利用代谢工程改造贝氏不动杆菌 ADP1 以生产柚皮苷

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2024-10-31 DOI:10.1016/j.mec.2024.e00249
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引用次数: 0

摘要

柚皮苷是一种黄烷酮,也是多种黄酮类化合物的前体,在健康和医药领域具有潜在的应用价值。利用基因工程微生物进行柚皮苷的生物生产被认为是一种前景广阔的策略。涉及查尔酮合成酶(CHS)和查尔酮异构酶(CHI)的柚皮苷合成途径依赖于关键底物丙二酰-CoA 和对香豆酰-CoA 的有效供应。在这项研究中,我们利用了一种土壤细菌--Acinetobacter baylyi ADP1,它所表现出的一些特性使其成为柚皮苷生物合成的合适候选菌株;该菌株天然耐受对香豆酸,并能吸收和代谢对香豆酸,对香豆酸是碱处理木质素中的一种主要化合物,也是生产柚皮苷的前体物质。A. baylyi ADP1 还能产生蜡酯等细胞内脂类,从而为柚皮苷的生物合成提供丙二酰-CoA。此外,该菌株的基因组工程也非常简单。在构建柚皮苷生产菌株的过程中,通过单基因敲除(ΔhcaA)消除了对香豆酸的分解作用,并对植物来源的 CHS 和 CHI 的各种组合进行了评估。来自金丝桃的 CHS 和来自美智子的 CHI 的新型编码基因组合获得了最佳性能,能够在批量培养中利用对香豆酸生产出 17.9 mg/L 的柚皮苷。此外,采用饲料批处理系统后,柚皮苷的产量增加了 3.7 倍(66.4 毫克/升)。这些发现凸显了巴氏杀菌杆菌 ADP1 作为柚皮苷生物合成宿主的潜力,并推动了基于木质素的生物生产。
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Metabolic engineering of Acinetobacter baylyi ADP1 for naringenin production
Naringenin, a flavanone and a precursor for a variety of flavonoids, has potential applications in the health and pharmaceutical sectors. The biological production of naringenin using genetically engineered microbes is considered as a promising strategy. The naringenin synthesis pathway involving chalcone synthase (CHS) and chalcone isomerase (CHI) relies on the efficient supply of key substrates, malonyl-CoA and p-coumaroyl-CoA. In this research, we utilized a soil bacterium, Acinetobacter baylyi ADP1, which exhibits several characteristics that make it a suitable candidate for naringenin biosynthesis; the strain naturally tolerates and can uptake and metabolize p-coumaric acid, a primary compound in alkaline-pretreated lignin and a precursor for naringenin production. A. baylyi ADP1 also produces intracellular lipids, such as wax esters, thereby being able to provide malonyl-CoA for naringenin biosynthesis. Moreover, the genomic engineering of this strain is notably straightforward. In the course of the construction of a naringenin-producing strain, the p-coumarate catabolism was eliminated by a single gene knockout (ΔhcaA) and various combinations of plant-derived CHS and CHI were evaluated. The best performance was obtained by a novel combination of genes encoding for a CHS from Hypericum androsaemum and a CHI from Medicago sativa, that enabled the production of 17.9 mg/L naringenin in batch cultivations from p-coumarate. Furthermore, the implementation of a fed-batch system led to a 3.7-fold increase (66.4 mg/L) in naringenin production. These findings underscore the potential of A. baylyi ADP1 as a host for naringenin biosynthesis as well as advancement of lignin-based bioproduction.
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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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