Exploiting the sequence of naturally occurring elastin: Construction, production and characterization of a recombinant thermoplastic protein-based polymer
Exploiting the sequence of naturally occurring elastin: Construction, production and characterization of a recombinant thermoplastic protein-based polymer
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DOI：
10.4028/www.scientific.net/jnanor.6.133
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发表时间：
2009-01-01
影响因子：
1.7
通讯作者：
Casal, M.
中科院分区：
文献类型：
--
作者：
Machado, R.;Ribeiro, A. J.;Casal, M.
基因工程用于生产具有精确氨基酸组成、序列和长度的弹性蛋白样聚合物（ELP），从而导致绝对控制MW和立体化学。编码（VPAVG）（20）的合成单体DNA序列用于构建对序列和大小具有精确控制的多联体基因文库。基于ELP的可逆逆温度转变特性，在大肠杆菌中生物产生具有220个重复的VPAVG的较高分子量聚合物，并通过热离心和冷离心循环纯化。使用中心复合设计方法探索了用于细菌细胞培养基的低成本碳源如乳糖和甘油的使用，从而允许优化发酵条件。由于其自组装行为接近33摄氏度的稳定的球形微粒的大小类似于1 μ m的获得，再溶解时，一个强大的过冷实现。所产生的聚合物显示出滞后行为，热吸收/释放组分取决于聚合物溶液的盐浓度。
Genetic engineering was used to produce an elastin-like polymer (ELP) with precise amino acid composition, sequence and length, resulting in the absolute control of MW and stereochemistry. A synthetic monomer DNA sequence encoding for (VPAVG)(20), was used to build a library of concatemer genes with precise control on sequence and size. The higher molecular weight polymer with 220 repeats of VPAVG was biologically produced in Escherichia coli and purified by hot and cold centrifugation cycles, based on the reversible inverse temperature transition property of ELPs. The use of low cost carbon sources like lactose and glycerol for bacteria cells culture media was explored using Central Composite Design approach allowing optimization of fermentation conditions. Due to its self-assembling behaviour near 33 degrees C stable spherical microparticles with a size similar to 1 mu m were obtained, redissolving when a strong undercooling is achieved. The polymer produced showed hysteresis behaviour with thermal absorbing/releasing components depending on the salt concentration of the polymer solution.