酶法改性魔芋葡甘聚糖强化海藻酸—壳聚糖微囊缓释性能研究
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摘要
本文探讨了β-甘露聚糖酶水解魔芋葡甘聚糖（KGM）的过程；研究了将不同时间水解产物用作海藻酸(ALG)-壳聚糖(CHI)体系添加剂，缓释牛血清白蛋白（BSA）与尼莫地平固体分散体的过程，探讨了不同水解时间魔芋葡甘聚糖与海藻酸、壳聚糖的相互作用。试验结果如下：
在酶活200U/l，底物浓度小于10mg/ml下，测得米氏常数Km=8.27 mg/ml，最大反应速率Vmax=0.0221 mg/ml.min；在酶活225U/l，底物为10mg/ml，水解温度40℃下，反应初期粘度迅速下降，而还原糖变化较小，约1小时后反应停止。色谱分析表明，魔芋葡甘聚糖的部分水解产物的分子量分布呈近似正态分布，水解2min与水解5min的魔芋葡甘聚糖的平均分子量分别为1500000与700000左右。
粘度测试表明，海藻酸与魔芋葡甘聚糖间有明显的增稠作用；红外测试表明，水解魔芋葡甘聚糖与海藻酸、壳聚糖的相互作用相对较强，魔芋葡甘聚糖的水解程度可用红外谱图表征；扫描电镜（SEM）观察表明，水解5min魔芋葡甘聚糖微囊内无相分离，微囊表面覆壳聚糖量较大；溶胀度测试表明，微囊在模拟肠液中有明显的溶胀度延迟现象，覆壳聚糖膜同时添加魔芋葡甘聚糖的体系，在模拟肠液中的溶胀度明显减少，有利于药物的缓释。
海藻酸-酶解魔芋葡甘聚糖-壳聚糖微囊在模拟胃肠液中释放蛋白质的实验表明。适当降低魔芋葡甘聚糖分子量，增大魔芋葡甘聚糖浓度，并选择较高的壳聚糖分子量和壳聚糖浓度，同时适当降低凝胶离子浓度将会获得较好的缓释效果。在模拟肠液中，缓释效果最佳的体系为：ALG浓度2%w/v；KGM浓度为0.5% w/v，水解5min；CHI浓度为0.5%w/w，分子量为1.4110×106；凝胶离子浓度[Ca2+]=0.2M。海藻酸-酶解魔芋葡甘聚糖-壳聚糖微囊在模拟胃肠液中释放尼莫地平固体分散体的实验也表明，适当降低魔芋葡甘聚糖的分子量与粘度，尼莫地平在模拟胃、肠液中的释药速率相对较低。
The process of konjac glucomannan (KGM) hydrolysis by β-mannanase was studied in this thesis. Enhancement of controlled-release for alginate (ALG)-chitosan(CHI) microcapsules by enzymatically hydrolyzed glucomannan(EHG) was investigeted. Bull serum albumin (BSA) and nimodipine solid dispersion were used as a model protein for in vitro assessment. The interaction among enzymatically hydrolyzed glucomannan and alginate and chitosan was also discussed.
The experiment of konjac glucomannan hydrolyzed by β-mannanase indicated that the Michaelis constant (Km )and the maximum reaction rate were 8.27 mg/ml and 0.0221 mg/ml.min respectively under enzymatic activity 200U/l. Dynamic viscosity decreased rapidly but reducing sugar changed a little during initial reaction. The reaction stopped after 1 hour when enzymatic activity 225U/l and substrate 10mg/ml hydrolysis temperature 40℃. Chromatographic analysis demonstrated that the molecular weight distribution of EHG was approximately normal distribution, and the average molecular of KGM hydrolyzed for 2min and 5min were about 1500000 and 700000 respectively.
The mixture of ALG and KEM had higher viscosity. There was relatively strong interaction between ALG and EHG and CHI by the graph of infrared (IR). The process of KGM hydrolysis by β-mannanase could be expressed by IR. The scanning electron microscopy (SEM) micrograph indicated that non phase separation exists between ALG and EHG (5min) within microcapsules and the quantity of CHI was higher on the surface of microcapsules. Studies of water of hydration had shown that the swelling of ALG-EHG-CHI microcapsules was lower. Then the usage of EHG helps to enhance the controlled release.
