Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21.
Polyhydroxybutyrate production by recombinant Escherichia coli based on genes related to synthesis pathway of PHB from Massilia sp. UMI-21.
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DOI：
10.1186/s12934-023-02142-x
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发表时间：
2023-07-14
影响因子：
6.4
通讯作者：
中科院分区：
文献类型：
--
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聚羟基丁酸酯（Polyhydroxybutyrate，简称PHB）是目前最常见的由天然细菌生产的聚合物，由于其相似的材料性能和生物降解性，可替代传统的石化基塑料。一株新发现的细菌Massilia sp. UMI-21能利用淀粉、麦芽三糖、麦芽糖等物质生产聚羟基丁酸酯，有望成为降解海藻的生物塑料生产菌。然而，在Massilia sp. UMI-21中参与PHB代谢的基因仍然不清楚。在本研究中，我们组装和注释了Massilia sp. UMI-21的基因组，鉴定了与PHB代谢相关的基因，并成功构建了含有Massilia sp. UMI-21的PHB相关基因（phaA 2，phaB 1和phaC 1）的重组大肠杆菌，其产物产量增加了139.41%。此外，将vgb基因（编码Vitrechina血红蛋白）导入基因工程E。coli的菌体干重提高了117.42%，类聚β-羟基丁酸（PHB-like）产量提高了213.30%，产物含量提高了44.09%。重组E. pETDuet 1-phaA 2 phaB 1-phaC 1和pETDuet 1-phaA 2 phaB 1-phaC 1-vgb经傅立叶变换红外光谱和质子核磁共振谱分析鉴定为聚羟基丁酸酯。此外，还测定了从Massilia sp. UMI-21、重组E.大肠杆菌DH 5 α-pETDuet 1-phaA 2 phaB 1-phaC 1和DH 5 α-pETDuet 1-phaA 2 phaB 1-phaC 1-vgb的热稳定性分别为276.5、278.7和286.3 °C，具有良好的热稳定性。本研究以产聚羟基丁酸酯（PHB）的Massilia sp. UMI-21为研究对象，利用其基因组信息，构建了一株新的重组菌。这种基因工程E. coli菌株可作为大肠杆菌的有效候选菌株。大肠杆菌细胞工厂生产的PHB的快速细胞生长和高产量的PHB。在线版本包含补充材料，可通过10.1186/s12934-023-02142-x获得。
Polyhydroxybutyrate (PHB) is currently the most common polymer produced by natural bacteria and alternative to conventional petrochemical-based plastics due to its similar material properties and biodegradability. Massilia sp. UMI-21, a newly found bacterium, could produce PHB from starch, maltotriose, or maltose, etc. and could serve as a candidate for seaweed-degrading bioplastic producers. However, the genes involved in PHB metabolism in Massilia sp. UMI-21 are still unclear. In the present study, we assembled and annotated the genome of Massilia sp. UMI-21, identified genes related to the metabolism of PHB, and successfully constructed recombinant Escherichia coli harboring PHB-related genes (phaA2, phaB1 and phaC1) of Massilia sp. UMI-21, which showed up to 139.41% more product. Also, the vgb gene (encoding Vitreoscilla hemoglobin) was introduced into the genetically engineered E. coli and gained up to 117.42% more cell dry weight, 213.30% more PHB-like production and 44.09% more product content. Fermentation products extracted from recombinant E. coli harboring pETDuet1-phaA2phaB1-phaC1 and pETDuet1-phaA2phaB1-phaC1-vgb were identified as PHB by Fourier Transform Infrared and Proton nuclear magnetic resonance spectroscopy analysis. Furthermore, the decomposition temperature at 10% weight loss of PHB extracted from Massilia sp. UMI-21, recombinant E. coli DH5α-pETDuet1-phaA2phaB1-phaC1 and DH5α-pETDuet1-phaA2phaB1-phaC1-vgb was 276.5, 278.7 and 286.3 °C, respectively, showing good thermal stability. Herein, we presented the whole genome information of PHB-producing Massilia sp. UMI-21 and constructed novel recombinant strains using key genes in PHB synthesis of strain UMI-21 and the vgb gene. This genetically engineered E. coli strain can serve as an effective novel candidate in E. coli cell factory for PHB production by the rapid cell growth and high PHB production. The online version contains supplementary material available at 10.1186/s12934-023-02142-x.
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