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1 1,2, 3,2, 1, 2, 2 (1., ; 2., ; 3., ) 摘要 : 采用均匀设计从海带中热水提取岩藻聚糖硫酸酯, 以粗多糖提取率 (Y w ) 岩藻糖提取率(Y f ) 和硫酸根提取率 (Y s ) 为评价指标, 并采用综合加权评分法综合考虑各试验因素对以上三个指标的影响利用 Minitab 软件分析加权值 Y 与各因素的相关性, 再通过回归方程优化岩藻聚糖硫酸酯的工艺参数建立了加权值 Y 与提取温度 (X 1 ) 提取时间(X 2 ) 和液料比 (X 3 ) 之间的二次回归模型, 即 Y= x x x x x x 1 x 2 优化后获得最佳提取工艺参数关键词 : 海带 ; 岩藻聚糖硫酸酯 ; 均匀设计 ; 回归方程 ; 热水提取 ; 加权评分中图分类号 : P714.5 文献标识码 : A 文章编号 : (2012) (fucoidan),,, [1] [2] [3-4] [5-7],, L-,,, [2, 5], [8] [9],,, [10-18] [19-22] : [23-24] ; [25] ; [23-24, 26-31],,,, (%)(%) (%) 44 / 2012 / 36 / 11, 1 材料与方法 1.1 材料和仪器 1.1.1, , 95% ; K 2 SO 4 BaCl 2 ; L- : L-(Fuc)(Sigma) ADVENTURER TM, ( ); DF-101, ; SC-3610, ; RE-2000, ; LGJ-12, ; UV-2000, ( ) ; HH-4 : ; : : ( ) : (1987-),,,,, : , qugy2006@ 163.com;,, wangdf@ouc.edu.cn;,, ljhan@qdio.ac.cn

2 1.2 试验方法 1.2.1,,,, 1 Y w : (%)= / 100% Y f : (%)=( )/ 100% Y s : (%)=( )/ 100% [32] : Gibbons L-: y=0.0009x , R 2 = [33] : Dodgson : y=2.6999x ; R 2 = [11, 13, 15, 17, 21, 25], 3 : (X 1 ) (X 2 ) (X 3 ), (Y w )(Y f )(Y s ) 6, 1 1, U12* (12 3 ),, [34-35], D= Minitab,,,, 2 结果与讨论 1 Fig. 1 The flow chart of hot water extracting process of fucoidan from Laminaria japonica 2.1 试验方案及分析结果, (Y w )(Y f ) (Y s ),, 表 1 因素水平表 Tab. 1 The factor-level list for uniform design X1( ) X2 (h) X3(mL/g) Marine Sciences / Vol. 36, No. 11 /

3 ,,,, ( %) (%) (%) 0. 1, 0. 45,0. 45, 100,,, 各实验因素对单个评价指标 Y w Y f 和 Y s 的影响 Minitab 16 (Y w )(Y f )(Y s ),, X 1, X 2, X 3 [35],, 3 表 2 热水提取岩藻聚糖硫酸酯均匀设计试验方案及分析结果 Tab. 2 Conditions and analysis results of hot water extracting process of fucoidan from Laminaria japonica by uniform design X 1 ( ) X 2 (h) X 3 (ml/g) Y w (%) Y f (%) Y s (%) Y 表 3 Y w Y f 和 Y s 的回归方程及其显著性检验 Tab. 3 Results and significance tests of quadratic polynomial stepwise regression of significance tests of Y w Y f and Y s r R 2 F p 1 Y w = x x x 1 x Y f = x x 2 x x Y s = x x x 1 x x x x 1 =X , x 2 =X 2-3.5, x 3 =X P 0.05, =0.05,, Y w P=0.029 Y f Y s, Y w r=0.811, R 2 =0.658,,,, t 4 4 Y w X 1 >X 3 >X 1 X 2, X 1, X 3, X 2, ; 46 / 2012 / 36 / 11

4 Y f X 2 1 > X 2 X 3 >X 1, X 2 1, X 2 X 3, X 2 X 3 ; Y s X 1 >X 2 3 >X 1 X 2 >X 2 2 >X 3, X 1, X 3, X 2 表 4 Y w Y f 和 Y s 偏回归系数的显著性检验 Tab. 4 Significance tests of regression coefficients obtained by the quadratic polynomial regression models of Y w, Y f and Y s t p Y w Y f Y s x x x 1 x x 1 x x 2 x x x x x 1 x x 3 x x 2 x , (X 1 ) (%)(%)(%), (X 3 )(%) (%), (X 2 ),, 2.3 各实验因素对加权评分的影响 Y, : Y= x x x x x x 1 x 2 ; x 1 =X , x 2 =X 2-3.5, x3=x R=0.944; F=6.85, p=0.026<0.05, =0.05,, R 2 =0.892, R 2 =0.762, Y Y w Y f Y s,, t 5 5 Y X 1 > X 2 3 > X 2 1 >X 3 >X 1 X 2 >X 2, X 1, X 3, X 2 表 5 加权值 Y 偏回归系数的显著性检验 Tab. 5 Significance tests of regression coefficients obtained by the quadratic polynomial regression model of Y t P x x 1 x x 3 x x x 1 x x 工艺条件参数优化 Y,, x 1 =25, x 2 =-2.5, x 3 =12.5(X 1 =100, X 2 =1, X 3 =35), 100, 1h, 35(mL:g) 2.5 验证试验, 6.50%, 2.65%, 1.36% Y , 12, , 3 结论, (Y w )(Y f ) Marine Sciences / Vol. 36, No. 11 /