In imitated gastric and intestinal juice, the experiment of BSA release from ALG-EHG-CHI microcapsules indicated that the release rate become slowly with suitably decreasing the molecular weight of KGM and the concentration of gel ions. Raising the concentration of CHI and that of KGM and the molecular weight of CHI also enhance the controlled-release. The better system was 2% ALG and 0.5% EHG of 5min and 0.5% CHI of 1.4110×106 molecular weight. The experiment of nimodipine release from ALG-EHG-CHI microcapsules also demonstrated that proper decrease in the molecular weight of KGM would help to induce the release rate.
在酶活200U/l，底物浓度小于10mg/ml下，测得米氏常数Km=8.27 mg/ml，最大反应速率Vmax=0.0221 mg/ml.min；在酶活225U/l，底物为10mg/ml，水解温度40℃下，反应初期粘度迅速下降，而还原糖变化较小，约1小时后反应停止。色谱分析表明，魔芋葡甘聚糖的部分水解产物的分子量分布呈近似正态分布，水解2min与水解5min的魔芋葡甘聚糖的平均分子量分别为1500000与700000左右。
粘度测试表明，海藻酸与魔芋葡甘聚糖间有明显的增稠作用；红外测试表明，水解魔芋葡甘聚糖与海藻酸、壳聚糖的相互作用相对较强，魔芋葡甘聚糖的水解程度可用红外谱图表征；扫描电镜（SEM）观察表明，水解5min魔芋葡甘聚糖微囊内无相分离，微囊表面覆壳聚糖量较大；溶胀度测试表明，微囊在模拟肠液中有明显的溶胀度延迟现象，覆壳聚糖膜同时添加魔芋葡甘聚糖的体系，在模拟肠液中的溶胀度明显减少，有利于药物的缓释。
海藻酸-酶解魔芋葡甘聚糖-壳聚糖微囊在模拟胃肠液中释放蛋白质的实验表明。适当降低魔芋葡甘聚糖分子量，增大魔芋葡甘聚糖浓度，并选择较高的壳聚糖分子量和壳聚糖浓度，同时适当降低凝胶离子浓度将会获得较好的缓释效果。在模拟肠液中，缓释效果最佳的体系为：ALG浓度2%w/v；KGM浓度为0.5% w/v，水解5min；CHI浓度为0.5%w/w，分子量为1.4110×106；凝胶离子浓度[Ca2+]=0.2M。海藻酸-酶解魔芋葡甘聚糖-壳聚糖微囊在模拟胃肠液中释放尼莫地平固体分散体的实验也表明，适当降低魔芋葡甘聚糖的分子量与粘度，尼莫地平在模拟胃、肠液中的释药速率相对较低。
The process of konjac glucomannan (KGM) hydrolysis by β-mannanase was studied in this thesis. Enhancement of controlled-release for alginate (ALG)-chitosan(CHI) microcapsules by enzymatically hydrolyzed glucomannan(EHG) was investigeted. Bull serum albumin (BSA) and nimodipine solid dispersion were used as a model protein for in vitro assessment. The interaction among enzymatically hydrolyzed glucomannan and alginate and chitosan was also discussed.
The experiment of konjac glucomannan hydrolyzed by β-mannanase indicated that the Michaelis constant (Km )and the maximum reaction rate were 8.27 mg/ml and 0.0221 mg/ml.min respectively under enzymatic activity 200U/l. Dynamic viscosity decreased rapidly but reducing sugar changed a little during initial reaction. The reaction stopped after 1 hour when enzymatic activity 225U/l and substrate 10mg/ml hydrolysis temperature 40℃. Chromatographic analysis demonstrated that the molecular weight distribution of EHG was approximately normal distribution, and the average molecular of KGM hydrolyzed for 2min and 5min were about 1500000 and 700000 respectively.
The mixture of ALG and KEM had higher viscosity. There was relatively strong interaction between ALG and EHG and CHI by the graph of infrared (IR). The process of KGM hydrolysis by β-mannanase could be expressed by IR. The scanning electron microscopy (SEM) micrograph indicated that non phase separation exists between ALG and EHG (5min) within microcapsules and the quantity of CHI was higher on the surface of microcapsules. Studies of water of hydration had shown that the swelling of ALG-EHG-CHI microcapsules was lower. Then the usage of EHG helps to enhance the controlled release.
