Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering.

IF 4.3 2区 生物学 Q1 BIOLOGY Biological Research Pub Date : 2024-11-05 DOI:10.1186/s40659-024-00559-1
Alejandro Durán, Maximiliano Venegas, Salvador Barahona, Dionisia Sepúlveda, Marcelo Baeza, Víctor Cifuentes, Jennifer Alcaíno
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Abstract

The yeast Xanthophyllomyces dendrorhous synthesizes astaxanthin, a high-value carotenoid with biotechnological relevance in the nutraceutical and aquaculture industries. However, enhancing carotenoid production through strain engineering remains an ongoing challenge. Recent studies have demonstrated that carotenogenesis in X. dendrorhous is regulated by the SREBP pathway, which includes the transcription factor Sre1, particularly in the mevalonate pathway that also produces precursors used for ergosterol synthesis. In this study, we explored a novel approach to enhance carotenoid synthesis by replacing the native crtE promoter, which drives geranylgeranyl pyrophosphate synthesis (the step where carotenogenesis diverges from ergosterol biosynthesis), with the promoter of the HMGS gene, which encodes 3-hydroxy-3-methylglutaryl-CoA synthase from the mevalonate pathway. The impact of this substitution was evaluated in two mutant strains that already overproduce carotenoids due to the presence of an active Sre1 transcription factor: CBS.cyp61-, which does not produce ergosterol and strain CBS.SRE1N.FLAG, which constitutively expresses the active form of Sre1. Wild-type strain CBS6938 was used as a control. Our results showed that this modification increased the crtE transcript levels more than threefold and fourfold in CBS.cyp61-.pHMGS/crtE and CBS.SRE1N.FLAG.pHMGS/crtE, respectively, resulting in 1.43-fold and 1.22-fold increases in carotenoid production. In contrast, this modification did not produce significant changes in the wild-type strain, which lacks the active Sre1 transcription factor under the same culture conditions. This study highlights the potential of promoter substitution strategies involving genes regulated by Sre1 to enhance carotenoid production, specifically in strains where the SREBP pathway is activated, offering a promising avenue for strain improvement in industrial applications.

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提高黄ophyllomyces dendrorhous/Phaffia rhodozyma 的类胡萝卜素产量:SREBP 通路激活和启动子工程。
黄ophyllomyces dendrorhous酵母能合成虾青素,这是一种高价值的类胡萝卜素,在营养保健品和水产养殖业中具有重要的生物技术意义。然而,通过菌种工程提高类胡萝卜素的产量仍然是一个持续的挑战。最近的研究表明,X. dendrorhous 的类胡萝卜素生成受 SREBP 通路调控,其中包括转录因子 Sre1,尤其是在甲羟戊酸通路中,该通路还产生用于麦角甾醇合成的前体。在本研究中,我们探索了一种增强类胡萝卜素合成的新方法,即用甲羟戊酸途径中编码 3-羟基-3-甲基戊二酰-CoA 合成酶的 HMGS 基因的启动子取代原生的 crtE 启动子,后者驱动香叶基焦磷酸酯的合成(胡萝卜素生成与麦角甾醇生物合成分化的步骤)。我们在两个突变菌株中评估了这一替换的影响,这两个突变菌株由于存在活性 Sre1 转录因子,已经过量产生类胡萝卜素:这两个菌株分别是不产生麦角甾醇的 CBS.cyp61-和组成型表达 Sre1 活性形式的菌株 CBS.SRE1N.FLAG。野生型菌株 CBS6938 用作对照。我们的结果表明,这种修饰使 CBS.cyp61-.pHMGS/crtE 和 CBS.SRE1N.FLAG.pHMGS/crtE 中的 crtE 转录水平分别提高了三倍和四倍多,导致类胡萝卜素产量分别增加了 1.43 倍和 1.22 倍。相比之下,在相同的培养条件下,缺乏活性 Sre1 转录因子的野生型菌株的这种修饰并没有产生显著变化。这项研究强调了涉及受 Sre1 调控基因的启动子替换策略在提高类胡萝卜素产量方面的潜力,特别是在 SREBP 途径被激活的菌株中,这为工业应用中的菌株改良提供了一条很有前景的途径。
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来源期刊
Biological Research
Biological Research 生物-生物学
CiteScore
10.10
自引率
0.00%
发文量
33
审稿时长
>12 weeks
期刊介绍: Biological Research is an open access, peer-reviewed journal that encompasses diverse fields of experimental biology, such as biochemistry, bioinformatics, biotechnology, cell biology, cancer, chemical biology, developmental biology, evolutionary biology, genetics, genomics, immunology, marine biology, microbiology, molecular biology, neuroscience, plant biology, physiology, stem cell research, structural biology and systems biology.
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