Name: spectrophotometric determination of phycobiliprotein content in cyanobacterium synechocystis
Uploaded: 2018-09-11T13:30:00.000+00:00
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该议定书是从上一个出版物11通过的。t.z.、d. Č和 J. 由捷克共和国教育、青年和体育部在国家可持续发展方案 (NPU 一) 的支持下, 授予编号 LO1415。J. Č也得到了 GA CR 的支持, 18-24397S 号。捷克的系统生物学 C4SYS 研究基础设施 (项目没有 LM2015055) 支持获得仪器和其他设施的机会。m.a.s. 得到了俄罗斯科学基金会的资助 [14-14-00904]。

1. 蓝藻栽培

<ol><li>培养Synechocystis细胞在锥形烧瓶或光生物反应器10,19在缓冲 BG11 中等20保持 pH < 10 (例如, 使用17毫米 HEPES10)。 注: 标准栽培条件要求受控温度 (通常为30摄氏度, 最佳温度为35°c)21, 光照 (通常是强度高达800µmol [光子]/[m2·s])21的白色光, 和 CO2供应 (在400毫升平板光生物反应器, 增长饱和 CO2集中是 1700 ppm)21。</li>
	<li>要确定培养密度, 测量的文化光学密度 spectrophotometrically 在730毫微米 (OD730) 使用一个试管以 1 cm 的光路径. 或者, 使用 hemocytometer 或自动单元格在显微镜下计数细胞计数器。</li>
</ol>2. 样品准备
<ol><li>低辐照度下工作, 防止蛋白降解。</li>
	<li>收获 3 x 1 毫升的文化悬浮到安全锁管。使用 triplicates 来估计测量中的技术误差。在无菌条件下进行培养取样, 在层流罩中收获细胞, 并遵循适当的工作和安全做法。</li>
	<li>在实验室温度下将细胞在 1.5万 x g处离心成5分钟. 注意适当平衡的离心机转子。离心后, 丢弃上清液。一定不要打扰球团。</li>
	<li>把样品放在冰箱里。要长期贮存, 请将样品保持在摄氏-80 摄氏度。这是防止蛋白退化的必要措施。对于短期储存,-20 °c 是足够的。</li>
	<li>冷冻干燥样品过夜。在适当的冷冻干燥过程中, 将冷冻干燥冷凝器的温度保持在-60 摄氏度以下, 冷冻干燥室的压力约为1帕。</li>
	<li>完成冷冻干燥循环后, 应尽快关闭管, 以防止吸收的空气中的水。</li>
</ol>3. 细胞均匀化和色素提取
<ol><li>将四片玻璃珠 (直径2毫米) 添加到每个取样管上, 然后关闭管。 注: 当用于均匀化的安全锁管的盖子太薄时, 可以在匀质过程中断裂;因此, 本协议只推荐有强盖的安全锁管。</li>
	<li>融汇样品与玻璃珠十五年代在一个均匀机在实验室温度。 注: 在安全锁管的整个内表面均有适当的匀质试样。</li>
	<li>添加1毫升的 PBS 缓冲器 (pH 7.4), 预换热器到4摄氏度, 以提取藻胆蛋白的样品。 注意: 从这个步骤上, 保持样品在冰上, 以防止降解的提取蛋白。这是至关重要的。</li>
	<li>在实验室温度下, 将样品与 PBS 混合5秒。 注: 混合后, 样品呈绿色。</li>
	<li>混合后, 将样品放在冰上60分钟, 用盖子盖住冰浴, 以防颜料降解。</li>
	<li>经过60分钟的蛋白萃取, 离心机的样品在 1.5万 x克4 摄氏度为5分钟. 注意适当平衡离心机转子。 注: 离心后, 上清有青色蓝色。</li>
</ol>4. 蛋白量化
<ol><li>在分光光度测量之前, 用 PBS 缓冲器将光谱仪校准为空白。</li>
	<li>校准分光光度计后, 将 PBS 从一个分光光度计试管, 用提取的藻胆蛋白代替所丢弃的缓冲器, 将上清液吸管移除。</li>
	<li>用 0.5 nm 的狭缝宽度量化蛋白浓度 spectrophotometrically。<ol>
<li>分别测量藻和复方阿司匹林中 phycocyanobilins 在 615 nm (615) 和 652 nm (652) 的 PBS 缓冲器上的吸光度, 并测量在 720 nm (720) 的细胞碎片的吸收量。</li>
		<li>根据班尼特和 Bogorad12的方程式 (1) 和 (2) 计算藻和复方阿司匹林的浓度: <img alt="Equation 1" src="/files/ftp_upload/58076/58076eq1.jpg"> <img alt="Equation 2" src="/files/ftp_upload/58076/58076eq2.jpg"> 注:615、652和720应符合分光光度计的线性吸收范围。如有必要, 用 PBS 缓冲器稀释样品。</li>
	</ol>
</li>
	<li>重新计算原始样品中的颜料浓度。样品中的颜料浓度与方程 (1) 和 (2) 的结果直接对应, 当1毫升的培养基和1毫升的萃取缓冲器用于分析时。如果使用不同数量的蓝藻样品和/或 PBS 缓冲器, 最后的颜料浓度应根据等式 (3) 计算: <img alt="Equation 3" src="/files/ftp_upload/58076/58076eq3.jpg">(3) 注: 在每细胞干重的正则蛋白含量的情况下, 应根据方程式 (4)<img alt="Equation 4" src="/files/ftp_upload/58076/58076eq4.jpg"> </img> (4) 计算最终颜料浓度。</li>
</ol>5. 测定细胞干重 (可选)
<ol><li>在分析天平上重三个空的安全锁管。 注意: 安全锁管干燥是至关重要的。如果将管子贮存在潮湿的环境中, 将管子在冷冻干燥机中干燥几个小时, 然后再称量。轻轻地操纵管子, 只用无粉手套的手, 以避免材料和研究员的手指之间的任何接触。保持所有的持有者和离心机的清洁, 以避免任何材料转移到管。</li>
	<li>样品 1 x 15 毫升的文化悬浮到15毫升圆锥管。</li>
	<li>离心机的培养悬浮在15毫升圆锥管在 4000 x g在实验室温度为10分钟. 平衡离心机转子与三额外15毫升圆锥管, 每个包含15毫升的水。离心后, 丢弃12毫升的上清。</li>
	<li>并用重悬颗粒在剩余的上清和转移 3 x 1 毫升混合物到三1.5 毫升安全锁管子与吸管。如果剩余的颗粒留在15毫升圆锥管, 添加1.5 毫升的去离子水到锥形管, 涡流或摇动管并用重悬余下的颗粒, 并转移 3 x 0.5 毫升的混合物到三1.5 毫升安全锁管 (已经 containing 3 x 1 毫升的颗粒) 与吸管。 注: 将球团除以三1.5 毫升的安全锁管, 可以估计干重测量的任何技术误差。</li>
