54 v 3.1.1 Nghiờn cu phõn tớch thnh phn cht nn v tinh du ca mu cnh lỏ v mu v qu C cassia 55 3.1.2 Nghiờn cu tỏc ng ca vic x lý nguyờn liu vi enzyme lờn quỏ trỡnh chng ct tinh du 55 3.1.3 Nghiờn cu tỡm iu kin ti u cho quỏ trỡnh ng dng h enzyme kt hp Laccase-Htec2 58 3.2 Nghiờn cu thm dũ ng dng h enzyme kt hp Laccase-Htec2 chng ct tinh du trm hng t g cõy giú bu Aquilaria crassna 60 3.2.1 Nghiờn cu phõn tớch thnh phn cht nn ca bt g giú bu A crassna 60 3.2.2 Nghiờn cu thm dũ tỏc ng ca vic x lý enzyme Laccase- Htec2 lờn quỏ trỡnh chng ct tinh du t bt g giú bu A crassna 60 3.3 Nghiờn cu ng dng enzyme kt hp chit xut lipid v lm giu cỏc axit bộo n-3 PUFA t u cỏ ng võy vng Thunnus albarcares 61 3.3.1 Nghiờn cu thnh phn cht nn v lipid tng ca mt s loi u cỏ ng ca Vit Nam 62 3.3.2 Nghiờn cu ng dng enzyme protease chit xut lipid t u cỏ ng võy vng T albarcares 62 3.3.3 Nghiờn cu ng dng enzyme lipase kt hp ure quỏ trỡnh lm giu cỏc axit bộo n-3 PUFA ca du u cỏ ng võy vng T albacares 64 CHNG KấT QU V THO LUN 66 4.1 Kt qu nghiờn cu ng dng enzyme kt hp chng ct tinh du t cỏc b phn khỏc ca cõy qu Cinnamomum cassia 66 4.1.1 Kt qu phõn tớch cỏc cht nn v tinh du ca cnh lỏ v v qu C cassia 66 4.1.2 Tỏc ng ca vic x lý nguyờn liu vi enzyme lờn quỏ trỡnh chng ct tinh du t cnh lỏ qu C cassia 71 4.1.3 Tỏc ng ca vic x lý nguyờn liu vi enzyme lờn quỏ trỡnh chng ct tinh du t v qu C cassia 85 vi 4.1.4 Kt qu tỡm iu kin ti u cho quỏ trỡnh ng dng h enzyme kt hp Laccase-Htec2 chng ct tinh du t cnh lỏ v v qu 95 4.2 Kt qu thm dũ ng dng h enzyme kt hp Laccase-Htec2 chng ct tinh du trm hng t g cõy giú bu Aquilaria crassna 106 4.2.1 Kt qu phõn tớch thnh phn cht nn ca bt g giú bu A crassna 106 4.2.2 Kt qu thm dũ tỏc ng h enzyme Laccase - Htec2 lờn hm lng v thnh phn tinh du trm hng t g cõy giú bu A crassna 107 4.3 Kt qu nghiờn cu ng dng enzyme kt hp chit xut lipid v lm giu cỏc axit bộo n-3 PUFA t u cỏ ng võy vng Thunnus albarcares 111 4.3.1 Kt qu nghiờn cu cỏc cht nn v thnh phn axit bộo ca mt s loi u cỏ ng ca Vit Nam 111 4.3.2 Kt qu nghiờn cu ng dng enzyme chit xut lipid t u cỏ ng võy vng T albarcares 115 4.3.3 Kt qu nghiờn cu ng dng enzyme lipase CRL quỏ trỡnh lm giu cỏc axit bộo n-3 PUFA ca du u cỏ ng võy vng T albacares bng phng phỏp ure 118 4.3.4 xut qui trỡnh cụng ngh ng dng enzyme kt hp chit xut lipid v lm giu n-3 PUFA t u cỏ ng võy vng T albacares 126 KT LUN 130 KIN NGH 132 DANH MC CC CễNG TRèNH CễNG B CA TC GI LIấN QUAN N LUN N 133 DANH MC TI LIU THAM KHO 134 PH LC vii DANH MC CC Kí HIU, CC CH VIT TT Ký hiu Ting Anh Ting Vit ALA : Alpha linolenic acid Axit Alpha linolenic CMC : Carboxymethylcellulose Carboxymethylcellulose DHA : Docosahexaenoic acid Axit Docosahexaenoic DMSO : Dimethyl Sulphoxide Dimethyl Sulphoxide DNS : 3,5-dinitrosalicylic acid 3,5-dinitrosalicylic acid DPA : Docosapentaenoic acid Axit Docosapentaenoic DPPH : Diphenylpicrylhydrazyl Diphenylpicrylhydrazyl EAD : Enzyme assisted distillation Chng ct cú enzyme h tr EAE : Enzyme assisted extraction Chit xut cú enzyme h tr EAMD : Enzyme assisted microway extraction Chit xut cú enzyme kt hp vi súng EDTA : Ethylene Diamine Triacetic Acid Ethylene Diamine Triacetic Acid EPA : Eicosapentaenoic acid Axit Eicosapentaenoic FAO : Food and Agriculture Organization of T chc Lng nụng Th gii the United Nations FPH : Fish protein hydrolysate Protein thy phõn t cỏ GC : Gas chromatography Sc ký khớ GC-MS : Gas chromatography mass spectrometry Sc ký kt hp ph HD : Hydrodistillation Chng ct lụi cun hi nc Hep-G2 : Human hepatocellular carcinoma Dũng t bo ung th gan HPLC : High Performanc/e liquid Sc ký lng hiu nng cao chromatography Htec2 : Cellic HTec2 Enzyme Enzym Cellic HTec2 IC50 : Inhibitory concentration at 50% Nng c ch 50% KPH : Khụng phỏt hin Laccase : Laccase enzyme Enzym laccase viii LD100 Liu lng gõy cht cho 100% s : Lethal Dose at 100% ng vt th nghim LU : Human lung adenocarcinoma Dũng t bo ung th phi 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CễNG NGH VIT NAM HC VIN KHOA HC V CễNG NGH HONG TH BCH Nghiên cứu sử dụng kết hợp enzyme chiết tách làm giàu số sản phẩm nguồn gốc thiên nhiên Chuyờn ngnh: Húa hc cỏc hp cht t nhiờn Mó s: 62.44.01.17... Diphenylpicrylhydrazyl EAD : Enzyme assisted distillation Chng ct cú enzyme h tr EAE : Enzyme assisted extraction Chit xut cú enzyme h tr EAMD : Enzyme assisted microway extraction Chit xut cú enzyme kt hp vi... trỡnh thy phõn bi enzyme 1.1.4 Li ớch v khú khn ng dng cụng ngh EAE 1.2 Cỏc enzyme s dng cụng ngh enzyme h tr chng ct (EAD) v enzyme h tr chit xut (EAE) 10 1.2.1 Cỏc enzyme phõn gii