对具有较高异养细胞分裂率的血球藻(叶绿藻科)突变体进行蛋白质组学和磷酸蛋白质组学分析,揭示了细胞增殖和营养分配途径的改变。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
Kyarii Ramarui, Jun Zhong, Yantao Li
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引用次数: 0

摘要

血球藻已被用于生产酮类类胡萝卜素抗氧化剂虾青素。目前,H. pluvialis 的异养培养受到生长速度缓慢的限制。这项工作旨在通过探索血球菌的乙酸代谢机制来应对这一挑战。通过化学诱变和筛选发现,血球藻菌株 KREMS 23D-3 在醋酸异养生长时,细胞密度比野生型高出 34.9%。采用蛋白质组学和磷酸化蛋白质组学的综合方法,对野生型和突变株中的 4955 个蛋白质和 2505 个磷酸化蛋白质中的 5099 个磷酸化位点进行了量化。其中,突变体中有 12 个蛋白质明显上调,22 个明显下调,而磷酸化蛋白质组分析则在 106 个蛋白质上发现了 143 个明显上调的磷酸化位点,在 114 个蛋白质上发现了 130 个下调的磷酸化位点。突变体中无性繁殖促进复合磷蛋白的上调和一种推定的细胞周期分裂 20 磷蛋白的下调表明,有丝分裂进展迅速,同时细胞分裂率较高。突变体中上调的共卟啉原氧化酶和磷酸化镁螯合酶表明,叶绿素生物合成的氮分配发生了改变。大量不同表达的磷蛋白表明,磷酸化是血球菌蛋白质表达和活性的关键调节因子。总之,本研究揭示了血球柄菌中相互关联的醋酸代谢途径的调控,并提供了可指导未来菌株工程工作的蛋白质目标。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Proteomic and phosphoproteomic analysis of a Haematococcus pluvialis (Chlorophyceae) mutant with a higher heterotrophic cell division rate reveals altered pathways involved in cell proliferation and nutrient partitioning

Haematococcus pluvialis has been used to produce the ketocarotenoid antioxidant, astaxanthin. Currently, heterotrophic cultivation of H. pluvialis is limited by slow growth rates. This work aimed to address this challenge by exploring the mechanisms of acetate metabolism in Haematococcus. Chemical mutagenesis and screening identified H. pluvialis strain KREMS 23D-3 that achieved up to a 34.9% higher cell density than the wild type when grown heterotrophically on acetate. An integrative proteomics and phosphoproteomics approach was employed to quantify 4955 proteins and 5099 phosphorylation sites from 2505 phosphoproteins in the wild-type and mutant strains of H. pluvialis. Among them, 12 proteins were significantly upregulated and 22 significantly downregulated in the mutant while phosphoproteomic analysis identified 143 significantly upregulated phosphorylation sites on 106 proteins and 130 downregulated phosphorylation sites on 114 proteins. Upregulation of anaphase-promoting complex phosphoproteins and downregulation of a putative cell cycle division 20 phosphoprotein in the mutant suggests rapid mitotic progression, coinciding with higher cell division rates. Upregulated coproporphyrinogen oxidase and phosphorylated magnesium chelatase in the mutant demonstrated altered nitrogen partitioning toward chlorophyll biosynthesis. The large proportion of differentially expressed phosphoproteins suggests phosphorylation is a key regulator for protein expression and activity in Haematococcus. Taken together, this study reveals the regulation of interrelated acetate metabolic pathways in H. pluvialis and provides protein targets that may guide future strain engineering work.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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