In imitated gastric and intestinal juice, the experiment of BSA release from ALG-EHG-CHI microcapsules indicated that the release rate become slowly with suitably decreasing the molecular weight of KGM and the concentration of gel ions. Raising the concentration of CHI and that of KGM and the molecular weight of CHI also enhance the controlled-release. The better system was 2% ALG and 0.5% EHG of 5min and 0.5% CHI of 1.4110×106 molecular weight. The experiment of nimodipine release from ALG-EHG-CHI microcapsules also demonstrated that proper decrease in the molecular weight of KGM would help to induce the release rate.
引文
冯子玉.葡甘聚糖降血脂及血压的疗效.新药与临床，1997，16（2）：105—106
沈悦玉，张明春，王吰.魔芋胶的结构及其流变性.天津商学院学报，1995，3：17—22
陈运中.魔芋精粉与黄原胶的协同增效作用及应用研究.食品科学，1999，9：12—14
4. 王铭和.唐湛祥.卡拉胶一魔芋粉的协合作用研究.湛江海洋大学学报，2000,
20(2)：34—35
Tye, R.J. Properties and rheology of konjac and carrageenan systems，Polymeric Materials Science and Engineering. Proceedings of the ACS Division of Polymeric Materials Science and Engineering，1990（63）：229—232
莫卫平，蒙义文，贾成禹，等. 葡甘聚糖凝胶及其衍生物研究（Ⅰ）.离子交换与吸附，1992，8（1）：5—9
杨新亭，王林风，王香东.黄原胶与魔芋胶的协效凝胶性研究.食品科学，2001，22（3）：38—40
Noriko Kishida.; Satoshi okjmasn. Preparation of water-soluble methyl konjac glucomannan. Agric Biol Chem, 1978,42(3)：669—670
Kohyama K, In: Gums and Stabilizers of the Food Industry 5, Oxford: IRL Press, 1990
吴贤聪，梁存钧，郭森炎。魔芋葡甘聚糖的提取、鉴定及应用的研究.
李斌，廖勇城，谢笔钧.高效液相色谱法测定魔芋中魔芋葡甘聚糖含量.农牧产品开发.1999,8
12. Vipul Dave,Mihir Sheth,Stephen P. Mccarthy,Jo Ann Ratto and David L. Kapian.Liquid crystalline, rheological and thermal properties of konjac glucomannan . polymer Vol. 39 No.5,pp.1139-11148,1998
13. 沈悦玉, 杨湘庆. 魔芋胶的特征和魔芋凝胶食品. 食品科学, 1995, 16(6): 15- 19.
14. 王照利, 吴万兴, 李科友. 魔芋精粉中甘露聚糖含量测定研究. 食品科学, 1998, 19(3): 56-58.
15. K.Nishinnari ,Yoh Sano ,Kaoru Kohyama. A mixed systerm composed of different molecular weights konjac glucomannan and κ–carrageenan .Ⅱ.Molecular weight dependence of viscoelasticity and thermal properties. Food Hydrocolloids.Vol. 10 no. 2 pp. 229-238,1996
16. K.Nishinnari ,Hiroki Iida ,Kaoru Kohyama. A mixed systerm composed of different molecular weights konjac glucomannan and –carrageenan :large deformation and dynamic viscoelastic study. Food Hydrocolloids.Vol. 7 no. 2 pp. 213-226,1993
庞杰.六偏磷酸钠对魔芋葡甘聚糖干法改性研究.西南农业大学学报，2000年，22（1）：59—61
18. 崔汉钧.魔芋葡甘聚糖固定化环糊精葡基转移酶的研究.天然产物研究与开 发，1993，5（3）：48—54
C.Perols.; B.Piffaut.; J.Scher et al. The potential of enzyme entrapment in konjac cold-melting gel beads. Enzyme and Microbial Technology,1997 20:57—60
周立，蒋磊，郑远旗.魔芋葡苷聚糖凝胶为亲和导析载体与Sepharose 4B的比较研究，Ⅰ.KGM金属螯和层析分离纯化猪血SOD.天然产物研究与开发,1998,10(4):1998,10 (4)：55—58
Wakita M. 公开特许公报（日），昭61—19238（1991,434,354）
Tomoda T. 公开特许公报（日），昭61—21066 （1990,02,212,763）
23 Okaiyi M公开特许公报（日），昭61—19243(1989,434,436)
24. Nakano, Masahiro；Takikawa, Kaori；Arita, Takaichi. Release characteristics of dispersed in konjac gels. Journal of Biomedical Materials Research,1979,13 (5): 811—819
25. Xie, S.S. ; Liu, K.J.; Zhang, Y.X.; Zhang, J.L. et al. Preparation and release test of rotundine-PIC polysaccharide granules. Journal of Macromolecular Science - Pure and Applied Chemistry, 1992,29(10) : 931—938
