Penicilliumsp.MYA5中microperfuranone聚酮合酶基因在构巢曲霉中的异源表达

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中图分类号：Q786 文献标志码：A

Heterologous expression of the microperfuranone PKS from Penicillium sp.MYA5 in Aspergillus nidulans

MengXianchao1，HuBo²，YuHaobing²，SunYuanyuan²，andLiuXiaoyu² （1 SchoolofOceanFoodandBilogicalEngieeingJangsuOceanUnversityLianungang2O5;2LANavalMedicalCeter Naval Medical University， Shanghai 200433）

Abstract Objective To enhance the production of the antimicrobial compound microperfuranone derived from the polar endophytic fungus Penicillium sp.MYA5 through gene mining and heterologous expression strategies， and to evaluate its bioactivity.MethodsGenome sequencing data for Penicillium sp.MYA5 were analyzed using antiSMASH to identify biosynthetic gene clusters.The polyketide synthase gene mfpksA wascloned and inserted into the recombinant plasmid pYTU-mfpksA.Screening positive clones through yeastand Escherichia coli transformation. The recombinant plasmid was introduced into Aspergillus nidulans LO8030，and screened on CHsorbitol plates to obtain therecombinant strainThe plasmid was transformed into Saccharomyces cerevisiaeBJ5464andEscherichia coli TOP10 forvalidation，heterologous expression was performed in Aspergilus nidulans LO8030，followed by fermentation， HPLC analysis，and structural elucidation via NMR and massspectrometry. Antimicrobial activity was assessed against pathogenic strains using the broth microdilution method. ResultsThe target compound microperfuranone was successfullyisolated from the heterologous expression strain，achieving anapproximately3.88- fold increase in yield compared to the MYA5 strain.Antimicrobial activity assays revealed that microperfuranone exhibited a minimum inhibitory concentration （MIC） of 16μg/mL against both Fusarium oxysporum and Vibrio harveyi. Notably，its inhibitory activity against F. oxysporum was significantly superior to chloramphenicol. Conclusion The heterologous expression strategy successfully enhanced the production of microperfuranone， confirming the effectiveness of biosynthetic gene cluster mining and heterologous expression in natural product discovery.The compound exhibited significant inhibitory activity against plant pathogens，providing a theoretical foundation for the antimicrobial application of polar microbial resources.

Key WordsMicroperfuranone; Penicillium sp.; PKS; Heterologous expression; Polyketide compounds

呋喃酮类化合物是一类含有氧原子的五元杂环芳香化合物，具有抗菌、抗氧化、抗炎和抗癌等多种生物活性[1-3]，已开发上市的药物有痢特灵、呋喃酮唑片等，真菌中已报道的呋喃酮类化合物多数都是通过聚酮合成酶（PKS）途径合成的，如asperfuranone，gregatinA等[4-5]，其不同的官能团具有不同的生物活性[]，microperfuranone是1个联苯呋喃酮化合物，其结构为5-羟基-4-苄基-3-苯基-2（5H）-呋喃酮，已从真菌Aspergilluscarneus，Aspergillussp.Y-12和AcremoniumpilosumF47等菌株中分离得到[7-10]，具有抗菌，抗病毒等生物活性[1]，但通过常规分离手段的产量相对较低，可以采取基因工程技术对其生物合成基因进行异源表达，不仅能避免原始宿主复杂代谢网络对产物合成的干扰，而且目标化合物的产量也会有所提高。（剩余12457字）