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植物根瘤菌属 Z69 的 prpC 突变株的丙酸代谢和聚 3-羟基丁酸-co-3-羟基戊酸的生产。

Propionic acid metabolism and poly-3-hydroxybutyrate-co-3-hydroxyvalerate production by a prpC mutant of Herbaspirillum seropedicae Z69.

机构信息

Unidad Microbiología Molecular - Depto. BIOGEM, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Av. Italia 3318, Montevideo, CP11600, Uruguay.

Unidad Microbiología Molecular - Depto. BIOGEM, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Av. Italia 3318, Montevideo, CP11600, Uruguay.

出版信息

J Biotechnol. 2018 Nov 20;286:36-44. doi: 10.1016/j.jbiotec.2018.09.008. Epub 2018 Sep 18.

Abstract

Polyhydroxyalkanoates (PHAs) are thermoplastic polyesters produced by a wide range of bacteria as carbon and energy reserves. PHA accumulation is typically increased under unbalanced growth conditions and with carbon source in excess. Although polyhydroxybutyrate (PHB) could be used for specific applications, it is brittle and not a useful alternative for plastics like polypropylene. Far more useful polypropylene-like PHAs, are copolymers composed of 3-hydroxybutyrate and 3-hydroxyvalerate, P(3HB-co-3HV). Propionic acid is one of the carbon sources that can be used to generate 3HV. A mutant derived from Herbaspirillum seropedicae Z69, a strain previously described as capable of producing P(3HB-co-3HV) from propionic acid, was constructed to increase 3HV biosynthetic efficiency. The strategy involved elimination of a catabolic route for propionyl-CoA by deficiency marker exchange of a selected gene. The mutant (Z69Prp) was constructed by elimination of the 2-methylcitrate synthase (PrpC) gene of the 2-methylcitrate cycle for propionate catabolism. Strain Z69Prp was unable to grow on sodium propionate, but in cultures with glucose-propionate accumulated 50% of its dry weight as copolymer. Z69Prp had 14.1 mol% 3HV; greater than that of strain Z69 (2.89 mol%). The 3HV yield from propionic acid (Y) was 0.80 g g, and below the maximum theoretical value (1.35 g g).

摘要

聚羟基脂肪酸酯(PHAs)是由多种细菌作为碳源和能量储备合成的热塑性聚酯。在不平衡生长条件下和碳源过剩时,PHA 的积累通常会增加。虽然聚羟基丁酸酯(PHB)可用于特定应用,但它易碎,不能替代聚丙烯等塑料。更有用的类似聚丙烯的 PHAs 是由 3-羟基丁酸酯和 3-羟基戊酸酯组成的共聚物,P(3HB-co-3HV)。丙酸是可用于生成 3HV 的碳源之一。从 Herbaspirillum seropedicae Z69 衍生出一种突变体,该菌株以前被描述为能够从丙酸生产 P(3HB-co-3HV),构建该突变体是为了提高 3HV 生物合成效率。该策略涉及通过选择基因的缺陷标记交换消除丙酰辅酶 A 的分解代谢途径。通过消除用于丙酸分解代谢的 2-甲基柠檬酸合酶(PrpC)基因,构建了突变体(Z69Prp)。菌株 Z69Prp 不能在丙酸钠上生长,但在含有葡萄糖-丙酸的培养物中,其干重的 50%积累为共聚物。Z69Prp 有 14.1 mol%的 3HV;高于 Z69 菌株(2.89 mol%)。从丙酸获得的 3HV 产率(Y)为 0.80 g g,低于最大理论值(1.35 g g)。

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