Optimization of Purification Technology of Lobster Sauce Streptokinase by Macroporous Resin

WANG Peng-jiao; MENG Xiao-xia; ZHANG Min; JIANG Jian-min; GAO Xiu-li

doi:10.13422/j.cnki.syfjx.2014110030

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Optimization of Purification Technology of Lobster Sauce Streptokinase by Macroporous Resin

更新时间：2024-01-04

- Optimization of Purification Technology of Lobster Sauce Streptokinase by Macroporous Resin
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 20, Issue 11, Pages: 30-33(2014)
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- DOI：10.13422/j.cnki.syfjx.2014110030
  CLC： R284;TQ461
- Published：2014
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WANG Peng-jiao, MENG Xiao-xia, ZHANG Min, et al. Optimization of Purification Technology of Lobster Sauce Streptokinase by Macroporous Resin[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(11): 30-33.
DOI：

WANG Peng-jiao, MENG Xiao-xia, ZHANG Min, et al. Optimization of Purification Technology of Lobster Sauce Streptokinase by Macroporous Resin[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(11): 30-33. DOI： 10.13422/j.cnki.syfjx.2014110030.

摘要

目的：优选豆豉溶栓酶的大孔树脂纯化工艺。方法：以湿比吸附量为指标筛选大孔树脂型号；以豆豉溶栓酶比活性为指标，采用单因素试验考察吸附性能（上样体积、吸附时间）和洗脱性能（洗脱剂种类、洗脱剂pH、洗脱速度、洗脱剂体积），利用SDS-PAGE电泳对纯化效果进行分析。结果：HPD-400型大孔树脂对豆豉溶栓酶具有很好的吸附作用，优选的工艺条件为上样量2 BV，吸附时间2 h，加水2 BV洗去杂质，加pH 8.0的Tris-HCl-30%乙醇缓冲液4 BV以2 BV·h-1流速洗脱；豆豉溶栓酶比活性2 254.29 IU·mg-1，纯化数44.17倍，发酵液20 mL平均可得到蛋白质2.86 mg，电泳检测显示豆豉溶栓酶呈现单一条带，相对分子质量28 kD。结论：HPD-400型大孔树脂可用于豆豉溶栓酶的分离纯化，优选的工艺条件稳定可行。

Abstract

Objective:To optimize purification technology of lobster sauce streptokinase by macroporous resin. Method:Type of macroporous resin was screened by taking moisture adsorption as index;With specific activity of lobster sauce streptokinase as index

single factor tests were adopted to optimize purification process parameters

such as adsorption performance(loading quantity of sample

adsorption time) and elution performance(species

pH and volume of eluent

elution rate)

purification effect was analyzed by SDS-PAGE electrophoresis. Result:HPD-400 macroporous resin had a good adsorbability to lobster sauce streptokinase

optimum purification conditions were as follows:sample volume of 2 BV

adsorption time of 2 h

removed impurities with 2 BV of water

eluted with 4 BV of Tris HCl-30% ethanol buffer(pH 8.0) with adsorption speed of 2 BV·h-1;under these conditions

specific activity of lobster sauce streptokinase was 2 254.29 U·mg-1

purification multiple was 44.17 times

we could receive 2.86 mg of protein in 20 mL of fermentation liquor

lobster sauce streptokinase showed a single strip in electrophoretogram whose relative molecular weight was 28 kD. Conclusion:HPD-400 macroporous resin could be adopted to purify lobster sauce streptokinase

this optimized purification technology was stable and feasible.

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Related Institution

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School of Pharmacy， Jiangxi University of Chinese Medicine

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