Biosynthesis of Polyhydroxyalkanoate Terpolymer from Methanol via the Reverse β-Oxidation Pathway in the Presence of Lanthanide.
Biosynthesis of Polyhydroxyalkanoate Terpolymer from Methanol via the Reverse β-Oxidation Pathway in the Presence of Lanthanide.
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在镧系元素存在下通过反向 β-氧化途径从甲醇生物合成聚羟基脂肪酸酯三元共聚物。
DOI：
10.3390/microorganisms10010184
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发表时间：
2022-01-15
期刊：
影响因子：
4.5
通讯作者：
Fukui T
中科院分区：
文献类型：
--
作者：
Orita I;Unno G;Kato R;Fukui T
Methylorubrum extorquens AM1是从甲醇生产增值产品的有吸引力的平台。我们以前证明了M。装配有具有广泛底物特异性的PHA合酶的扭脱醌在甲醇上合成由（R）-3-羟基丁酸酯和小部分（R）-3-羟基戊酸酯（3HV）和（R）-3-羟基己酸酯（3HHx）单元组成的聚羟基链烷酸酯（PHA）。这项研究进一步改造了M。extorquens的生物合成的PHA与更高的3HV和3HHx组成集中在EMC途径参与C1同化。引入乙基丙二酰辅酶A脱羧酶，催化EMC途径中的逆向反应，旨在增加细胞内丙酰/丁酰辅酶A前体，不影响PHA组合物。然后通过异源表达源自真养罗尔斯通氏菌的四个基因来增强2-烯酰-CoA的反向β-氧化途径和随后的（R）-特异性水合，以将丙酰/丁酰-CoA转化为相应的（R）-3-羟酰-CoA单体。所得菌株产生具有较高的3HV和3HHx组成的PHA，而甲基营养生长严重受损。有趣的是，这种生长障碍通过添加La 3+而恢复，而对PHA生物合成没有负面影响，这表明La 3+激活了EMC途径。工程M。以甲醇为唯一碳源，在La~（3+）存在下，扭脱醌合成了3HV含量为5.4mol%，3HHx含量为0.9%，3HHx含量为41%的PHA.
Methylorubrum extorquens AM1 is the attractive platform for the production of value-added products from methanol. We previously demonstrated that M. extorquens equipped with PHA synthase with broad substrate specificity synthesized polyhydroxyalkanoates (PHAs) composed of (R)-3-hydroxybutyrate and small fraction of (R)-3-hydroxyvalerate (3HV) and (R)-3-hydroxyhexanoate (3HHx) units on methanol. This study further engineered M. extorquens for biosynthesis of PHAs with higher 3HV and 3HHx composition focusing on the EMC pathway involved in C1 assimilation. The introduction of ethylmalonyl-CoA decarboxylase, catalyzing a backward reaction in the EMC pathway, aiming to increase intracellular propionyl/butyryl-CoA precursors did not affect PHA composition. Reverse β-oxidation pathway and subsequent (R)-specific hydration of 2-enoyl-CoA were then enhanced by heterologous expression of four genes derived from Ralstonia eutropha for the conversion of propionyl/butyryl-CoAs to the corresponding (R)-3-hydroxyacyl-CoA monomers. The resulting strains produced PHAs with higher 3HV and 3HHx compositions, while the methylotrophic growth was severely impaired. This growth impairment was interestingly restored by the addition of La3+ without a negative impact on PHA biosynthesis, suggesting the activation of the EMC pathway by La3+. The engineered M. extorquens synthesized PHA terpolymer composed of 5.4 mol% 3HV and 0.9% of 3HHx with 41% content from methanol as a sole carbon source in the presence of La3+.
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影响因子：
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