26. Fanutti C, Ponyi T, Black GW, et al. J Biol Chem, 1995, 270: 29314-29322.
27. 吴襟. β-1,4-D-甘露聚糖酶的纯化和性质[学位论文]. 北京：中科院微生物所, 1997.
AkinoT, Nakamura N, Horikoshi K. β-mannosidase and β-mannanase by an alkalophilic Bacillus sp. Appl.Microbiol.Biotechnol, 1987, 26(4): 323-327.
Firantas S G A, Venzohinskene J I, Paulokonis A B. Prikl.Biokhim.Mikrobiol, 1982, 18: 581-587.
Bruce D, Henk W M, Hilhorst J, et al. Phytochemistry, 1994, 36(4): 829-835.
Barry V, Mccleary. The fine structures of carob and guar galactomannans. Carbohydr Res, 1985, 139: 237-260.
张迎庆，干信，邹群，马丽. 魔芋葡甘低聚糖硫酸酯化衍生物的制备及结构分析. 药物生物技术. 2001,8(4):200-203
M. Yoshimura, K. Nishinari. Dynamic viscoelastic study on the gelation of konjac glucomannan with different molecular weights. Food Hydrocolloids. 13(1999) 227-233
H.Zhang ,M. Yoshimura , K. Nishinari ,M. A. K. Williams ,T. J. Foster , I. T. Norton. Gelation behaviour of konjac glucomannan with different molecular weights.Biopolymers. Vol. 59,38-50(2001)
Mumper R J, Hoffman A S, Puolakkainen P et. al., Calcium-alginate beads for the oral delivery of transforming growth factor-β1: Stabilization of TGF-β1 by the addition of polyacrylic acid within acid-treated beads, J. Control Release, 1994, 30: 241-251
Park K, Robinson J R, Bioadhesive polyners as platforms for oral-controlled drug delivery: Method to study bioadhesion, Int. J. Pharm., 1984,19: 107-127
Chickering D E, Mathiowitz E, Bioadhesive microspheres:ⅠA novel electrobalance-based method to study adhesive interactions between individual microspheres and intestinal mucosa, J. Control. Release, 1995, 34:251-261
Martinsen A, Alginate as immobilization material: Ⅲ. Diffusion properties, Biotechnol. Bioeng., 1992, 39: 186-194
Quong D, Neufeld R J, Skjak-Braek G and Poncelet D, External versus internal source of calcium during the gelation of alginate beads for DNA encapsulation, Biotechnol. Bioeng.,1998, 57: 438-446
Huguet M L, Groboillot A, Poncelet D et. al., Hemoglobin encapsulation in chitosan/calcium alginate beads, J. Appl. Polym. Sci., 1994, 51:1427-1432
Yoshikawa Y, Komuta Y, Nishihara T et. al., Preparation and evaluation of once-a-day injectable microspheres of interferon alpha in rats. J. Drug Target, 1999, 6(6): 449-461
Cho NH, Seong SY, Chun KH et. al., Novel mucosal immunization with polysaccharide-protein conjugates entrapped in alginate microspheres. J. Controlled Release, 1998 ,53(1-3):215-224
Esquisabel A, Hernandez RM, Igartua M et. al., Production of BCG alginate-PLL microcapsules by emulsification/internal gelation, J. Microencapsul, 1997, 14(5): 627-638
Kim HJ, Lee HC, Oh JS et. al., Polyelectroylte complex composed of chitosan and sodium alginate for wound dressing application, J Biomater Sci. Polym. Ed., 1999, 10(5): 543-556
Liu P, Krishnan TR, Alginate-pectin-poly-L-lysine particulate as a potential controlled release formulation, J Pham. Pharmacol., 1999(2): 141-149
