检索结果-南通市图书馆

Improving the reaction mix of a Pichia pastoris cell-free system using a design of experiments approach to minimise experimental effort

引用

Synthetic and Systems Biotechnology 2020年第3期5卷 137-144页

作者： Alex J.Spice Rochelle Aw daniel g.bracewell Karen M.Polizzi Department of Chemical Engineering Imperial College LondonLondonUK Imperial College Centre for Synthetic Biology Imperial College LondonUK Department of Biochemical Engineering University College LondonLondonUK

A renaissance in cell-free protein synthesis(CFPS)is underway,enabled by the acceleration and adoption of synthetic biology methods.CFPS has emerged as a powerful platform technology for synthetic gene network design,biosensing and on-demand biomanufacturing.Whilst primarily of bacterial origin,cell-free extracts derived from a variety of host organisms have been explored,aiming to capitalise on cellular diversity and the advantageous properties associated with those organisms.However,cell-free extracts produced from eukaryotes are often overlooked due to their relatively low yields,despite the potential for improved protein folding and posttranslational modifications.Here we describe further development of a Pichia pastoris cell-free platform,a widely used expression host in both academia and the biopharmaceutical industry.Using a minimised Design of Experiments(DOE)approach,we were able to increase the productivity of the system by improving the composition of the complex reaction mixture.This was achieved in a minimal number of experimental runs,within the constraints of the design and without the need for liquid-handling robots.In doing so,we were able to estimate the main effects impacting productivity in the system and increased the protein synthesis of firefly luciferase and the biopharmaceutical HSA by 4.8-fold and 3.5-fold,respectively.This study highlights the P.pastoris-based cell-free system as a highly productive eukaryotic platform and displays the value of minimised DOE designs.

关键词： Cell-free protein synthesis Pichia pastoris Synthetic biology Design of experiments(DOE)

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：