Renta Person Pesona, Huang Ta-Yu, Yu Ping-Hao, Jang Anna C-C, Chen Yi-Min
Department of Biotechnology and Bioindustry Sciences, National Cheng Kung University, Tainan, Taiwan.
AMB Express. 2025 Mar 20;15(1):52. doi: 10.1186/s13568-025-01859-9.
Aurantiochytrium are protists found in marine and estuarine environments, known for producing high quantities of omega-3 fatty acids, particularly docosahexaenoic acid (DHA). The commercial viability of this species is currently hindered by the lack of reliable screening methods for the rapid identification of strains with high DHA content. This study developed a high-throughput screening platform based on the sulfo-phospho-vanillin (SPV) reaction, which produces a pink chromophore upon reacting with C-C double bonds in lipids. Analysis of 200 strains derived through the UV mutagenesis of the Aurantiochytrium limacinum strain BL10 revealed 7 strains that exhibited significantly elevated SPV reactivity, compared to the naïve strain (P < 0.01). Gas chromatography-mass spectrometry analysis revealed that in 4 of the 7 strains, DHA levels were significantly higher than those of the naïve strain. Among those 4 strains, the SPV reactivity values of two strains were precisely correlated with their DHA/DPA content. The SPV reaction protocol proposed in this paper has considerable potential for the high-throughput screening of Aurantiochytrium strains particularly those derived through UV mutagenesis.
金藻是在海洋和河口环境中发现的原生生物，以产生大量的ω-3脂肪酸，特别是二十二碳六烯酸（DHA）而闻名。目前，该物种的商业可行性受到缺乏可靠筛选方法的阻碍，这些方法用于快速鉴定高DHA含量的菌株。本研究开发了一种基于磺基磷香草醛（SPV）反应的高通量筛选平台，该反应在与脂质中的碳-碳双键反应时会产生粉红色发色团。对通过对纤细金藻菌株BL10进行紫外线诱变获得的200个菌株进行分析，发现有7个菌株与原始菌株相比，SPV反应性显著提高（P < 0.01）。气相色谱-质谱分析表明，在这7个菌株中的4个菌株中，DHA水平显著高于原始菌株。在这4个菌株中，有两个菌株的SPV反应性值与它们的DHA/DPA含量精确相关。本文提出的SPV反应方案在高通量筛选金藻菌株，特别是通过紫外线诱变获得的菌株方面具有相当大的潜力。
