Understanding carbazole biosynthetic enzymes: potential for a versatile assay of acyl CoAs

了解咔唑生物合成酶：酰基 CoAs 多功能测定的潜力

• 批准号: BB/R00479X/1
• 负责人: Hai Deng
• 金额: $ 49.83万
• 依托单位: University of Aberdeen
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR00479X%2F1
• 关键词: Understanding; carbazole; biosynthetic; enzymes; potential

Fatty acid coenzyme As (acyl-CoAs) ubiquitously exist in living organisms and play central roles in many biochemical reactions. Abnormal accumulation of acyl-CoAs in cells are often the diagnostic symptoms for many diseases (i.e. human metabolic diseases including diabetes, fatty acid oxidation deficiency (FAOD), and cancers). Considerable efforts have gone into developing potential diagnostic tools to measure acyl CoAs so far and yet the outcomes are far from ideal. Acyl CoAs are also the key precursors for microbial fermentation of fatty acid-derived chemicals. Currently many of these chemicals are prepared from fossil-derived feedstocks which require energy intensive processing, and are considered by many as non-sustainable for the future. In this respect, microbial fermentation is an attractive alternative to produce these commodity chemicals and enhanced acyl CoA production is associated with increased production of fatty acid derived chemicals in engineered microbes. In making acyl CoA enhanced, a key challenge is often to quantify the levels of acyl CoA among engineered microbes but researchers have yet to find a reliable way of doing this. In this study, we are going to investigate an enzyme-based system that has the ability to convert acyl-CoAs into coloured aromatic compounds under mild conditions. These aromatic compounds are common motifs in dye materials. Consequently, the incorporation of our enzymatic system into biological samples will offer a versatile and convenient assay to measure acyl CoA concentration within the given samples. The use of bacterial enzymes to accomplish chemical tasks is well established in the food industry. The benefits in terms of sustainable manufacturing are well documented. However, the application of enzymatic processes/industrial biotechnology processes are still underdeveloped in the fine chemical, pharmaceutical, agricultural and bioscience industries. By working out the catalytic capabilities of the two novel enzymes involved in transforming acyl CoAs into coloured aromatic compounds, we will gain the ability to develop their biotechnological potential to be used as a new sensitive assay that will enable us to directly measure acyl CoAs in various biological samples.

脂肪酸辅酶(acyl-COAS)广泛存在于生物体中，在许多生化反应中起着核心作用。细胞中酰基-COA的异常积累通常是许多疾病的诊断症状(即人类代谢性疾病，包括糖尿病、脂肪酸氧化缺乏症(FAOD)和癌症)。到目前为止，在开发潜在的诊断工具来测量酰基COAS方面已经做出了相当大的努力，但结果远不理想。酰基COAs也是微生物发酵脂肪酸衍生化学物质的关键前体。目前，这些化学品中的许多都是从化石衍生的原料中制备的，需要能源密集型加工，许多人认为这些原料在未来是不可持续的。在这方面，微生物发酵是生产这些商品化学品的一个有吸引力的替代方案，而酰化辅酶A产量的提高与工程微生物中脂肪酸衍生化学品产量的增加有关。在增强酰基辅酶A的过程中，一个关键的挑战往往是量化工程微生物中的酰基辅酶A水平，但研究人员尚未找到一种可靠的方法来做到这一点。在这项研究中，我们将研究一种基于酶的体系，该体系能够在温和的条件下将酰基COA转化为有色的芳香族化合物。这些芳香族化合物是染料材料中常见的基元。因此，将我们的酶系统整合到生物样品中将提供一种通用和方便的方法来测量给定样品中的酰基辅酶A浓度。使用细菌酶来完成化学任务在食品工业中已经得到了很好的确立。在可持续制造方面的好处是有据可查的。然而，在精细化工、制药、农业和生物科学行业，酶法/工业生物技术的应用仍然不发达。通过研究这两种新型酶的催化能力，我们将能够开发它们的生物技术潜力，作为一种新的灵敏的分析方法，使我们能够直接测定各种生物样品中的酰基COAs。

项目成果

Characterization of a class II ketol-acid reductoisomerase from Mycobacterium tuberculosis.

• DOI: 10.1039/d1ra08876a
• 发表时间: 2022-03-31
• 期刊: RSC advances
• 影响因子: 3.9
• 作者: Valera A;Wang S;Carr R;Trembleau L;Deng H
• 通讯作者: Deng H

Chemo-Enzymatic Synthesis of Bioactive Carbazole Derivatives

• DOI: 10.3390/synbio2010002
• 发表时间: 2024-01
• 期刊: SynBio
• 作者: Saad Alrashdi;Federica Casolari;K. Kyeremeh;Hai Deng

Aminoacyl chain translocation catalysed by a type II thioesterase domain in an unusual non-ribosomal peptide synthetase.

一种不寻常的非核糖体肽合成酶中 II 型硫酯酶结构域催化的氨酰基链易位

• DOI: 10.1038/s41467-021-27512-0
• 发表时间: 2022-01-10
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Wang S;Brittain WDG;Zhang Q;Lu Z;Tong MH;Wu K;Kyeremeh K;Jenner M;Yu Y;Cobb SL;Deng H
• 通讯作者: Deng H