Application of the cbh1 promoter in Mortierella alpina and optimisation of induction conditions,Letters in Applied Microbiology

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Application of the cbh1 promoter in Mortierella alpina and optimisation of induction conditions
Letters in Applied Microbiology ( IF 2.0 ) Pub Date : 2020-05-12 , DOI: 10.1111/lam.13300
Y Wang _{1,

2} , X Tang _{1,

2} , S Wang _{1,

2} , H Zhang _{1,

2,

3,

4} , Y Q Chen _{1,

2,

3,

4} , H Chen _{1,

2} , W Chen _{1,

2,

3,

5}

Affiliation

Mortierella alpina has gained remarkable interest due to its high capacity for arachidonic acid (AA) production and potential for eicosapentaenoic acid (EPA) production recently. However, the development of genetically modified strains is limited by lacking inducible promoters, which can express genes conditionally. Here the inducible promoter of cellobiohydrolase (Pcbh1) was utilized in M. alpina and the gene oPpFADS17 encoding ω‐3 fatty acid desaturase was selected as the reporter gene. Under conditions with inducer, expression of this gene enables M. alpina to produce EPA at room temperature, while no EPA was detected without inducer. We then optimized the induction conditions. The results demonstrated that the optimal induction condition was broth medium with 1% avicel as the inducer and 5% glucose as extra carbon source and the transcription level of the reporter gene was increasing with the extension of induction time. Successful application of Pcbh1 in M. alpina would significantly contribute to the steerable system to construct engineered strains for industrial production of microbial oils.

中文翻译：

cbh1启动子在高山被孢霉中的应用及诱导条件的优化

高山被孢霉最近因其花生四烯酸 (AA) 生产能力和二十碳五烯酸 (EPA) 生产潜力而引起了极大的兴趣。然而，转基因菌株的发展受到缺乏诱导型启动子的限制，这些启动子可以有条件地表达基因。在这里，纤维二糖水解酶（Pcbh1）的诱导型启动子被用于高山被孢霉，编码ω-3脂肪酸去饱和酶的基因oPpFADS17被选为报告基因。在有诱导剂的条件下，该基因的表达使高山被孢霉在室温下产生 EPA，而在没有诱导剂的情况下检测不到 EPA。然后我们优化了诱导条件。结果表明，最佳诱导条件为1%微晶纤维素为诱导剂，5%葡萄糖为额外碳源的肉汤培养基，报告基因的转录水平随着诱导时间的延长而增加。Pcbh1 在 M. alpina 中的成功应用将显着有助于构建用于微生物油工业生产的工程菌株的可控系统。

更新日期：2020-05-12

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