Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)

Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)Nghiên cứu sàng lọc và các đặc tính sinh học của một số nấm đảm trong sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ các vùng sinh thái khác nhau (LV thạc sĩ)

B GIO DC V O TO VIN HN LM KHOA HC V CễNG NGH VIT NAM VIN SINH THI V TI NGUYấN SINH VT -*** HONG TH NHUNG NGHIấN CU SNG LC V CC C TNH SINH HC CA MT S NM M TRONG SINH TNG HP EPS, KHNG VI SINH VT, TO LACCASE T CC VNG SINH THI KHC NHAU LUN VN THC S SINH HC H Ni, 2015 B GIO DC V O TO VIN HN LM KHOA HC V CễNG NGH VIT NAM VIN SINH THI V TI NGUYấN SINH VT -*** -HONG TH NHUNG NGHIấN CU SNG LC V CC C TNH SINH HC CA MT S NM M TRONG SINH TNG HP EPS, KHNG VI SINH VT, TO LACCASE T CC VNG SINH THI KHC NHAU Chuyờn ngnh: Vi Sinh Vt Mó s: 60420103 LUN VN THC S SINH HC NGI HNG DN KHOA HC PGS TS ng Th Cm H H Ni, 2015 Lun Thc s sinh hc Hong Th Nhung- CHK17 LI CM N Trc ht, tụi xin gi li cm n chõn thnh v sõu sc ti PGS TS ng Th Cm H ó kiờn trỡ ch bo, quan tõm hng dn v dỡu dt tụi sut quỏ trỡnh hc tp, nghiờn cu v hon thnh lun vn, giỳp tụi cú thờm nhiu kin thc v kinh nghim quý bỏu nghiờn cu khoa hc Tụi xin gi li cm n n Ban Lónh o Vin Sinh thỏi v Ti nguyờn sinh vt, Ban giỏm hiu Trng i hc Thỏi Nguyờn, cựng cỏc Thy cụ giỏo ó tham gia ging dy sut khúa hc Tụi cng xin gi li cm n ti TS inh Th Thu Hng cựng th cỏn b ca phũng Cụng ngh sinh hc tỏi to mụi trng v cỏc nghiờn cu sinh ó tn tỡnh giỳp tụi ly mu v sut quỏ trỡnh thc hin lun Tụi cng xin cm n n PGS TS Thnh Th Thu Thy ó giỳp tụi b tr thờm nhng kin thc mi Cm n GS Bram Brouwer v cụng ty BioDetection Systems B.V- BDS, H Lan ó thc hin cỏc phõn tớch s dng cụng ngh DR-CALUX cú kt qu mi lun ny Cui cựng, tụi xin dnh li cm n ti chng v trai - l ch da tinh thn v nhng ngi thõn gia ỡnh l ch da vng chc, ng viờn, giỳp sut thi gian qua Mt ln na, xin chõn thnh cm n tt c nhng s giỳp quý bỏu ú ! H Ni, thỏng 12 nm 2015 Hc viờn cao hc Hong Th Nhung Trang i Lun Thc s sinh hc Hong Th Nhung- CHK17 LI CAM OAN Tụi xin cam oan õy l cụng trỡnh nghiờn cu ca tụi Cỏc s liu, kt qu nờu lun l trung thc v cha tng c cụng b bt k cụng trỡnh no khỏc Tỏc gi lun Hong Th Nhung Trang ii Lun Thc s sinh hc Hong Th Nhung- CHK17 MC LC LI CM N i LI CAM OAN ii MC LC iii BNG CH VIT TT vi DANH MC CC BNG viii DANH MC CC HèNH ix M U PHN TNG QUAN TI LIU 1.1 Tng quan v nm n v nm dc liu (MMS) .3 1.2 Lch s nghiờn cu v s dng nm 1.2.1 Tỡnh hỡnh nghiờn cu trờn th gii 1.2.2 Tỡnh hỡnh nghiờn cu Vit Nam 1.3 Polysaccharide t nm- ngun carbonhydrate cú hot tớnh sinh hc t nhiờn 1.4 Tỏch chit v xỏc nh cỏc c tớnh ca polysaccharide t nm 1.5 c im cu trỳc ca polysaccharide phõn lp t nm 11 1.6 Phng phỏp cng hng t ht nhõn (NMR) nghiờn cu cu trỳc ca polysaccharide 12 1.7 Tớnh cht vt lý ca polysaccharide 15 1.7.1 Khi lng phõn t ca polysaccharide 15 1.7.2 hũa tan ca polysaccharide t nm 15 1.8 Vai trũ ca polysaccharide t nm i