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(Extraction of biological activities from Ganoderma lucidum
and examination of bioactivities)
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Trang 3
7rQWLӃng Anh: Extraction of biological activities from Ganoderma lucidum
and examination of bioactivities
II NHIӊM VӨ 9¬1ӜI DUNG:
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Trang 4LӠI CҦM Ѫ1
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Trang 57Ï07ҲT LUҰ19Ă1
Trong luұQ YăQ Qj\ WULWHUSHQRLG ÿѭӧF WUtFK O\ Wӯ nҩm linh chi ViӋt Nam bҵng SKѭѫQJ SKiS HQ]\PH NӃt hӧS VLrX kP +jP OѭӧQJ WULWHUSHQRLG ÿѭӧF [iF ÿӏnh bҵng SKѭѫQJSKiSSKiSTXDQJSKә hҩSWKXSKkQWӱ (UV±Vis)
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khҧ QăQJ NKiQJ NKXҭn cӫa triterSHQRLG ÿѭӧc thӱ nghiӋP WUrQ Escherichia coli,
Staphylococcus aureus, Pseudomonas aeruginosa Yj Salmonella enterica bҵng
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Trang 6ABSTRACT
In this thesis, triterpenoid was extracted from Vietnamese Ganoderma lucidum (G
lucidum) by the ultrasonic-assisted enzyme extraction (UAEE) method The
triterpenoid content was determined by ultraviolet±visible spectroscopy (UV±Vis)
Chemical composition and heavy metal content in G lucidum have been investigated, showing that G lucidum has high fiber, carbohydrate and lipid content
In addition, the content of heavy metals such as As, Cd, Hg and Pb was within the allowable limit
The influence of each extraction condition such as enzyme concentration, etanol:material ratio, temperature, time and power on triterpenoid content was investigated by the method of alternating each variable to find a suitable range of variation Then, conducting a simultaneous survey by experimental planning using the Plackett±Burman and Box±Behnken designs The data were processed using Design Expert 11.0 software to find extraction conditions for the highest triterpenoid content
At the same time, the characteristic functional groups of the extract, the morphology of the surface structure of the fungus and the high composition of the extract were investigated by Fourier transform infrared spectroscopy (Fourier transform infrared spectroscopy ± FTIR), scanning electron microscopy (scan electron microscopy ± SEM), and gas chromatography ± mass spectrometry (GC±MS)
The extract at optimal conditions was tested for biological activity through antitumor activity on three human cancer cell lines (Hep-G2 and MCF-7) and the
antibacterial ability of triterpenoids was tested on Escherichia coli, Staphylococcus
aureus, Pseudomonas aeruginosa, and Salmonella enterica by antibacterial ring
diameter measurement
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Trang 81.1.1 Giͣi thi͏u chung 2
Trang 91.5.4 ĈyQJJySFͯa lu̵QYăQ 28
THӴC NGHIӊM 29
&+ѬѪ1*+yDFKҩt, dөng cө, thiӃt bӏ YjÿӏDÿLӇm thӵc hiӋn 29
+RiFK̭t 29
2.1.2 Thi͇t b͓ 29
Ĉ͓DÿL͋m thc hi͏n lu̵QYăQ 30
7KtQJKLӋm 30
2.2.1 Kh̫RViWWKjQKSK̯QKRiK͕c trong ṋm linh chi 30
2.2.2 Kh̫RViWV ̫QKK˱ͧng cͯDFiF\͇u t͙ ÿ͇QKjPO˱ͫng triterpenoid 33
3KkQWtFKÿ̿F WtQKKyD OêWKjQKSK̯n cͯDFDRWUtFKYj F̭X WU~FQJX\rQli͏XWU˱ͣFYjVDXNKLWUtFKO\ 42
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KӂT QUҦ 9¬%¬1/8ҰN 45
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3.2 ҦQKKѭӣng cӫDFiF\Ӄu tӕ ÿӃQKjPOѭӧng triterpenoid 46
7¬,/,ӊU THAM KHҦO 102
PHӨ LӨC 112
Trang 10DANH MӨ&+Î1+
+uQK1ҩm linh chi 3
+uQKCҩXWU~Fȕ-glucan SKkQOұp tӯ nҩm linh chi [13] 4
+uQK&{QJWKӭc cҩu tҥo cӫa squalen 5
+uQK&iFWULWHUSHQRLGWKXӝFQKyPĮ-amyrin 6
+uQK&iFWULWerpenoid thuӝFQKyPȕ amyrin 7
+uQK&iFWULWHUSHQRLGWKXӝFQKyPlupeol 7
+uQK&ѫFKӃ NKiQJR[\KyDEҵQJFiFKFKRÿLӋn tӱ 9
+uQKҦQKKѭӣng cӫDWULWHUSHQRLGÿӃQTXiWUuQKWӃ EjRFKӃWWKHRFKXNu 10
+uQK&ѫFKӃ NKiQJNKXҭn cӫa triterpenoid 11
+uQK1JX\rQWҳc thӱ nghiӋm hoҥWWtQKNKiQJXQJWKѭ 26
+uQK4X\WUuQKWUtFK ly triterpenoid tӯ nҩm linh chi bҵQJSKѭѫQJSKiSenzyme kӃt hӧSVLrXkP 34
+uQKPhә UV ± YLVYjÿѭӡng cong chuҭn cӫa axit ursolic ӣ ɉ݉ܽݔ ± 548 nm 41
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+uQKҦQKKѭӣQJFӫDQӗQJÿӝHQ]\PHÿӃQKjPOѭӧQJWULWHUSHQRLG 47
+uQKҦQKKѭӣng cӫa tӹ lӋ dung P{LQJX\rQOLӋXÿӃQKjPOѭӧng triterpenoid 48
+uQKҦQKKѭӣng cӫa nhiӋWÿӝ VLrXkPÿӃQKjPOѭӧng triterpenoid 49
+uQKҦQKKѭӣng cӫa thӡLJLDQVLrXkPÿӃQKjPOѭӧng triterpenoid 50
Trang 11+uQKҦQKKѭӣng cӫDF{QJVXҩWVLrXkPÿӃQKjPOѭӧng triterpenoid 51+uQK(a) BiӇXÿӗ SKkQSKӕi nӱa chuҭQYjE %LӇXÿӗ 3DUHWRFiFÿLӇm A, B, C, D, Yj( 55+uQKҦQKKѭӣQJÿӗng thӡi cӫa nӗQJÿӝ enzyme YjWӹ lӋ HWDQROQJX\rQOLӋu 58+uQKҦQKKѭӣQJÿӗng thӡi cӫDF{QJVXҩWVLrXkPYjQӗQJÿӝ enzyme 59+uQKҦQKKѭӣQJÿӗng thӡi cӫa etanol: tӹ lӋ QJX\rQOLӋXYjF{QJVXҩWVLrXkP 60+uQK3 11: Phә FTIR cӫDFDRWUtFK 61+uQK +uQKҧnh SEM cӫa quҧ thӇ nҩPOLQKFKLDE WUѭӟFYjFG