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AplaeA 在出芽短梗霉中孢子生成和聚苹果酸合成的调控作用。

Role of AplaeA in the regulation of spore production and poly(malic acid) synthesis in Aureobasidium pullulans.

机构信息

School of Life Science and Technology, Harbin Institute of Technology, Harbin 150006, China.

School of Life Science and Technology, Harbin Institute of Technology, Harbin 150006, China; State Key Laboratory of Urban Water Resources and Environment, Harbin Institute of Technology, Harbin 150090, China.

出版信息

Int J Biol Macromol. 2024 Nov;279(Pt 1):135153. doi: 10.1016/j.ijbiomac.2024.135153. Epub 2024 Aug 28.

Abstract

Poly(malic acid) (PMLA) as one of the important metabolites in Aureobasidium pullulans has great application in a variety of fields. LaeA, a global regulator capable of controlling fungal secondary metabolites, has been identified in A. pullulans. The involvement of LaeA in the biosynthesis of PMLA through direct or indirect means and its role in A. pullulans is currently unknown. Here we extracted and characterized the AplaeA gene from A. pullulans HIT-LCY3 and resolved it by bioinformatics. The function of AplaeA was subsequently characterized by Rnai-LaeA and OEX-LaeA mutants. Based on phenotypic observations of the mutant strains, it was found that the gene had a large effect on the morphology and melanin production aspects of the strain, and that overexpression of the AplaeA gene resulted in a substantial increase in spore numbers. RT-qPCR combined with protein interactions results analyzed that ApLaeA positively regulates PMLA biosynthesis by controlling the expression of core genes of PMLA biosynthesis gene cluster and other related regulatory factors. Finally, OEX-LaeA was used as the starting strain to obtain the optimal fermentation conditions. These findings illustrate the regulatory mechanism of a hypothesized global regulator, LaeA, in A. pullulans HIT-LCY3, suggesting its potential application in industrial-scale PMLA biosynthesis.

摘要

聚苹果酸(PMLA)是出芽短梗霉中重要的代谢产物之一,在许多领域具有广泛的应用。LaeA 是一种能够控制真菌次生代谢物的全局调控因子,在出芽短梗霉中已经被鉴定出来。LaeA 通过直接或间接的方式参与 PMLA 的生物合成,以及它在出芽短梗霉中的作用目前尚不清楚。本研究从出芽短梗霉 HIT-LCY3 中提取并鉴定了 AplaeA 基因,并通过生物信息学进行了解析。随后通过 Rnai-LaeA 和 OEX-LaeA 突变体对 AplaeA 的功能进行了表征。基于突变菌株的表型观察,发现该基因对菌株的形态和黑色素产生方面有很大的影响,并且 AplaeA 基因的过表达导致孢子数量显著增加。RT-qPCR 结合蛋白相互作用结果分析表明,ApLaeA 通过控制 PMLA 生物合成基因簇的核心基因和其他相关调控因子的表达来正向调控 PMLA 生物合成。最后,以 OEX-LaeA 为出发菌株,获得了最佳发酵条件。这些发现说明了假设的全局调控因子 LaeA 在出芽短梗霉 HIT-LCY3 中的调控机制,表明其在工业规模的 PMLA 生物合成中具有潜在的应用价值。

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