水栖气单胞菌 4AK4 的代谢工程改造用于生产 3-羟基丁酸和中链长度 3-羟基烷酸的共聚物。
Metabolic engineering of Aeromonas hydrophila 4AK4 for production of copolymers of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoate.
机构信息
Protein Science Laboratory of the Ministry of Education, Department of Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
出版信息
Bioresour Technol. 2011 Sep;102(17):8123-9. doi: 10.1016/j.biortech.2011.05.074. Epub 2011 May 30.
A mutant termed Aeromonas hydrophila AKLF was constructed by deleting acetic acid pathway related genes pta and ackA in A. hydrophila 4AK4. Accumulation of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) in A. hydrophila AKLF was increased by 47% from 2.11 to 3.10g/L associated with a reduction on acetic acid formation compared with A. hydrophila 4AK4 when lauric acid was used as carbon resource. A. hydrophila AKLF harboring pVGAB encoding Vitreoscilla hemoglobin, β-ketothiolase and acetoacetyl-CoA reductase was found to produce 85% more PHBHHx compared to its wild type. Expression of plasmid pD(Ec)L(Pp) harboring genes related to fatty acid metabolism in A. hydrophila AKLF led to 63% more PHBHHx production than A. hydrophila 4AK4. Replacing phaC in A. hydrophila AKLF with a mutant phaC2 from Pseudomonas stutzeri 1317 resulted in enhanced production of copolymers of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates compared to A. hydrophila 4AK4 harboring the mutant phaC2 in the chromosome as control.
通过缺失嗜水气单胞菌 4AK4 中与乙酸途径相关的基因 pta 和 ackA，构建了一个突变体 Aeromonas hydrophila AKLF。与嗜水气单胞菌 4AK4 相比，当以月桂酸为碳源时，A. hydrophila AKLF 中聚（3-羟基丁酸-co-3-羟基己酸）（PHBHHx）的积累量增加了 47%，从 2.11g/L 增加到 3.10g/L，同时乙酸的形成减少。与野生型相比，携带编码威氏血红蛋白、β-酮硫解酶和乙酰乙酰辅酶 A 还原酶的 pVGAB 的 A. hydrophila AKLF 产生的 PHBHHx 增加了 85%。在 A. hydrophila AKLF 中表达与脂肪酸代谢相关的质粒 pD(Ec)L(Pp)，导致 PHBHHx 的产量比 A. hydrophila 4AK4 增加了 63%。用来自施氏假单胞菌 1317 的突变体 phaC2 替代 A. hydrophila AKLF 中的 phaC，与作为对照的染色体中携带突变体 phaC2 的 A. hydrophila 4AK4 相比，共聚物 3-羟基丁酸和中链长度 3-羟基烷酸的产量得到了增强。
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