广藿香醇合成酶分子改造及重组大肠杆菌发酵合成广藿香醇
Molecular modification of patchoulol synthase and patchoulol production in recombinant Escherichia coli
广藿香醇是一种天然的三环倍半萜化合物，在日化品领域应用广泛。本研究对生物合成广藿香醇的关键酶--广藿香醇合成酶（PTS）的野生变种进行筛选，改造PTS3底物结合位点附近氨基酸残基，并通过同源序列比对改造差异位点，构建了组合突变体PTS3mut4。含有该突变体的重组大肠杆菌菌株，其广藿香醇摇瓶发酵产量比PTS3提高了1.01倍，达到448 mg/L，得率达到39.1 mg/g DCW，体积生产强度达到112 mg/L/d。在5 L发酵罐中，发酵168 h，广藿香醇产量可达2128 mg/L，体积生产强度达到304 mg/L/d。该结果对深入揭示PTS催化机制以及工业化发酵合成广藿香醇具有促进作用。
生物工程学分子生物学生物化学
发酵工程广藿香醇广藿香醇合成酶定点突变大肠杆菌
张新蕊,沈佳雯,杨诗怡,李云燕,刘海丽,王沁,蒋紫怡,周哲敏,王梦轩,周丽,黎君,张慧.广藿香醇合成酶分子改造及重组大肠杆菌发酵合成广藿香醇[EB/OL].(2023-02-03)[2025-10-10].http://www.paper.edu.cn/releasepaper/content/202302-19.点此复制
Patchoulol is a natural tricyclic sesquiterpene compound, which is widely used in the field of daily chemical products. In this study, the natural variant of patchoulol synthase (PTS), a key enzyme for biosynthesis of patchoulol, was screened. A combined mutant of PTS3mut4 was obtained by modifying the amino acid residues near the binding site of the substrate in PTS3 and the differential sites selected by homologous sequence alignment. The recombinant Escherichia coli strain containing PTS3mut4 produced 2.01 times higher titer (448 mg/L) than that of PTS3, with yield of 39.1 mg/g DCW and volume productivity of 112 mg/L/d shake-flask experiment. In a 5-L bioreactor, the yield of patchoulol reached 2128 mg/L, and the volume productivity reached 304 mg/L/d in 168 h. These results are helpful for understanding the catalytic mechanism of PTS and the industrial production of patchoulol.
Fermentation engineeringPatchoulolPatchoulol synthaseSite-directed mutagenesisEscherichia coli
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