Shiraishi S, Imai T, Otagiri M, Controlled-release preparation of indomethacin using calcium alginate gel, Biol. Pharm. Bull, 1993 ,16(11):1164-1168
Takka S, Acarturk F, Calcium alginate microparticles for oral administration: I: Effect of sodium alginate type on drug release and drug entrapment efficiency, J. Microencapsul, 1999, 16(3): 275-290
Tateshita K, Sugawara S, Imai T et. al., Preparation and evaluation of a controlled-release formulation of nifedipine using alginate gel beads, Biol. Pharm. Bull, 1993, 16(4): 420-424
Aslani P, Kennedy RA, Effect of gelation condition and dissolution media on the release of paracetemal from alginate beads, J. Microencapsul, 1996, 13(5):601-614
Sugawara S, Imai T, Otagiri M, The controlled release of prednisolone using alginate gel, Pharm. Res., 1994, 11(2): 272-277
Hari PR, Chandy T, Sharma CP, Chitosan/calcium alginate microcapsules for intestinal delivery of nitrofurantoin, J. Microencapsul, 1996, 13(3):319-329
Pillay V, Fassihi R, In vitro release modulation from crosslinked pellets for site-specific drug delivery to the gastrointestinal tract. I. Comparison of pH-responsive drug release and associated kinetics, J. Controlled Release, 1999, 59(2): 229-242
Sezer AD, Akbuga J, Release characteristics of chitosan treated alginate beads: II. Sustained release of a low molecular drug from chitosan treated alginate beads, J. Microencapsul, 1999 ,16(6): 687-696
Knoll A, Schmidt S, Chapman M et. al., A comparison of two controlled-release delivery systems for the delivery of amiloride to control angiogenesis, Microvasc. Res., 1999 , 58(1):1-9
Fathy M, Safwat SM, el-Shanawany SM et. al., Preparation and evaluation of beads made of different calcium alginate compositions for oral sustained release of tiaramide, Pharm. Dev. Technol., 1998 , 3(3): 355-364
Prisant LM, Bottini B, DiPiro JT et. al., Novel drug-delivery systems for hypertension, Am. J. Med., 1992 , 93(2A): 45S-55S
Kuen Yong L, Won Ho P, Wan Shik H, Polyelectrolyte complexes of sodium alginate with chitosan or its derivatives for microcapasules, J. Appl. Polym. Sci.,
1996, 63: 425-432
Kaneko K, Kanada K, Miyagi M et. al., Formation of water-insoluble gel in dry-coated tablets for the controlled release of theophylline, Chem. Pharm. Bull (Tokyo) , 1998, 46(4): 728-729
Yomota C, Miyazaki T, Okada S, Sustained-release effect of the direct compressed tablet based on chitosan and Na alginate, Yakugaku Zasshi, 1994, 114(4): 257-263
Czarnecki W, Wielgus S, Factors affecting the release of papaverine from spheres, Acta. Pol. Pharm., 1996, 53(5): 331-336
Kishi K, Sonomura T, Nishida N et.al., Alginate gel beads for chemoembolization: initial report, Nippon Igaku Hoshasen Gakkai Zasshi, 1995 , 55(5):300-304
Whitehead L, Fell JT, Collett JH et. al., Floating dosage forms: an in vivo study demonstrating prolonged gastric retention, J. Controlled Release, 1998,55(1):3-12
L Rowsen Moese, Dileep K L, Chandra P Sharma, Bete Cyclodextrin-insulin-encapsulted Chitosan/alginate matrix: for oral delivery of insulin, J. Appl. Polym. Sci., 2000, 75: 1089-1096
