Optimization of Extracting Technology of Collagen from Fish Scale

Transcription

1 , Vol. 27, No , * ( ) 10% 1% 3 48h SDS-PAGE SDS-PAGE Optimization of Extracting Technology of Collagen from Fish Scale ZHONG Zhao-hui 1 LI Chun-mei 1, *LIANG Jin-e 2 GU Hai-feng 1 DOU Hong-liang 1 (1.College of Food Science and Technology, Huazhong Agricultural University, Wuhan , China 2.Food Industry Association of China, Beijing , China) Abstract Collagens were extracted from fish scale of grass carp with pepsin and the effects of extraction media, pretreating, stirring, extract times and time on the yield of collagen were studied. The results indicated that the optimum extracted conditions are 1% citric acid as extraction solutions with a magnetic stirrer during the whole process, being extracted three times and 48 hours each time after removing impurity by microwave and decalcifing with 10% citric acid. Purified collagens were analyzed by SDS-PAGE and FT-IR, compared with calf tendon type collagen. The results confirmed that the extracted collagens are typical typecollagen and of electrophoretic purity. Key wordsfish scalecollagentechnologysds-pageft-ir * (1978-) Table 5 5 (xsd,n=3) The comaprison of different extraction techniques(xsd,n=3) (%) 30% 2.0h : %pH h 1:15 ph % h 1:15 [1]. [M]. :, [2],,,. [J]. 3.1, 1997, 12(4): [3],,,., [J]., 1999, 14(2): [4],. [J].,, 2003, 30(2): [5],,. [J]., 2004, 25(1): [6],,. [J]. 3.2, 2005, 26(1):

2 2006, Vol. 27, No S912 A (2006) % 200ml 0.6mol/L 200ml 24h 50%70% 0.1mol/L 100ml 1% 100ml, 25%55% 150ml [1] 2min 30min 10% 200ml 24h Kimura, S. [2] 0.5mol/L 10ml 1% 100ml %5% g 1% (1% (W/V)) ( 2 48h) 1 10% NaCl 48h 0.6mol/L HCl 24h 1.1 2min 40 30min 10% 24h ( 1:3000) Fluka Sigma 2min R-250 Sigma 30min 0.1mol/L NaOH 48h 10% 24h mol/L NaOH 48h 0.1mol/L NaOH 24h 1%(W/V) 0.6mol/L HCl 24h g 3 ph 23 ( 0.01mol/L 0.5mol/L 1% 2 48h) 10.00g ( g 3 48h)

3 , Vol. 27, No h 3 48h 2 2 Table 2 Effect of pretreating on the yield of collagen 48h 3 (g) r/min 20min % 0.9mol/L 4500r/min 20min 0.5mol/L 1 [3][6] 10% 3d 0.9mol/L mol/L SDS-PAGE 2.3 SDS-PAGE 7.5% 5% Tris- (ph % SDS) 0.05mol/L ph8.0 Tris-HCl (2% SDS5% -ME10% 0.02% ) 0.25% (7.5% 5% ) [79] 0.3 R mg 1ml 100 5min10000r/min 20min 1 100V Fig.1 Effect of stirring on the yield of collagen 160V 3 4h KBr Nicolet- 2.4 SX cm cm mol/L 0.5mol/L h, 2 48h, 2 48h, 3 1% 0.01mol/L 0.5mol/L 1% 2 Fig.2 Effect of extract times and time on the yield of collagen 1 Table 1 The choice of extraction medium 2 48h 3 138% 35% 0.01mol/L1% 0.5mol/L1% (g) (1 2) 41%55% 38%46% 1 2, 3 [2][10,11] SDS-PAGE (3) 3 (g) (g)

4 2006, Vol. 27, No ( 211.1kD) 1(3)( kD) 2 ( 107.2kD) I (cm -1 ) 200kD 130kD 97.4kD 66.2kD 43kD ( 3) SDS-PAGE Fig.3 SDS-PAGE electrophoresis of fish scale collagen FT-IR 4 Doyle [12] N-H cm -1 4 A 3323cm -1 N-H 3084cm -1 B 1d C-N 3d cm -1 C=O cm cm [13] cm -1 10% C-N N-H 1% 0.01mol/L 1552cm mol/L 3 48h (Gly-Pro-Hyp)n 4.2 SDS-PAGE FT-IR cm cm -1 C-N N-H Gly Pro CH2 [14] 1238cm -1 N-H [1],,. [J]., 2000, Hyp 21(3): (%) Amide A Amide B Amide I Amide II Amide III Fig.4 FT-IR spectra 1203cm cm

5 , Vol. 27, No. 07 ( ) Study on Application of Radiation Processing Technology on Food Production ZHU JunYANG Zong-quSONG Wei-dongZHAO Hui-dongZHOU JieWANG Yun ZHAO Mei-hongQUAN Yan-jun (Isotope Institute, Henan Academy of Sciences, Zhengzhou , China) Abstract Application of radiation processing technology on food production was studied with a series of soybean protein products. It showed that physical and chemical indices and functional indices of soybean protein products are all improved after treatment of radiation processing technology. Furthermore, radiation processing technology raises hygienic quality of products significantly. The quality indices of irradiated products conform to the Enterprise Standards of Product Quality. The development and application of radiation processing technology have solved a problem of microbe contamination that has perplexed soybean protein production over a long period of time, so radiation processing ensures safety for industrialization of soybean protein production. Key words radiation processing technology soybean protein physical and chemical indices functional indices TS205.9 A (2006) (1971-) [2] Kimura S, Zhu X, Matsui R, et al. Characterisation of fish muscle type I collagen[j]. Journal of Food Science, 1988, 23: [3] Ogawa M, Portier R J, Moody M W, et al. Biochemical properties of bone and scale collagens isolated from the subtropical fish black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus) [J]. Food Chemistry, 2004, 88: [4] Nagai T, Suzuki N. Preparation and characterization of several fish bone collagen[j]. Journal of Food Biochemistry, 2000a, 24: [5] Nagai T, Suzuki N. Isolation of collagen from fish waste material-skin, bone and fins[j]. Food Chemistry, 2000b, 68, [6] Nomura Y, Sakai H, Ishii Y, et al. Preparation and some properties of type I collagen from fish scales[j]. Bioscience Biotechnology and Biochemistry, 1996, 60: [7]. [M]. :, [8],. [M]. :, [9],. [M]. :, [10] Saito M, Takenouchi Y, Kunisaki Y, et al. Complete primary structure of rainbow trout type I collagen consisting of alpha1 (I)alpha2(I) alpha3(i) heterotrimers[j]. European Journal of Biochemistry, 2001, 268: [11] Yoshida C, Fujisawa S, Mizuta S, et al. Identification and characterisation of molecular species of collagen in fish[j]. Journal of Food Science, 2001, 66: [12] Doyle B B. Infrared spectroscopy of collagen and collagne-like polypeptides [J]. Biopolymers, 1975, 245: [13] Myung Chul Chang, Junzo Tanaka. FT-IR study for hydroxyapatite/ collagen nanocomposite cross-linked by glutaraldehyde[j]. Biomaterials, 2002, 23: [14] He Li, Hua Lin Chen, Rong Luo, et al. The interaction between collagen and an aluminum tanning agent[j]. Macromol Bioscience, 2003, (3):