	<li>在实验室温度为5分钟, 将1.5 毫升安全锁管中的细胞离心成 1.5万 x克. 平衡离心机转子与另外1.5 毫升安全锁管, 包含1毫升水。离心后, 丢弃上清液。</li>
	<li>把样品放在冰箱里。长期贮存, 样品保持在摄氏-80 摄氏度;为短期存贮,-20 °c 是充足的。</li>
	<li>冷冻干燥样品过夜。为适当的冷冻干燥过程, 保持冷冻干燥冷凝器的温度低于-90 °c 和在冷冻干燥室的压力约 1 hPa。</li>
	<li>冷冻干燥后, 关闭管, 并权衡样品的分析平衡。 注: 干重值可用于每细胞干重蛋白含量的规范化。</li>
</ol>

<ol>
	<li>Mimuro, M., Kikuchi, H., Green, B. R., Parson, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antenna+Systems+and+Energy+Transfer+in+Cyanophyta+and+Rhodophyta.">Antenna Systems and Energy Transfer in Cyanophyta and Rhodophyta.</a> Light-Harvesting Antennas in Photosynthesis. , 281-306 (2003).</li><li>Spear-bernstein, L., Miller, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unique+location+of+the+phycobiliprotein+light-harvesting+pigment+in+the+Cryptophyceae.">Unique location of the phycobiliprotein light-harvesting pigment in the Cryptophyceae.</a> Journal of Phycology. 25 (3), 412-419 (1989).</li><li>Kirst, H., Formighieri, C., Melis, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maximizing+photosynthetic+efficiency+and+culture+productivity+in+cyanobacteria+upon+minimizing+the+phycobilisome+light-harvesting+antenna+size.">Maximizing photosynthetic efficiency and culture productivity in cyanobacteria upon minimizing the phycobilisome light-harvesting antenna size.</a> Biochimica et Biophysica Acta - Bioenergetics. 1837 (10), 1653-1664 (2014).</li><li>Page, L. E., Liberton, M., Pakrasi, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+photoautotrophic+productivity+in+the+cyanobacterium+Synechocystis+sp.+strain+PCC+6803+by+phycobilisome+antenna+truncation.">Reduction of photoautotrophic productivity in the cyanobacterium Synechocystis sp. strain PCC 6803 by phycobilisome antenna truncation.</a> Applied and Environmental Microbiology. 78 (17), 6349-6351 (2012).</li><li>Sonani, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+production,+purification+and+applications+of+phycobiliproteins.">Recent advances in production, purification and applications of phycobiliproteins.</a> World Journal of Biological Chemistry. 7 (1), 100 (2016).</li><li>Bryant, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Molecular Biology of Cyanobacteria. , (1994).</li><li>Kaneko, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequence+analysis+of+the+genome+of+the+unicellular+cyanobacterium+Synechocystis+sp.+strain+PCC6803.+I.+Sequence+features+in+the+1+Mb+region+from+map+positions+64%+to+92%+of+the+genome.">Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome.</a> DNA Research. 2, 191-198 (1995).</li><li>Kaneko, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequence+analysis+of+the+genome+of+the+unicellular+cyanobacterium+Synechocystis+sp.+strain+PCC6803.+II.+Sequence+determination+of+the+entire+genome+and+assignment+of+potential+protein-coding+regions.">Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions.</a> DNA Research. 3, 109-136 (1996).</li><li>Grigorieva, G., Shestakov, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+in+the+cyanobacterium+Synechocystis+sp+6803.">Transformation in the cyanobacterium Synechocystis sp 6803.</a> FEMS Microbiology Letters. 13 (4), 367-370 (1982).</li><li>Zav&#345;el, T., Sinetova, M. A., B&#250;zov&#225;, D., Liter&#225;kov&#225;, P., &#268;erven&#253;, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+model+cyanobacterium+Synechocystis+sp:+PCC+6803+autotrophic+growth+in+a+flat-panel+photobioreactor.">Characterization of a model cyanobacterium Synechocystis sp: PCC 6803 autotrophic growth in a flat-panel photobioreactor.</a> Engineering in Life Sciences. 15 (1), (2015).</li><li>Zav&#345;el, T., O&#269;en&#225;&#353;ov&#225;, P., &#268;erven&#253;, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+characterization+of+Synechocystis+sp.+PCC+6803+substrains+reveals+differences+in+sensitivity+to+abiotic+stress.">Phenotypic characterization of Synechocystis sp. PCC 6803 substrains reveals differences in sensitivity to abiotic stress.</a> PLoS One. 12 (12), e0189130 (2017).</li><li>Bennett, A., Bogorad, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complementary+chromatic+adaption+in+a+filamentous+blue-green+alga.">Complementary chromatic adaption in a filamentous blue-green alga.</a> The Journal of Cell Biology. 58, 419-435 (1973).</li><li>L&#252;der, U. H., Knoetzel, J., Wiencke, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acclimation+of+photosynthesis+and+pigments+to+seasonally+changing+light+conditions+in+the+endemic+antarctic+red+macroalga+Palmaria+decipiens.">Acclimation of photosynthesis and pigments to seasonally changing light conditions in the endemic antarctic red macroalga Palmaria decipiens.</a> Polar Biology. 24 (8), 598-603 (2001).</li><li>Evans, L. V., Lobban, C. S., Chapman, D. J., Kremer, B. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+spectral+composition+and+irradiance+level+on+pigment+levels+in+seaweeds.">The effects of spectral composition and irradiance level on pigment levels in seaweeds.</a> Experimental Phycology: A Laboratory Manual. , 123-133 (1988).</li><li>Sampath-Wiley, P., Neefus, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+method+for+estimating+R-phycoerythrin+and+R-phycocyanin+contents+from+crude+aqueous+extracts+of+Porphyra+(Bangiales,+Rhodophyta).">An improved method for estimating R-phycoerythrin and R-phycocyanin contents from crude aqueous extracts of Porphyra (Bangiales, Rhodophyta).</a> Journal of Applied Phycology. 19 (2), 123-129 (2007).</li><li>Chung, Y. H., Park, Y. M., Moon, Y. J., Lee, E. M., Choi, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photokinesis+of+Cyanobacterium+Synechocystis+sp.+PCC+6803.">Photokinesis of Cyanobacterium Synechocystis sp. PCC 6803.</a> Journal of Photoscience. 11 (3), 89-94 (2004).