vi sc khe 16 1.8.1 Tớnh khỏng u, iu tr ung th v dch ca polysaccharide 16 1.8.2 H lipid mỏu v hn ch ng huyt 20 1.8.3 Tớnh chng oxy húa ca polysaccharide t nm 21 1.8.4 Prebiotic t nm .22 1.8.5 Tớnh khỏng virus ca polysaccharide t nm 23 1.8.6 Hot tớnh khỏng khun ca polysaccharide t nm .24 1.9 Laccase v nh hng ca nú n sc khe ngi 26 Trang iii Lun Thc s sinh hc Hong Th Nhung- CHK17 1.9.1 Tng quan v laccase .26 1.9.2 ng dng ca laccase lnh vc bo v sc khe 27 1.10 Phng phỏp phõn tớch sng lc DR-Calux 28 PHN VT LIU V PHNG PHP 35 2.1 Vt liu .35 2.1.1 Vi sinh vt 35 2.1.2 Húa chõt v thit b 35 2.2 Phng phỏp nghiờn cu 36 2.2.1 Phng phỏp phõn lp cỏc chng nm 36 2.2.2 Phng phỏp phõn loi nm da vo trỡnh t xỏc nh trỡnh t ITS .36 2.2.3 Chun b ging .37 2.2.4 Phng phỏp xỏc nh sinh v hm lng exopolysaccharide 37 2.2.5 Phng phỏp xỏc nh hot tớnh enzyme laccase 37 2.2.6 ỏnh giỏ nh hng ca mt s iu kin nuụi cy n kh nng sinh trng v sinh tng hp polysaccharide v enzyme laccase 38 2.2.6.1 ỏnh giỏ anh hng ca pH .38 2.2.6.2 ỏnh giỏ nh hng ca thi gian nuụi cy .38 2.2.6.3 ỏnh giỏ nh hng ca ngun nit 39 2.2.6.4 ỏnh giỏ nh hng ca ngun carbon 39 2.2.7 Xỏc nh hm lng protein, carbonhydrate tng s ca EPS thụ 39 2.2.8 ỏnh giỏ kh nng khỏng vi sinh vt kim nh 40 2.2.9 Xỏc nh thnh phn ca polysaccharide .41 2.2.10 Xỏc nh hot tớnh ca EPS v sinh nm lờn mt s chc nng ca t bo .41 PHN KT QU V THO LUN 44 3.1 Phõn lp v phõn loi cỏc chng nm nghiờn cu .44 3.2 Kh nng sinh trng, sinh tng hp EPS v hot tớnh laccase ca cỏc chng nm .48 3.3 Kh nng khỏng vi sinh vt kim nh ca dch nuụi cy mt s chng nm 54 Trang iv Lun Thc s sinh hc Hong Th Nhung- CHK17 3.4 nh hng ca iu kin nuụi cy n s to sinh khi, sinh tng hp EPS v laccase ca chng nm Earliella sp FPT31 v Ganoderma sp FMD12 56 3.4.1 nh hng ca pH mụi trng 56 3.4.2 nh hng ca thi gian nuụi cy 59 3.4.3 nh hng ca ngun nit .61 3.4.4 nh hng ca ngun carbon 63 3.4.5 nh hng ca nng glucose 65 3.4.6 Kh nng sinh trng, sinh tng hp EPS v laccase ca FPT31 kt hp cỏc yu t mụi trng 67 3.5 Hm lng carbonhydrate v protein ca EPS thụ thu c t FPT31 v FMD12 70 3.6 Thnh phn v cu trỳc EPS ca FMD12 v FPT31 72 3.7 Hot tớnh in vitro ca EPS t chng nm FMD12 v FPT31 75 PHN KT LUN V KIN NGH 77 4.1 Kt lun 77 4.2 Kin ngh 78 TI LIU THAM KHO 79 Trang v Lun Thc s sinh hc Hong Th Nhung- CHK17 BNG CH VIT TT ABTS 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) AR Androgen Receptor CALUX Chemical Activated Luciferase Gene Expression COSY Corelated Spectrscopy CVD Cardiovascular Disease D2O Deuterium oxide dce Dried crude exopolysaccharide DMEM/F12 Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 DMSO Dimethylsulfoxide DNS 3,5-dinitrosalicylic acid tg ng tỏc gi EPS Exopolysaccharide ER Estrogen Receptor FCS Fetal calf serum GC Gas Chromatography GR Glucocorticoid Receptor HIV Human Immunodeficiency Virus HMBC Heteronuclear Multiple Bond Coherence HMQC Heteronuclear Multiple Quantum Coherence HPLC