曹海辉，王亦农，马建标等，肽类药物口服剂型材料及控制释放性能研究Ⅱ.含有添加剂的壳聚糖-海藻酸盐微囊对胰岛素的控释作用，离子交换与吸附，1999，15（6）：496-503
Gursoy A, Cevik S, Sustained release properties of alginate microspheres and tabletted microspheres of diclofenac sodium, J.Microencapsul, 2000, 17(5): 565-575
Kang Wang and Zhimin He, "Alginate-konjac glucomannan-chitosan beads as controlled release matrix", International Journal of Pharmaceutics, 2002, 244: 117-126
邢琦.尼莫地平缓释微囊的制备及其释放性能的研究. 天津大学硕士论文. 2002
沈悦玉，张明春，王吰.魔芋胶的结构及其流变性.天津商学院学报，1995，3：17—22
陈运中.魔芋精粉与黄原胶的协同增效作用及应用研究.食品科学，1999，9：12—14
4. 王铭和.唐湛祥.卡拉胶一魔芋粉的协合作用研究.湛江海洋大学学报，2000,
20(2)：34—35
Tye, R.J. Properties and rheology of konjac and carrageenan systems，Polymeric Materials Science and Engineering. Proceedings of the ACS Division of Polymeric Materials Science and Engineering，1990（63）：229—232
莫卫平，蒙义文，贾成禹，等. 葡甘聚糖凝胶及其衍生物研究（Ⅰ）.离子交换与吸附，1992，8（1）：5—9
杨新亭，王林风，王香东.黄原胶与魔芋胶的协效凝胶性研究.食品科学，2001，22（3）：38—40
Noriko Kishida.; Satoshi okjmasn. Preparation of water-soluble methyl konjac glucomannan. Agric Biol Chem, 1978,42(3)：669—670
Kohyama K, In: Gums and Stabilizers of the Food Industry 5, Oxford: IRL Press, 1990
吴贤聪，梁存钧，郭森炎。魔芋葡甘聚糖的提取、鉴定及应用的研究.
李斌，廖勇城，谢笔钧.高效液相色谱法测定魔芋中魔芋葡甘聚糖含量.农牧产品开发.1999,8
12. Vipul Dave,Mihir Sheth,Stephen P. Mccarthy,Jo Ann Ratto and David L. Kapian.Liquid crystalline, rheological and thermal properties of konjac glucomannan . polymer Vol. 39 No.5,pp.1139-11148,1998
13. 沈悦玉, 杨湘庆. 魔芋胶的特征和魔芋凝胶食品. 食品科学, 1995, 16(6): 15- 19.
14. 王照利, 吴万兴, 李科友. 魔芋精粉中甘露聚糖含量测定研究. 食品科学, 1998, 19(3): 56-58.
15. K.Nishinnari ,Yoh Sano ,Kaoru Kohyama. A mixed systerm composed of different molecular weights konjac glucomannan and κ–carrageenan .Ⅱ.Molecular weight dependence of viscoelasticity and thermal properties. Food Hydrocolloids.Vol. 10 no. 2 pp. 229-238,1996
16. K.Nishinnari ,Hiroki Iida ,Kaoru Kohyama. A mixed systerm composed of different molecular weights konjac glucomannan and –carrageenan :large deformation and dynamic viscoelastic study. Food Hydrocolloids.Vol. 7 no. 2 pp. 213-226,1993
庞杰.六偏磷酸钠对魔芋葡甘聚糖干法改性研究.西南农业大学学报，2000年，22（1）：59—61
18. 崔汉钧.魔芋葡甘聚糖固定化环糊精葡基转移酶的研究.天然产物研究与开 发，1993，5（3）：48—54
C.Perols.; B.Piffaut.; J.Scher et al. The potential of enzyme entrapment in konjac cold-melting gel beads. Enzyme and Microbial Technology,1997 20:57—60
周立，蒋磊，郑远旗.魔芋葡苷聚糖凝胶为亲和导析载体与Sepharose 4B的比较研究，Ⅰ.KGM金属螯和层析分离纯化猪血SOD.天然产物研究与开发,1998,10(4):1998,10 (4)：55—58
Wakita M. 公开特许公报（日），昭61—19238（1991,434,354）
Tomoda T. 公开特许公报（日），昭61—21066 （1990,02,212,763）
23 Okaiyi M公开特许公报（日），昭61—19243(1989,434,436)
24. Nakano, Masahiro；Takikawa, Kaori；Arita, Takaichi. Release characteristics of dispersed in konjac gels. Journal of Biomedical Materials Research,1979,13 (5): 811—819
25. Xie, S.S. ; Liu, K.J.; Zhang, Y.X.; Zhang, J.L. et al. Preparation and release test of rotundine-PIC polysaccharide granules. Journal of Macromolecular Science - Pure and Applied Chemistry, 1992,29(10) : 931—938
