Study on the Visualization of Pigment in Haematococcus pluvialis by Raman Spectroscopy Technique

As an ideal raw material for the production of astaxanthin, H. pluvialis was drawing attention during the last few years, there are some research topics initiated to find out the synthetic pathway of astaxanthin in H. pluvialis. In this study, confocal microscopic Raman technology was utilized to analyze the point-by-point mapping for H. pluvialis, and the visualization of pigment such as carotenoid and astaxanthin content were achieved. By comparing the Raman spectra of H. pluvialis and standard product of astaxanthin, and using the C = C stretching vibration of the Raman intensity as the main indicator for carotenoids, the visual spatial distribution for the carotenoids content was obtained. The MCR-ALS was applied to analyze the Raman data of H. pluvialis, and the information of astaxanthin was extracted to achieve real-time spatial distribution. The visualization of astaxanthin content shows that MCR-ALS is very effective for extracting the information of astaxanthin content from H. pluvialis. By exploring the spatial distribution of carotenoids and astaxanthin contents, analyzing the spatial distribution rules during its growth, Raman spectroscopy technology can be utilized to investigate the internal components of the pigment (ataxanthin, etc.) in H. pluvialis, which make it as an effective methodology to monitor the accumulation and changing mechanism of pigment content in microalgae.

[1]  T. Baldacchini,et al.  Coherent anti-Stokes Raman scattering and spontaneous Raman spectroscopy and microscopy of microalgae with nitrogen depletion , 2012, Biomedical optics express.

[2]  A. Hirata,et al.  Three-Dimensional Ultrastructural Study of Oil and Astaxanthin Accumulation during Encystment in the Green Alga Haematococcus pluvialis , 2013, PloS one.

[3]  A. Solovchenko Recent breakthroughs in the biology of astaxanthin accumulation by microalgal cell , 2015, Photosynthesis Research.

[4]  Howland D. T. Jones,et al.  Carotenoid Distribution in Living Cells of Haematococcus pluvialis (Chlorophyceae) , 2011, PloS one.

[5]  Venkatesh Balan,et al.  Designer synthetic media for studying microbial-catalyzed biofuel production , 2015, Biotechnology for Biofuels.

[6]  J. Mesquita,et al.  Ultrastructural study of Haematococcus lacustris (Girod.) Rostafinski (Volvocales). II: Mitosis and cytokinesis , 1984 .

[7]  Dor Ben-Amotz,et al.  Enhanced Chemical Classification of Raman Images in the Presence of Strong Fluorescence Interference , 2000 .

[8]  B. Kowalski,et al.  Multivariate curve resolution applied to spectral data from multiple runs of an industrial process , 1993 .

[9]  Z. Qiu,et al.  Microalgal detection by Raman microspectroscopy , 2014 .

[10]  D. Naumann,et al.  Identification of medically relevant microorganisms by vibrational spectroscopy. , 2002, Journal of microbiological methods.

[11]  B. Wood,et al.  A portable Raman acoustic levitation spectroscopic system for the identification and environmental monitoring of algal cells. , 2005, Analytical chemistry.

[12]  Jian Ling,et al.  Direct Raman imaging techniques for study of the subcellular distribution of a drug. , 2002, Applied optics.

[13]  K. Turnau,et al.  In situ Raman imaging of astaxanthin in a single microalgal cell. , 2011, The Analyst.

[14]  Romà Tauler,et al.  Application of multivariate self-modeling curve resolution to the quantitation of trace levels of organophosphorus pesticides in natural waters from interlaboratory studies , 1996 .

[15]  B. Wood,et al.  Effects of pre‐processing of Raman spectra on in vivo classification of nutrient status of microalgal cells , 2006 .

[16]  B. Lendl,et al.  Multidimensional information on the chemical composition of single bacterial cells by confocal Raman microspectroscopy. , 2000, Analytical chemistry.

[17]  Graciela M. Escandar,et al.  Multivariate Curve Resolution–Alternating Least-Squares , 2014 .

[18]  Jun Cheng,et al.  In vivo kinetics of lipids and astaxanthin evolution in Haematococcus pluvialis mutant under 15% CO2 using Raman microspectroscopy. , 2017, Bioresource technology.

[19]  Pavel Zemánek,et al.  Raman Microspectroscopy of Individual Algal Cells: Sensing Unsaturation of Storage Lipids in vivo , 2010, Sensors.

[20]  A. Kaczor,et al.  Structural changes of carotenoid astaxanthin in a single algal cell monitored in situ by Raman spectroscopy. , 2011, Analytical chemistry.

[21]  D. Nelson,et al.  An integrative Raman microscopy-based workflow for rapid in situ analysis of microalgal lipid bodies , 2015, Biotechnology for Biofuels.

[22]  K. Grünewald,et al.  Ketocarotenoid Biosynthesis Outside of Plastids in the Unicellular Green Alga Haematococcus pluvialis * , 2001, The Journal of Biological Chemistry.

[23]  Qing Huang,et al.  Screening of Astaxanthin-Hyperproducing Haematococcus pluvialis Using Fourier Transform Infrared (FT-IR) and Raman Microspectroscopy , 2016, Applied spectroscopy.

[24]  Mats Josefson,et al.  Characterization and mapping of carotenoids in the algae Dunaliella and Phaeodactylum using Raman and target orthogonal partial least squares , 2011 .

[25]  E. McHugh,et al.  Effect of various stress-regulatory factors on biomass and lipid production in microalga Haematococcus pluvialis. , 2013, Bioresource technology.

[26]  R. Goodacre,et al.  Provided for Non-commercial Research and Educational Use Only. Not for Reproduction, Distribution or Commercial Use. Shining Light on the Microbial World: the Application of Raman Microspectroscopy , 2022 .

[27]  Q. Hu,et al.  Consumption of oxygen by astaxanthin biosynthesis: a protective mechanism against oxidative stress in Haematococcus pluvialis (Chlorophyceae). , 2008, Journal of plant physiology.