</li><li>Sun, L., Gault, P. M., Marler, H. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phycobilisomes+from+Cyanobacteria.">Phycobilisomes from Cyanobacteria.</a> Handbook on Cyanobacteria: Biochemistry, Biotechnology and Applications. , 105-160 (2009).</li><li>Six, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diversity+and+evolution+of+phycobilisomes+in+marine+Synechococcus+spp.:+A+comparative+genomics+study.">Diversity and evolution of phycobilisomes in marine Synechococcus spp.: A comparative genomics study.</a> Genome Biology. 8 (12), (2007).</li><li>Sinetova, M. A., &#268;erven&#253;, J., Zav&#345;el, T., Nedbal, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+dynamics+and+constraints+of+batch+culture+growth+of+the+cyanobacterium+Cyanothece+sp.+ATCC+51142.">On the dynamics and constraints of batch culture growth of the cyanobacterium Cyanothece sp. ATCC 51142.</a> Journal of Biotechnology. 162 (1), (2012).</li><li>Stanier, R. Y., Kunisawa, R., Mandel, M., Cohen-Bazire, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purification+and+properties+of+unicellular+blue-green+algae+(order+Chroococcales).">Purification and properties of unicellular blue-green algae (order Chroococcales).</a> Bacteriological Reviews. 35 (2), 171-205 (1971).</li><li>Zav&#345;el, T., Sinetova, M. A., B&#250;zov&#225;, D., Liter&#225;kov&#225;, P., &#268;erven&#253;, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+model+cyanobacterium+Synechocystis+sp.+PCC+6803+autotrophic+growth+in+a+flat-panel+photobioreactor.">Characterization of a model cyanobacterium Synechocystis sp. PCC 6803 autotrophic growth in a flat-panel photobioreactor.</a> Engineering in Life Sciences. 15 (1), 122-132 (2015).</li><li>Hemlata, G., Fareha, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+Anabaena+sp.+nccu-9+with+special+reference+to+phycocyanin.">Studies on Anabaena sp. nccu-9 with special reference to phycocyanin.</a> Journal of Algal Biomass Utilization. 2 (1), 30-51 (2011).</li><li>Rito-Palomares, M., Nuez, L., Amador, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Practical+application+of+aqueous+two-phase+systems+for+the+development+of+a+prototype+process+for+c-phycocyanin+recovery+from+Spirulina+maxima.">Practical application of aqueous two-phase systems for the development of a prototype process for c-phycocyanin recovery from Spirulina maxima.</a> Journal of Chemical Technology &amp; Biotechnology. 76 (12), 1273-1280 (2001).</li><li>Zhang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selenium-Containing+Allophycocyanin+Purified+from+Selenium-Enriched+Spirulina+platensis+Attenuates+AAPH-Induced+Oxidative+Stress+in+Human+Erythrocytes+through+Inhibition+of+ROS+Generation.">Selenium-Containing Allophycocyanin Purified from Selenium-Enriched Spirulina platensis Attenuates AAPH-Induced Oxidative Stress in Human Erythrocytes through Inhibition of ROS Generation.