26. Fanutti C, Ponyi T, Black GW, et al. J Biol Chem, 1995, 270: 29314-29322.
27. 吴襟. β-1,4-D-甘露聚糖酶的纯化和性质[学位论文]. 北京：中科院微生物所, 1997.
AkinoT, Nakamura N, Horikoshi K. β-mannosidase and β-mannanase by an alkalophilic Bacillus sp. Appl.Microbiol.Biotechnol, 1987, 26(4): 323-327.
Firantas S G A, Venzohinskene J I, Paulokonis A B. Prikl.Biokhim.Mikrobiol, 1982, 18: 581-587.
Bruce D, Henk W M, Hilhorst J, et al. Phytochemistry, 1994, 36(4): 829-835.
Barry V, Mccleary. The fine structures of carob and guar galactomannans. Carbohydr Res, 1985, 139: 237-260.
张迎庆，干信，邹群，马丽. 魔芋葡甘低聚糖硫酸酯化衍生物的制备及结构分析. 药物生物技术. 2001,8(4):200-203
M. Yoshimura, K. Nishinari. Dynamic viscoelastic study on the gelation of konjac glucomannan with different molecular weights. Food Hydrocolloids. 13(1999) 227-233
H.Zhang ,M. Yoshimura , K. Nishinari ,M. A. K. Williams ,T. J. Foster , I. T. Norton. Gelation behaviour of konjac glucomannan with different molecular weights.Biopolymers. Vol. 59,38-50(2001)
Mumper R J, Hoffman A S, Puolakkainen P et. al., Calcium-alginate beads for the oral delivery of transforming growth factor-β1: Stabilization of TGF-β1 by the addition of polyacrylic acid within acid-treated beads, J. Control Release, 1994, 30: 241-251
Park K, Robinson J R, Bioadhesive polyners as platforms for oral-controlled drug delivery: Method to study bioadhesion, Int. J. Pharm., 1984,19: 107-127
Chickering D E, Mathiowitz E, Bioadhesive microspheres:ⅠA novel electrobalance-based method to study adhesive interactions between individual microspheres and intestinal mucosa, J. Control. Release, 1995, 34:251-261
Martinsen A, Alginate as immobilization material: Ⅲ. Diffusion properties, Biotechnol. Bioeng., 1992, 39: 186-194
Quong D, Neufeld R J, Skjak-Braek G and Poncelet D, External versus internal source of calcium during the gelation of alginate beads for DNA encapsulation, Biotechnol. Bioeng.,1998, 57: 438-446
Huguet M L, Groboillot A, Poncelet D et. al., Hemoglobin encapsulation in chitosan/calcium alginate beads, J. Appl. Polym. Sci., 1994, 51:1427-1432
Yoshikawa Y, Komuta Y, Nishihara T et. al., Preparation and evaluation of once-a-day injectable microspheres of interferon alpha in rats. J. Drug Target, 1999, 6(6): 449-461
Cho NH, Seong SY, Chun KH et. al., Novel mucosal immunization with polysaccharide-protein conjugates entrapped in alginate microspheres. J. Controlled Release, 1998 ,53(1-3):215-224
Esquisabel A, Hernandez RM, Igartua M et. al., Production of BCG alginate-PLL microcapsules by emulsification/internal gelation, J. Microencapsul, 1997, 14(5): 627-638
Kim HJ, Lee HC, Oh JS et. al., Polyelectroylte complex composed of chitosan and sodium alginate for wound dressing application, J Biomater Sci. Polym. Ed., 1999, 10(5): 543-556
Liu P, Krishnan TR, Alginate-pectin-poly-L-lysine particulate as a potential controlled release formulation, J Pham. Pharmacol., 1999(2): 141-149
Shiraishi S, Imai T, Otagiri M, Controlled-release preparation of indomethacin using calcium alginate gel, Biol. Pharm. Bull, 1993 ,16(11):1164-1168
Takka S, Acarturk F, Calcium alginate microparticles for oral administration: I: Effect of sodium alginate type on drug release and drug entrapment efficiency, J. Microencapsul, 1999, 16(3): 275-290
