An attempt to simultaneously quantify the polysaccharide, total lipid, protein and pigment in single Cyclotella cryptica cell by Raman spectroscopy.
An attempt to simultaneously quantify the polysaccharide, total lipid, protein and pigment in single Cyclotella cryptica cell by Raman spectroscopy.
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DOI：
10.1186/s13068-023-02314-2
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发表时间：
2023-04-08
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--
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目前，传统的测定微藻光合作用产物的方法通常是基于大量的细胞质量达到测量基线，结果只能揭示种群水平上的平均状态，这对于从大量潜在的微藻突变体中大规模、快速地筛选特定表型是不可行的。近年来，单细胞拉曼光谱(SCRS)被证明能够快速同时定量微藻的生化成分。然而，该方法用于分析隐环藻(C.cryptica)的生化成分尚未见报道。因此，首次尝试用SCRS对该硅藻中的这四种生化成分进行测定。建立了同时测定隐藻中多糖、总脂、蛋白质和Chl-a含量的SCRS方法，其中13条谱带是硅藻成分的主要标记带。此外，基于全谱的偏最小二乘回归模型能够可靠地预测这四种细胞成分，其皮尔逊相关系数分别达到0.949、0.904、0.801和0.917。最后，基于一个同基因样本的SCRS数据，通过拉曼体内相关分析(IRCA)分析了这些成分之间的成对相关性和动态转化过程，结果表明，硅饥饿可以促进隐藻细胞中的碳从蛋白质和色素代谢流向多糖和脂肪代谢。首先，建立了隐藻单个细胞中多糖、总脂、蛋白质和色素的同时含量测定方法。其次，通过IRCA构建了细胞内组分的即时相互转换，该方法基于一个等基因群体的数据集，具有更高的准确性和时效性。结果表明，缺硅能促进隐藻细胞中的碳从蛋白质和色素代谢流向多糖和脂肪代谢。
At present, the conventional methods for determining photosynthetic products of microalgae are usually based on a large number of cell mass to reach the measurement baseline, and the result can only reveal the average state at the population level, which is not feasible for large-scale and rapid screening of specific phenotypes from a large number of potential microalgae mutants. In recent years, single-cell Raman spectra (SCRS) has been proved to be able to rapidly and simultaneously quantify the biochemical components of microalgae. However, this method has not been reported to analyze the biochemical components of Cyclotella cryptica (C. cryptica). Thus, SCRS was first attempt to determine these four biochemical components in this diatom. The method based on SCRS was established to simultaneously quantify the contents of polysaccharide, total lipids, protein and Chl-a in C. cryptica, with thirteen Raman bands were found to be the main marker bands for the diatom components. Moreover, Partial Least Square Regression (PLSR) models based on full spectrum can reliably predict these four cellular components, with Pearson correlation coefficient for these components reached 0.949, 0.904, 0.801 and 0.917, respectively. Finally, based on SCRS data of one isogenic sample, the pairwise correlation and dynamic transformation process of these components can be analyzed by Intra-ramanome Correlation Analysis (IRCA), and the results showed silicon starvation could promote the carbon in C. cryptica cells to flow from protein and pigment metabolism to polysaccharide and lipid metabolism. First, method for the simultaneous quantification of the polysaccharide, total lipid, protein and pigment in single C. cryptica cell are established. Second, the instant interconversion of intracellular components was constructed through IRCA, which is based on data set of one isogenic population and more precision and timeliness. Finally, total results indicated that silicon deficiency could promote the carbon in C. cryptica cells to flow from protein and pigment metabolism to polysaccharide and lipid metabolism.
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影响因子：
9
作者：
Maity, Jyoti Prakash;Bundschuh, Jochen;Bhattacharya, Prosun
通讯作者：
Bhattacharya, Prosun
影响因子：
2.9
作者：
HARRISON, PJ;WATERS, RE;TAYLOR, FJR
通讯作者：
TAYLOR, FJR
影响因子：
5.4
作者：
Guo B;Liu B;Yang B;Sun P;Lu X;Liu J;Chen F
通讯作者：
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影响因子：
12.3
作者：
Budnik B;Levy E;Harmange G;Slavov N
通讯作者：
Slavov N
影响因子：
5
作者：
Nemes, Peter;Knolhoff, Ann M.;Rubakhin, Stanislav S.;Sweedler, Jonathan V.
通讯作者：
Sweedler, Jonathan V.