High- Performance Liquid Chromatography IC50 Concentration Inhibiting 50% Of Groth IFN Interferon IL Interleukin IPS Intracelular Polysaccharide ITS Internal transcribed spacer IZD Internal Zone Diameter Trang vi Lun Thc s sinh hc Hong Th Nhung- CHK17 NEAA Non-Essential Amino Acid MAE Microwave- Assisted Extraction MIC Minimal Inhibitory Concentration MMs Medicinal Mushrooms MRSA Methicillin-Resistant Staphylococcus aureus MRSE Methicillin-Resistant Staphylococcus epidermidis MS Mass Spectrometry MW Molecular Weight NK Natural killer NMR Nuclear Magnetic Resonance NOESY Nuclear Overhauser Effect Spectroscopy Nrf2 Nuclear factor(erythroid-derived 2)-like PBS Phosphate Buffered Saline PDA Potato Dextro Aga PDB Potato Dextro Broth PGM Potato Glucose Malt PLE Pressurized Liquid Extraction PR Progesterone Receptor PSK PolysaccaritK PSP Polysaccharopeptide ROESY Rotating Frame Overhauser Enhancement Spectroscopy SEC Size-Exclusion Chromatography SFE Supercritical Fluid Extraction TNF Tumor Necrosis Factor TSB Tryptone Soya Broth UAE Ultrasonic- Assited Extraction VREF Vancomycinresistant Enterococcus faecium Trang vii Lun Thc s sinh hc Hong Th Nhung- CHK17 DANH MC CC BNG Trang Bng 1.1 Mt s phng phỏp tỏch chit polysaccharide t nm 10 Bng 1.2 chuyn dch hoỏ hc (ppm) t c s d liu SUGABASE ca dng glucose v galactose 14 Bng 1.3 Mt s polysaccharide cú hot tớnh khỏng ung th t cỏc nm in hỡnh 19 Bng 1.4 Mt s DR- Calux thụng dng 30 Bng 2.1 Thnh phn thớ nghim ỏnh giỏ kh nng c ch s sinh trng ca vi sinh vt 36 Bng 2.2 S hỳt mu cho phõn tớch sng lc CALUX 38 Bng 3.1 tng ng gia cỏc nm thuc Ganoderma t cỏc vựng 40 Bng 3.2 Hỡnh thỏi cỏc chng nm v kh nng sinh trng, sinh tng hp polysaccharide v enzyme laccase ca chỳng 44 Bng 3.3 Kh nng c ch s sinh trng chng vi khun B cereus v M luteus ca dch nuụi cy chng FMD12 50 Bng 3.4 S sinh trng v kh nng tng hp EPS theo thi gian ca FPT31 v FMD12 55 Bng 3.5 Kh nng sinh trng, sinh tng hp EPS v laccase ca FPT31 trờn mụi trng cha ng v cao nm men nng khỏc 62 Bng 3.6 iu kin nuụi cy ca chng nm FPT31 v FMD12 vi mt s nghiờn cu quc t 64 Bng 3.7 Hm lng ng v protein 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Durieu & Mont Sapan Kumar Sharma,1 Nandini Gautam, and Narender Singh Atri Evidence-Based Complementary and Alternative Medicine Article ID 462864, pages Trang 87 ... vật nuôi trồng Xuất phát từ lý trên, đề tài Nghiên cứu sàng lọc đặc tính sinh học số nấm đảm sinh tổng hợp EPS, kháng vi sinh vật, tạo laccase từ vùng sinh thái khác nhau tiến hành Nội dung... DỤC VÀ ĐÀO TẠO VI N HÀN LÂM KHOA HỌC VÀ CÔNG NGHỆ VI T NAM VI N SINH THÁI VÀ TÀI NGUYÊN SINH VẬT -*** -HOÀNG THỊ NHUNG NGHIÊN CỨU SÀNG LỌC VÀ CÁC ĐẶC TÍNH SINH HỌC CỦA MỘT SỐ NẤM ĐẢM TRONG. .. ĐẢM TRONG SINH TỔNG HỢP EPS, KHÁNG VI SINH VẬT, TẠO LACCASE TỪ CÁC VÙNG SINH THÁI KHÁC NHAU Chuyên ngành: Vi Sinh Vật Mã số: 60420103 LUẬN VĂN THẠC SĨ SINH HỌC NGƯỜI HƯỚNG DẪN KHOA HỌC PGS TS