文献类型：期刊文献

英文题名：Raman spectroscopy provides a rapid, non-invasive method for quantitation of starch in live, unicellular microalgae

作者：Ji, Yuetong[1,2];He, Yuehui[1,2,3];Cui, Yanbin[4];Wang, Tingting[1,2];Wang, Yun[1,2];Li, Yuanguang[4];Huang, Wei E.[5];Xu, Jian[1,2]

机构：[1]Chinese Acad Sci, CAS Key Lab Biofuels, Single Cell Ctr, Qingdao, Shandong, Peoples R China;[2]Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, Shandong Key Lab Energy Genet, Qingdao, Shandong, Peoples R China;[3]Univ Chinese Acad Sci, Beijing, Peoples R China;[4]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[5]Univ Sheffield, Kroto Inst, Sheffield, S Yorkshire, England

年份：2014

卷号：9

期号：12

起止页码：1512

外文期刊名：BIOTECHNOLOGY JOURNAL

收录：;EI(收录号：201436028188);WOS:【SCI-EXPANDED(收录号:WOS:000346021200011)】；

基金：This work was supported by the National High-Tech Development Program (2012AA02A707) and National Basic Research Program of China (2012CB721101 and 2011CB200902) from Ministry of Science and Technology of China and by International Partnership Innovation Program from Chinese Academy of Sciences.

语种：英文

外文关键词：Microalgae; Non-invasive analysis; Raman spectroscopy; Single-cell analysis; Starch

摘要：Conventional methods for quantitation of starch content in cells generally involve starch extraction steps and are usually labor intensive, thus a rapid and non-invasive method will be valuable. Using the starch-producing unicellular microalga Chlamydomonas reinhardtii as a model, we employed a customized Raman spectrometer to capture the Raman spectra of individual single cells under distinct culture conditions and along various growth stages. The results revealed a nearly linear correlation (R-2 = 0.9893) between the signal intensity at 478 cm(-1) and the starch content of the cells. We validated the specific correlation by showing that the starch-associated Raman peaks were eliminated in a mutant strain where the AGPase (ADP-glucose pyrophosphorylase) gene was disrupted and consequentially the biosynthesis of starch blocked. Furthermore, the method was validated in an industrial algal strain of Chlorella pyrenoidosa. This is the first demonstration of starch quantitation in individual live cells. Compared to existing cellular starch quantitation methods, this single-cell Raman spectra-based approach is rapid, label-free, non-invasive, culture-independent, low-cost, and potentially able to simultaneously track multiple metabolites in individual live cells, therefore should enable many new applications.