Tateshita K, Sugawara S, Imai T et. al., Preparation and evaluation of a controlled-release formulation of nifedipine using alginate gel beads, Biol. Pharm. Bull, 1993, 16(4): 420-424
Aslani P, Kennedy RA, Effect of gelation condition and dissolution media on the release of paracetemal from alginate beads, J. Microencapsul, 1996, 13(5):601-614
Sugawara S, Imai T, Otagiri M, The controlled release of prednisolone using alginate gel, Pharm. Res., 1994, 11(2): 272-277
Hari PR, Chandy T, Sharma CP, Chitosan/calcium alginate microcapsules for intestinal delivery of nitrofurantoin, J. Microencapsul, 1996, 13(3):319-329
Pillay V, Fassihi R, In vitro release modulation from crosslinked pellets for site-specific drug delivery to the gastrointestinal tract. I. Comparison of pH-responsive drug release and associated kinetics, J. Controlled Release, 1999, 59(2): 229-242
Sezer AD, Akbuga J, Release characteristics of chitosan treated alginate beads: II. Sustained release of a low molecular drug from chitosan treated alginate beads, J. Microencapsul, 1999 ,16(6): 687-696
Knoll A, Schmidt S, Chapman M et. al., A comparison of two controlled-release delivery systems for the delivery of amiloride to control angiogenesis, Microvasc. Res., 1999 , 58(1):1-9
Fathy M, Safwat SM, el-Shanawany SM et. al., Preparation and evaluation of beads made of different calcium alginate compositions for oral sustained release of tiaramide, Pharm. Dev. Technol., 1998 , 3(3): 355-364
Prisant LM, Bottini B, DiPiro JT et. al., Novel drug-delivery systems for hypertension, Am. J. Med., 1992 , 93(2A): 45S-55S
Kuen Yong L, Won Ho P, Wan Shik H, Polyelectrolyte complexes of sodium alginate with chitosan or its derivatives for microcapasules, J. Appl. Polym. Sci.,
1996, 63: 425-432
Kaneko K, Kanada K, Miyagi M et. al., Formation of water-insoluble gel in dry-coated tablets for the controlled release of theophylline, Chem. Pharm. Bull (Tokyo) , 1998, 46(4): 728-729
Yomota C, Miyazaki T, Okada S, Sustained-release effect of the direct compressed tablet based on chitosan and Na alginate, Yakugaku Zasshi, 1994, 114(4): 257-263
Czarnecki W, Wielgus S, Factors affecting the release of papaverine from spheres, Acta. Pol. Pharm., 1996, 53(5): 331-336
Kishi K, Sonomura T, Nishida N et.al., Alginate gel beads for chemoembolization: initial report, Nippon Igaku Hoshasen Gakkai Zasshi, 1995 , 55(5):300-304
Whitehead L, Fell JT, Collett JH et. al., Floating dosage forms: an in vivo study demonstrating prolonged gastric retention, J. Controlled Release, 1998,55(1):3-12
L Rowsen Moese, Dileep K L, Chandra P Sharma, Bete Cyclodextrin-insulin-encapsulted Chitosan/alginate matrix: for oral delivery of insulin, J. Appl. Polym. Sci., 2000, 75: 1089-1096
曹海辉，王亦农，马建标等，肽类药物口服剂型材料及控制释放性能研究Ⅱ.含有添加剂的壳聚糖-海藻酸盐微囊对胰岛素的控释作用，离子交换与吸附，1999，15（6）：496-503
Gursoy A, Cevik S, Sustained release properties of alginate microspheres and tabletted microspheres of diclofenac sodium, J.Microencapsul, 2000, 17(5): 565-575
Kang Wang and Zhimin He, "Alginate-konjac glucomannan-chitosan beads as controlled release matrix", International Journal of Pharmaceutics, 2002, 244: 117-126
邢琦.尼莫地平缓释微囊的制备及其释放性能的研究. 天津大学硕士论文. 2002