8 1 2010 1 Chinese Journal of B iop rocess Engineering Vol. 8 No. 1 Jan. 2010 doi: 10. 3969 / j. issn. 1672-3678. 2010. 01. 011 1, 2, 1, 1, 1, 1 (1., 710062; 2., 710062) :, (WCP) (UCP) WCP UCP 871997% 721937% Sephadex G 200 WCP UCP, WCP3 1 UCP3 1 2, ( FTIR) (AFM ) : WCP3 1 UCP3 1, WCP3 1 UCP3 1, C O C O ;WCP3 1, UCP3 1,, : ; ; : Q336 : A : 1672-3678 (2010) 01-0056 - 05 Effect of ultra son ic2a ssisted extraction on polysacchar ide structure from C oprinus com a tus character ized by FT IR and AFM L IU Na2n 1, ZHANG J ing 2, L IDai 1, XU Ping2p ing 1, WANG Bo 1, SUN Run2guang 1 (1. Laboratory of Biophysics and B iomedical Technology, College of Physics and Information Technology, Shaanxi Normal University, Xiπan 710062, China; 2. College of Food Engineering and Nutrition Science, Shaanxi Normal University, Xiπan 710062, China) Abstract: Polysaccharides were extracted from Coprinus com a tus by boiling water p rocess and ultrasonic2 assisted p rocess, the obtained polysaccharides were designated aswcp and UCP, respectively. The total sugars of WCP and UCP were 891997% and 721937% determ ined by phenol2sulfuric acid m ethod. The crude polysaccharides were purified by CTAB and Sephadex G2200 in sequence, and the obtained ones were designated aswcp321 and UCP321. FTIR showed that there existed 2pyranoside in the polysaccha2 rides and AFM showed thatwcp321 on m ica existed in sp iral and spherical form, but UCP321 existed in spherical form. The ultrasonic collap se of cavitation bubbles generated intense shock waves that damaged the intrachain or interchain hydrogen bonds. Key words: Coprinus com a tus; ultrasonic extraction; polysaccharide : 2009-04 - 20 : (10874108) ; ( SJ08A16) : (1985 ),,,, : ; ( ),,, E2mail: sunrun2 guang@ snnu. edu. cn
1 : 57 (Coprinus com atus),, [ 1-2 ], 3 16, [ 3 ],, [ 4 ],,,,, ( FTIR ) (AFM ) 2, 2, 1 1. 1 ; H 2 SO 4 NaOH (CTAB ) NaCl, 1. 2 KQ3200 ( ), JY92 ), HPX ( 9162MBE ( ), CF16RX ( ), TU 1810 ( ), EQU INX55 ( B rucher ), A lphal 4 ( Christ ), SPM 9500J3 ( ) 1. 3 1. 3. 1 : 4 80% (, ) (20 m in 3 ) 10 15 m in ( 10 s 90 15 s, 300 W ), 3 ( 4 000 r/m in 10 m in ) 3 d (UCP) [ 5 ] 1. 3. 2 : 4 80% ( 20 m in 3 ) 10 215 h, 3 ( 4 000 r/m in 10 m in ) 3 d (WCP) [ 6 ] 2 2. 1 WCP UCP, WCP : 199100 nm, A 199 = 11747; UCP : 205100 nm, A 205 = 11922; 2, 260 nm 280 nm, [ 5 ], WCP UCP [ 7 ] 2. 1. 1 1 ) 105 20 mg, 100 ml,, 012 mg/ml 2) 5 g,, 100 ml, 3) 715 1510 2215 3010 3715 ml, 6 50 ml ( 1 0 ml, ), 0 0103 0106 0109 0112 0115 mg/ml 6 012 ml 014 ml, H 2 SO 4 2 ml, 30 m in, 488 nm (A 488 ) ( ), (A 488 ),
58 8 A 488 = 01005 4 + 01000 2 R 2 = 01999 8 2. 1. 2 WCP UCP 01050 g, 50 ml, 1 mg/ml 1 ml 25 ml 0104 mg/ml 012 ml 2. 1. 1, WCP3 UCP3, 1, WCP UCP 871997% 721937%, UCP WCP,, 2. 2 2. 2. 1 WCP UCP, 1) 5 g WCP UCP 40 ml, 5% (CTAB ) 50 ml,, 12 h 4 000 r/m in 4 20 m in 10% NaCl, 3 95%,, 4 000 r/m in 20 m in 3 d ( 8 000 ),, WCP1 ( 01986 g) UCP1 (01597 g) 2) CTAB 1% H 3 BO 3 ph 610, 2 mol/l NaOH ph 1110,, 4 000 r/m in 4 20 m in, 10% NaCl, 2% ph 710, 3 95%,, 3 d, UCP2 (01335 g) WCP 3), ph 414, 3 95%, 4 24 h, 4 000 r/m in 4 20 m in,, 3 d, WCP3 (21029 g) UCP3 (21007 g) 2. 2. 2 WCP3 UCP3 WCP3 UCP3 Sephadex G 200 WCP3 1 UCP3 1,, WCP3 UCP3 2. 3 WCP3 1 UCP3 1 WCP3 1 UCP3 1, KB r,,, 4 000 400 cm - 1 WCP3 1 UCP3 1 : 3 406 cm - 1 3 409 cm - 1 O H, ; 2 929 cm - 1 C H, ; [ 8 ], 1 634 1 631 cm - 1 N H, CH 3 CONH I, ; 1 385 cm - 1 1 388 cm - 1 C H, ; [ 9 ], 1 250 950 cm - 1 ; [ 10-11 ], 930 cm - 1, 840 cm - 1,, WCP3 1 UCP3 1 UCP3 1 1 420 cm - 1 C O, WCP3 1, UCP3 WCP3 1 C O, C 1 C O : O C O, 2. 4 WCP3 1 UCP3 1 WCP3 1 UCP3 1 10 g/ml, 015 L,, 20 m in SPM 9500J3
