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Expanding the toolbox for making pol...

Rude, Mathew Allen.

Expanding the toolbox for making polyketides in Escherichia coli: Heterologous expression of ansamycin polyketides and engineered synthesis of 2-substituted malonyl-CoAs.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Expanding the toolbox for making polyketides in Escherichia coli: Heterologous expression of ansamycin polyketides and engineered synthesis of 2-substituted malonyl-CoAs.
作者:	Rude, Mathew Allen.
面頁冊數:	129 p.
附註:	Adviser: Chaitan Khosla.
附註:	Source: Dissertation Abstracts International, Volume: 67-09, Section: B, page: 5251.
Contained By:	Dissertation Abstracts International67-09B.
標題:	Chemistry, Biochemistry.
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3235333
ISBN:	9780542895708

Expanding the toolbox for making polyketides in Escherichia coli: Heterologous expression of ansamycin polyketides and engineered synthesis of 2-substituted malonyl-CoAs.
Rude, Mathew Allen.

Expanding the toolbox for making polyketides in Escherichia coli: Heterologous expression of ansamycin polyketides and engineered synthesis of 2-substituted malonyl-CoAs. - 129 p.

Adviser: Chaitan Khosla.

Thesis (Ph.D.)--Stanford University, 2006.

Escherichia coli has emerged a powerful heterologous host for the production of polyketides. Driving the interest in using E. coli to make polyketides is our unparalleled understanding of E. coli's biology and the existence of numerous tools to manipulate both its genetics and its metabolism. To-date, only relatively simple polyketide products, like 6-DEB, have been made in E. coli. In this study, tools were developed to produce more complicated polyketides. This allowed the synthesis of ansamycin polyketides and 6-DEB derivatives.

ISBN: 9780542895708Subjects--Topical Terms:

226900
Chemistry, Biochemistry.

Expanding the toolbox for making polyketides in Escherichia coli: Heterologous expression of ansamycin polyketides and engineered synthesis of 2-substituted malonyl-CoAs.
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245 1 0 $a Expanding the toolbox for making polyketides in Escherichia coli: Heterologous expression of ansamycin polyketides and engineered synthesis of 2-substituted malonyl-CoAs.
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520 # $a Escherichia coli has emerged a powerful heterologous host for the production of polyketides. Driving the interest in using E. coli to make polyketides is our unparalleled understanding of E. coli's biology and the existence of numerous tools to manipulate both its genetics and its metabolism. To-date, only relatively simple polyketide products, like 6-DEB, have been made in E. coli. In this study, tools were developed to produce more complicated polyketides. This allowed the synthesis of ansamycin polyketides and 6-DEB derivatives.
520 # $a Many polyketide pathways require 2-substituted malonyl-CoAs substrates other then methylmalonyl-CoA. A system was created to generate 2-substituted malonyl-CoAs intracellularly in E. coli. In this system, 2-substituted malonic acid compounds are added to the media, imported into the cell, and then ligated to CoA using a non-specific malonyl-CoA ligase from Bradyrhizobium japonicum. To test the ability of this system to generate ethylmalonyl-CoA, module 6 of the DEBS enzymes, which produced 6-DEB, was engineered to accept ethylmalonyl-CoA to make 6-ethyl-DEB.
520 # $a Polyketide natural products play an important role in the treatment of a wide range of human physiological disorders. They are widely used as antimicrobial, antifungal, immunosuppressants, and anti-cancer agents, as well as cholesterol lowering agents. Polyketides also play an important role in animal health and agriculture. Current organic synthesis technologies are unable to generate commercially viable quantities of most polyketide compounds. As a result, the natural sources of these compounds, usually bacteria and fungi, are employed as biocatalysts for large-scale production. However, in the past two decades an explosive growth in the cloning and sequencing of genes responsible for polyketide production, and our increasing knowledge of the catalytic mechanisms governing the biosynthesis of polyketides, has made heterologous expression of polyketide genes a reality.
520 # $a The synthesis of ansamycin polyketides in E. coli presents new challenges. Strategies for expressing proteins greater then 500 kDa were developed and the polyketide substrates 3-amino-5-hydroxybenzoic acid and methoxymalonyl-ACP were engineered into E. coli. A rifamycin intermediate P8/1-OG was generated in E. coli using these tools and a preliminary system for making pro-geldanamycin was assembled.
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