由铁胺合成酶的铁酶域催化的酸环化
J W Trauger1, R M Kohli, H D Mootz
1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
这项研究表明,切除的铁酶 (TE) 域可以独立催化-铁的宏循环. 这一发现对于理解TE基质特异性和设计新的天然产品生物合成至关重要.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 自然产品生物合成 自然产品生物合成
背景情况:
- 多域大合成酶合成宏循环的自然产物,其中有碳氧终端化酶 (TE) 域介导循环和释放.
- TE域的独立催化能力和基质特异性在很大程度上仍未确定,这是由于它们在更大的酶复合体中集成.
研究的目的:
- 调查切除的硫酶 (TE) 域是否可以独立于上游酶域来催化宏循环.
- 为了确定来自铁胺合成酶的TE域的基质特异性,以在工程生物合成中潜在的应用.
主要方法:
- 从铁胺合成酶中切除TE域.
- 评估孤立的TE域在循环分解的去酸铁和二聚化/循环分解的五酸铁中的催化活性.
- 系统地改变-基质以分析循环化速率并识别关键残留物.
主要成果:
- 切除的TE域有效地催化了十甲-三的循环生成,形成铁氨酸A.
- TE域还催化了五-硫的二元化和随后的循环化,以形成格拉米西丁S.
- 分析显示,在基底中,只有两个特定的残留物对有效的循环形成至关重要.
结论:
- 切除的TE域具有对宏循环的内在催化活性,独立于上游的酶机制.
- 鉴定到的基质特异性概况表明,TE域对于在工程生物合成中循环利用多种基质具有广泛的适用性.
- 这项工作为通过合成生物学方法设计新型循环和天然产品提供了基础.
更多相关视频
08:02Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory
Published on: August 23, 2018
10:21Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
Published on: June 20, 2019
相关概念视频
Introduction to Mechanisms of Enzyme Catalysis
Acid Halides to Carboxylic Acids: Hydrolysis
As shown below, the mechanism involves a nucleophilic attack by water at the carbonyl carbon to form a tetrahedral intermediate. This is followed by the reformation of the carbon–oxygen π bond along with the departure of a halide ion. A final proton transfer step yields...
Acid Halides to Esters: Alcoholysis
Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...
Phase II Reactions: Sulfation and Conjugation with α-Amino Acids
Acid Mine Drainage
![Protein Film Infrared Electrochemistry Demonstrated for Study of H2 Oxidation by a [NiFe] Hydrogenase](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F55858.jpg&w=3840&q=50)