大腹园蛛梨状腺丝重组蛋白的表达纯化与纺丝  被引量:1

Expression,purification and spinning of recombinant pyriform spidroins from Araneus ventricosus

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作  者:易拓 朱红年 孟清[1] YI Tuo;ZHU Hongnian;MENG Qing(Department of Bioengineering, College of Chemistry Chemical Engineering and Bioengineering, Donghua University, Shanghai, 201620)

机构地区:[1]东华大学化学化工与生物工程学院,上海201620

出  处:《工业微生物》2020年第5期27-35,共9页Industrial Microbiology

摘  要:圆网蜘蛛能分泌生产六种不同的蛛丝纤维和一种粘胶,其中梨状腺丝参与附着盘的形成,蜘蛛利用附着盘将蜘蛛丝固定于物体表面。迄今为止,相对于广泛研究的主壶腹腺丝,鲜有关于重组梨状腺丝蛋白与重组梨状腺丝力学性能的研究报道。本研究以大腹园蛛梨状腺丝蛋白(PySp1)一个重复单元的编码DNA为模板,通过无缝克隆技术构建能够表达含有2—3个相同重复单元的梨状腺丝重组蛋白(2R、3R)的表达质粒,并在大肠杆菌工程菌中表达。经纯化获得较高纯度的重组蛋白2R和3R。使用湿法纺丝制成丝纤维,对制得的纤维以力学性能测试、FTIR检测以及热重(TGA)分析等进行表征。结果表明,重组丝纤维表现出优良的力学性能,并具有和天然蛛丝纤维相似的二级结构和一般聚合物类似的热稳定性。该研究为人工合成生物纤维提供了理论依据。An orb-weaving spider can secrete and produce six different kinds of silks and a sort of sticky glue.Among them,pyriform silk is involved in the formation of the attachment disc and the spider uses the attachment disc to fix the spider silk on the surface of the object.So far,compared with the widely studied MaSp silk,there are few reports on mechanical properties of recombinant pyriform spidroin and silk.Based on the fragment of DNA of one repeat unit of PySp1 from A.ventricosus,plasmids which could be used to produce recombinant pyriform spidroins,were constructed through seamless clone.The recombinant spidroins composed of 2-3 identical repeat units were produced in engineered E.coli strain cells and named as 2R and 3R.Purified spidroins with high purity were spun into silk-like fibers with a wet-spinning process.Tensile property test,fourier-transform infrared spectroscopy and thermal gravity analysis were used to characterize the fibers.Experimental results indicated that recombinant fibers exhibited excellent mechanical properties;the content of secondary structures existing in the fibers resembled that of nature spider silk,and thermal stability of both fibers was similar to that of general polymers.This study would provide theoretical foundation to synthetic biological fibers in the future.

关 键 词:梨状腺丝重组蛋白(2R、3R) 重组梨状腺丝 力学性能 

分 类 号:TQ3[化学工程]

 

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