5 (Y s ), : (1) Y Y w Y f Y s (2) Y, 100, 1 h, 35(mL:g) (3), 5.39% 6.50%, 2.65%, 1.36%,,, : [1] Chandía N P, Matsuhiro B. Characterization of a fucoidan from Lessonia vadosa (Phaeophyta) and its anticoagulant and elicitor properties[j]. International Journal of Biological Macromolecules, 2008, 42(3): [2] Wang J, Zhang Q, Zhang Z, et al. Antioxidant activity of sulfated polysaccharide fractions extracted from Laminaria japonica[j]. International Journal of Biological Macromolecules, 2008, 42(2): [3] Synytsya A, Kim W, Kim S, et al. Structure and antitumour activity of fucoidan isolated from sporophyll of Korean brown seaweed Undaria pinnatifida[j]. Carbohydrate Polymers, 2010, 81(1): [4] Ale M T, Maruyama H, Tamauchi H, et al. Fucoidan from Sargassum sp. and Fucus vesiculosus reduces cell viability of lung carcinoma and melanoma cells in vitro and activates natural killer cells in mice in vivo[j]. International Journal of Biological Macromolecules, 2011, 49(3): [5] Sinha S, Astani A, Ghosh T, et al. Polysaccharides from Sargassum tenerrimum: Structural features, chemical modification and anti-viral activity[j]. Phytochemistry, 2010, 71: [6] Cimino P, Bifulco G, Casapullo A, et al. Isolation and NMR characterization of rosacelose, a novel sulfated polysaccharide from the sponge Mixylla rosacea[j]. Carbohydrate Research, 2001, 334: [7] Anastyuk S D, Shevchenko N M, Ermakova S P, et al. Anticancer activity in vitro of a fucoidan from the brown alga Fucus evanescens and its low-molecular fragments, structurally characterized by tandem mass-spectrometry[j]. Carbohydrate Polymers, 2012, 87(1): [8],,,. [J]. ( ), 2002(2): [9],,,. [J]., 2006(4): [10],,,. [J]., 2006(6): [11],,,. [J]., 2003(5): [12],,. [J]., 2004(6): [13],,. [J] (4): [14],,. [J]. ( ). 2006(1): [15],. [J] (2): [16],,. [J] (12): [17],,,. [J] (6): [18],,,. [J] (8): [19],,,. [J] (10): [20],,,. [J] (30): [21],,,. [J] (4): [22],,,. [J] (6): / 2012 / 36 / 11

6 [23],,,. [J] (2): [24],,. [J] (5): [25],,. [J] (7): [26], 慥. [J] (3): [27],. [J] (3): [28],. [J] (3): [29],,. [J] (1): [30],,. [J] (1): [31] 喆,,,. [J] (11): [32] Gibbons M N. The determination of methylpentoses[j]. Analyst, 1955, 80: [33] Dodgson K S, Price R G. A note on the determination of the ester sulphate content of sulphated polysaccharides[j]. Biochem J. 1962: [34]. [M]. :, 1994: [35],. ( 2 )[M]. :, 2010: Study on evaluation model of extracting process of fucoidan from Laminaria japonica QU Gui-yan 1,2, LIU Xu 3,2, WANG Dong-feng 1, YUAN Yi 2, HAN Li-jun 2 (1. Department of Food Science and Engineering, Ocean University of China Qingdao , China; 2. Institute of Oceanology, Chinese Academy of Sciences, Qingdao , China; 3. College of Tea&Food Science and Technology, Anhui Agricultural University, Hefei , China) Received: Jan.,5,2012 Key words: Laminaria japonica; fucoidan; uniform design; regression equation; hot water; weighting values Abstract: Fucoidan was extracted from Laminaria japonica by hot water through uniform design. To optimize the extraction parameters, the weighting values of the separate yields of crude fucoidan (Y w ), fucose (Y f ) and sulfate (Y s ) were used by Minitab. The results showed that the behavior of system was explained by the following quadratic equation between the weighting values (Y) and the factors (extraction temperature, extraction time and the ratio of solution to algae power): Y= x x x x x x 1 x 2. Based on that equation, the optimization condition was that extraction temperature 100, extraction time 1h and the ratio of so- lution to algae power 35 (ml:g). Under this condition, the extraction rate of crude fucoidan, fucose and sulfate group was 6.50%, 2.65% and 1.36%, respectively. ( 本文编辑 : 康亦兼 ) Marine Sciences / Vol. 36, No. 11 /

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g 16 6 2018 11 Chinese Journal of Bioprocess Engineering Vol. 16 No. 6 Nov. 2018 doi 10. 3969 /j. issn. 1672-3678. 2018. 06. 004 1 2 1 1 1. 330200 2. 330006 4 Box-Behnken 1 25 g /ml 121 92 min 2 6. 284%