High level expression and facile purification of recombinant silk-elastin-like polymers in auto induction shake flask cultures.
High level expression and facile purification of recombinant silk-elastin-like polymers in auto induction shake flask cultures.
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DOI：
10.1186/2191-0855-3-11
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发表时间：
2013-02-05
期刊：
影响因子：
3.7
通讯作者：
Casal M
中科院分区：
文献类型：
--
作者：
Machado R;Azevedo-Silva J;Correia C;Collins T;Arias FJ;Rodríguez-Cabello JC;Casal M
丝弹性蛋白样聚合物（SELP）是由丝素蛋白（GAGAGS）和哺乳动物弹性蛋白（VPGVG）中发现的重复氨基酸序列基序组成的蛋白质基聚合物。从基础和应用的角度来看，这些聚合物都非常有趣，在生物医学，纳米技术和材料中找到潜在的应用。然而，这种聚合物在这些不同领域中的成功应用需要具有新的增强性能的各种不同形式的现成可用性，并且其可以简单地在工业规模上大量制备。在尝试创建具有改进的性能和适用性的新的聚合物设计中，我们已经开发了四种新的SELP，其中弹性体形成序列聚（VPGVG）被塑料样形成序列聚（VPAVG）替换，并且以不同的比例与丝基序组合。此外，我们优化了简化的生产程序，利用自诱导培养基减少工艺干预，获得的生产水平比以前报道的其他SELP高6倍，体积生产率高于150 mg/L。最后，我们利用这些聚合物已知的增强稳定性，开发了一种简化的非色谱下游处理和纯化方案。一个简单的酸处理允许在一个步骤中细胞破碎和获得相对纯的SELP，随后使用硫酸铵沉淀以提高纯度。这些简化的生产和纯化程序提高了工艺效率，降低了制备这些新型聚合物的成本，并提高了它们的应用潜力。
Silk-elastin-like polymers (SELPs) are protein-based polymers composed of repetitive amino acid sequence motifs found in silk fibroin (GAGAGS) and mammalian elastin (VPGVG). These polymers are of much interest, both from a fundamental and applied point of view, finding potential application in biomedicine, nanotechnology and as materials. The successful employment of such polymers in such diverse fields, however, requires the ready availability of a variety of different forms with novel enhanced properties and which can be simply prepared in large quantities on an industrial scale. In an attempt to create new polymer designs with improved properties and applicability, we have developed four novel SELPs wherein the elastomer forming sequence poly(VPGVG) is replaced with a plastic-like forming sequence, poly(VPAVG), and combined in varying proportions with the silk motif. Furthermore, we optimised a simplified production procedure for these, making use of an autoinduction medium to reduce process intervention and with the production level obtained being 6-fold higher than previously reported for other SELPs, with volumetric productivities above 150 mg/L. Finally, we took advantage of the known enhanced stability of these polymers in developing an abridged, non-chromatographic downstream processing and purification protocol. A simple acid treatment allowed for cell disruption and the obtention of relative pure SELP in one-step, with ammonium sulphate precipitation being subsequently used to enable improved purity. These simplified production and purification procedures improve process efficiency and reduce costs in the preparation of these novel polymers and enhances their potential for application.
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影响因子：
4.9
作者：
Cresce, Arthur von Wald;Dandu, Ramesh;Ghandehari, Hamidreza
通讯作者：
Ghandehari, Hamidreza
DOI：
10.1128/aem.17.2.286-290.1969
发表时间：
1969-01-01
期刊：
APPLIED MICROBIOLOGY
影响因子：
--
作者：
MCDANIEL, LE;BAILEY, EG
通讯作者：
BAILEY, EG
影响因子：
6.2
作者：
McDaniel, Jonathan R.;MacKay, J. Andrew;Quiroz, Felipe Garcia;Chilkoti, Ashutosh
通讯作者：
Chilkoti, Ashutosh
影响因子：
2.9
作者：
Ansaldi, M;Lepelletier, M;Mejean, V
通讯作者：
Mejean, V
影响因子：
1.7
作者：
Machado, R.;Ribeiro, A. J.;Casal, M.
通讯作者：
Casal, M.