SU Jun-cai,DONG Chao,SHI Yan-mao,et al.Extraction of Nattokinase by Soybean Protein Cross-linked Chitosan Microsphere[J].Soybean Science,2011,30(04):652-656,662.[doi:10.11861/j.issn.1000-9841.2011.04.0652]
大豆蛋白偶联壳聚糖微球介质提取纳豆激酶的研究
- Title:
- Extraction of Nattokinase by Soybean Protein Cross-linked Chitosan Microsphere
- 文章编号:
- 1000-9841(2011)04-0652-05
- 分类号:
- R945
- 文献标志码:
- A
- 摘要:
- 采用液体反相悬浮法制取壳聚糖微球,在微球表面偶联纳豆激酶的亲和配体—大豆蛋白,作为纳豆溶栓酶的亲和吸附介质。结果表明:壳聚糖微球本身对纳豆激酶没有特异吸附,偶联大豆蛋白的壳聚糖微球对发酵粗酶液中的纳豆激酶的吸附性能符合Langmuir方程,吸附平衡时间约为60 min,最大吸附量为3 926.56 U?g-1。纳豆激酶发酵液的上样量与柱体积比为1∶9时,纳豆激酶的收率可以达到52.3%,纯化倍数可以达到18.1倍。
- Abstract:
- Chitosan microspheres were obtained by inverse phase suspension method.After being cross-linked with soybean protein on its surface,the chitosan microspheres became affinity adsorbent of Nattokinase.This study verified that there were no affinity binding between the chitosan microspheres and Nattokinase.The affinity adsorption was researched between the soybean protein cross-linked microspheres and Nattokinase.Results indicated that the adsorption isotherm was accorded with Langmuir,the time of adsorption balance was 60 min,the maximum of adsorption capacity was 3926.56 U?g-1.When the ratio of sample weight and column volume was 1∶9,the purification fold and recovery of Nattokinase activity reached 18.1 times and 52.3%,respectively.
参考文献/References:
[1]Fujita M,Nomura K,Kong K.Purification and characterization of a strong fibrinolytic enzyme(Nattokinase)in the vegetable cheese natto,a popular soybean fermented food in Japan[J].Biochemical and Biophysical Research Communications,1993,197(3):1340-1347. [2]Sumi H,Hamada H,Tsuhima H,et al.A novel fibrinolytic enzyme(Nattokinase)in the vegetable cheese Natto:a typical and popular soybean food in the Japanese diet[J].Experientia,1987,43(20):1110-1111. [3]高大海,梅乐和,盛清,等.硫酸铵沉淀和层析法分离纯化纳豆激酶的研究[J].高校化学工程学报,2006,20(1):63-67.(Gao D H,Mei Y H,Sheng Q,et al.Separation and purification of Nattokinase produced by bacillus subtilis with ammonium sulfate precipitation and chromatography[J].Journal of Chemical Engineering of Chinese Universities,2006,20(1):63-67.) [4]阳承利,邢建民,刘俊果,等.纳豆激酶分离纯化技术的研究进展[J].现代化工,2004,24(2):23-25.(Yang C L,Xing J M,Liu J G,et al.Progress in separation and purification technology of nattokinase[J].Modern Chemical Industry,2004,24(2):23-25.) [5]张锋,金杰,解成俊.纳豆激酶的最新研究进展[J].中国调味品,2008,347:29-34.(Zhang F,Jin J,Xie C J.New research progress in nattokinase[J].China Condiment,2008,347:29-34.) [6]孙彦.生物分离工程[M],北京:化学工业出版社,2005:189-191.(Sun Y.Bioseparation engineering[M].Beijing:Chemical Industry Press,2005:189-191.) [7]陈勇,徐燚,应汉杰,等.亲和层析研究进展[J].离子交换与吸附,2001,17(3):276-280.(Chen Y,Xu Y,Ying H J,et al.Researches and developments of affinity chromatography[J].Ion Exchange and Adsorption,2001,17(3):276-280.) [8]陆健.蛋白质纯化技术及应用[M].北京:化学工业出版社,2005:120-122.(Lu J.Protein purification and application[M].Beijing:Chemical Industry Press,2005:120-122.) [9]马跃华,董超,杨明,等.纳豆激酶的亲和吸附工艺研究[J].大豆科学,2010,29(2):306-309.(Ma Y H,Dong C,Yang M,et al.Adsorption of Nattokinase by soybean microspheres[J].Soybean Science,2010,29(2):306-309.) [10]唐雪娇,曹梦,毕成良,等.球形交联壳聚糖吸附剂的多胺化及其对Ni2+的吸附性能[J].离子交换与吸附,2008,24(1):47-54.(Tang X J,Cao M,Bi C L,et al.Study on the synthesis and characterization of a novel polyaminated and Ni2+templated cross-linked chitosan bead and its adsorption[J].Ion Exchange and Adsorption,2008,24(1):47-54.) [11]党明岩,张延安,李薇,等.新型交联壳聚糖树脂颗粒对铂的吸附行为[J].过程工程学报,2008,8(1):60-64.(Dang M Y,Zhang Y A,Li W,et al.Adsorption of platinum by novel cross-linked chitosan resin particles[J].The Chinese Journal of Process Engineering,2008,8(1):60-64.) [12]刘健敏,钟芳,麻建国.大豆生理活性肽的研究Ⅱ抗氧化性和ACE抑制活性的初步研究[J].无锡轻工大学学报,2004,23(4):50-55.(Liu J M,Zhong F,Ma J G,et al.Soybean bioactive peptides(Ⅱ)-antioxidant activity and ACE inhibitory activity of the hydrolysates[J].Journal of Wuxi University of Light Industry,2004,23(4):50-55.) [13]杨明,董超,史延茂,等.纤维蛋白平板法测定纳豆激酶方法的改进[J].中国酿造,2008,(7):77-80.(Yang M,Dong C,Shi Y M,et al.Improvement on the method of measuring the activity of Nattokinase with agarose-fibrin plate[J].China Brewing,2008,(7):77-80.) [14]吕莹,张露,冯雷,等.纳豆激酶的纯化及性质研究[J].食品与发酵工业,2004,30(3):122-124.(Lu Y,Zhang L,Feng L,et al.Purification and characterization of Nottokinase from Bacillus subtilis[J].Food and Fementation Industries,2004,30(3):122-124.) [15]华南理工大学.一种利用亲和层析法分离纯化纤溶酶的方法[P].中国发明专利申请公开说明书,CN101067131A.2007-11-7.(South China University of Technology.Purification of fibrinolytic enzyme by affinity chromatography[P].China Patent,CN101067131A.2007-11-7.) [16]上海交通大学.降纤酶的亲和分离材料和降纤酶的亲和纯化方法[P].中国发明专利申请公开说明书,CN1884507A.2006-12-27.(Shanghai Jiao Tong University.Media and purification of defibrase with affinity chromatography[P].China Patent,CN1884507A.2006-12-27.)
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备注/Memo
基金项目:河北省科学技术研究与发展计划资助项目(08276413D)。