1 : 59 1 Sephadex G 200 W CP3 UCP3 F ig. 1 Sephadex G2200 column chroma tography of W CP3 and UCP3 [ 12-13 ], 2 2 W CP3 1 UCP3 1 ( ) F ig. 2 AFM images of W CP321 and UCP321 2 : WCP3 1 UCP3 1, (2150 m 2150 m ) : WCP3 1, ; UCP3-1 2 ( a) WCP3 1,,, ; 2 ( b) UCP3 1,,, 3 1), C O C O, COOH, 2),,,, : [ 1 ],,,. [ J ]., 2005, 26 (8) : 1612163. W u Q iaofeng, L iu Jingjuan, Chen Jing, et al. Nutrient analysis of Coprinus com atus[ J ]. Food Nutrition, 2005, 26 (8) : 1612163. [ 2 ],,,. [ J ]. :, 2009, 24 (1) : 45249. Yu Jie, Cui Shichao, Song Caixia, et al. Purification and spectra a2 nalysis of polysaccharide from the sporophore of Coprinus com atus [ J ]. J Shantou Univ: Natural Science, 2009, 24 (1) : 45249.
60 8 [ 3 ],,,. [ J ]., 2007, 13 (20) : 84286. Yu Jie, Cui Pengju, Chen Meizhen, et al. Progress of researches on polysaccharide [ J ]. Anhui Agric Sci Bull, 2007, 13 (20) : 84286. [ 4 ],,. [ J ]., 2007, 35 (13) : 383823839; 3855. Deng Gongcheng, Chen Jiangtao, Gao L iπan. Op tim ization of fer2 mentation conditions of Coprinus com atus [ J ]. Anhui Agric Sci, 2007, 35 (13) : 383823839; 3855. [ 5 ] Zhang Mei, Zhang L ina, Keung Cheung Chi Peter, et al. Molecular weight and anti2tumor activity of the water2soluble polysaccharides isolated by hotwater and ultrasonic treatment from the sclerotia and mycelia of P leurotus tuber2regium [ J ]. Carbohydrate Polymers, 2004, 56 (2) : 1232128. [ 6 ],,,. [ J ]., 2007, 34 (6) : 107121076. Yao Yujing, Yang Renzhi, Zhang Jingsong, et al. Optimum isolation and structural analysis of polysaccharide from the fruiting bodies of Coprinus com atus[ J ]. M icrobiology, 2007, 34 (6) : 107121076. [ 7 ],,,. RCPS [ J ]., 2008, 28 (3) : 6422644. Song Xuewei, Ren Lei, Han Yongp ing, et al. Purification and com2 position analysis of polysaccharide RCPS from R hodiola crenulata [ J ]. Spectroscopy and Spectral Analysis, 2008, 28 (3) : 6422644. [ 8 ]. [M ]. 2. :, 1999: 11; 1122120; 59264; 1932198. [ 9 ],,,. [ J ]., 2007, 30 (2) : 1022105. L iu W ei, Yu Yuanhua,Mao Yajie, et al. Analysis on isolation, puri2 fication, component and structure of A rm illarialuteo2virens [ J ]. Changchun Univ Sci Tech, 2007, 30 (2) : 1022105. [ 10 ],,,. 2225 [ J ]., 2008, 29 (4) : 2672271. L iw eixin, Chen Q ian, L i Pinglan, et al. Structural study on exopo2 lysaccharide produced by B ifidobacterium 2225 [ J ]. Food Science, 2008, 29 (4) : 2672271. [ 11 ],,. [ J ]., 2007, 29 (2) : 15217; 35. W ang Yue, Chen Aπmei, L i B ingchao. Purification and analysis of water soluble polysaccharides from root, stem and leaf of A ralia continentalis[ J ]. Am ino Acids & B iotic Resources, 2007, 29 ( 2) : 15217; 35. [ 12 ],. [ J ]., 2006, 64 (24) : 246722472. Sun Runguang, Zhang Jing. A study of helical structure of glycyr2 rhiza polysaccharides by atom ic force m icroscope[ J ]. Acta Chim ica Sinica, 2006, 64 (24) : 246722472. [ 13 ],,,. [ J ]., 2007, 16 ( 6 ) : 7492753. Dai L ing, L iao Hongmei, Shen Yeshou, et al. Obervation of struc2 ture morphology and self2assembly of polysaccharide PSM2b2A by atom ic force m icroscopy[ J ]. Acta Laser B iology Sinica, 2007, 16 (6) : 7492753.,,, 2009 11, ;, ;,,, 12,,, 3,,,,,, ( )