</a> Journal of Agricultural and Food Chemistry. 59 (16), 8683-8690 (2011).</li><li>Nedbal, L., Trt&#237;lek, M., Cerven&#253;, J., Kom&#225;rek, O., Pakrasi, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+photobioreactor+system+for+precision+cultivation+of+photoautotrophic+microorganisms+and+for+high-content+analysis+of+suspension+dynamics.">A photobioreactor system for precision cultivation of photoautotrophic microorganisms and for high-content analysis of suspension dynamics.</a> Biotechnology and Bioengineering. 100 (5), 902-910 (2008).</li><li>Zav&#345;el, T., Knoop, H., Steuer, R., Jones, P. R., &#268;erven&#253;, J., Trt&#237;lek, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+evaluation+of+ethylene+production+in+the+recombinant+cyanobacterium+Synechocystis+sp.+PCC+6803+harboring+the+ethylene-forming+enzyme+by+membrane+inlet+mass+spectrometry.">A quantitative evaluation of ethylene production in the recombinant cyanobacterium Synechocystis sp. PCC 6803 harboring the ethylene-forming enzyme by membrane inlet mass spectrometry.</a> Bioresource Technology. 202, 142-151 (2016).</li><li>Smith, P. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+protein+using+bicinchoninic+acid.">Measurement of protein using bicinchoninic acid.</a> Analytical Biochemistry. 150 (1), 76-85 (1985).</li><li>Lawrenz, E., Fedewa, E. J., Richardson, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extraction+protocols+for+the+quantification+of+phycobilins+in+aqueous+phytoplankton+extracts.">Extraction protocols for the quantification of phycobilins in aqueous phytoplankton extracts.</a> Journal of Applied Phycology. 23 (5), 865-871 (2011).</li><li>Lea-Smith, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phycobilisome-Deficient+Strains+of+Synechocystis+sp.+PCC+6803+Have+Reduced+Size+and+Require+Carbon-Limiting+Conditions+to+Exhibit+Enhanced+Productivity.">Phycobilisome-Deficient Strains of Synechocystis sp. PCC 6803 Have Reduced Size and Require Carbon-Limiting Conditions to Exhibit Enhanced Productivity.</a> Plant Physiology. 165 (2), 705-714 (2014).</li><li>Seo, Y. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+isolation+of+phycocyanin+from+Spirulina+platensis+associated+with+high-pressure+extraction+process.">Stable isolation of phycocyanin from Spirulina platensis associated with high-pressure extraction process.</a> International Journal of Molecular Sciences. 14 (1), 1778-1787 (2013).</li><li>Touloupakis, E., Cicchi, B., Torzillo, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+bioenergetic+assessment+of+photosynthetic+growth+of+Synechocystis+sp.+PCC+6803+in+continuous+cultures.">A bioenergetic assessment of photosynthetic growth of Synechocystis sp. PCC 6803 in continuous cultures.</a> Biotechnology for Biofuels. 8 (1), 133 (2015).</li><li>Touloupakis, E., Cicchi, B., Benavides, A. M. S., Torzillo, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+high+pH+on+growth+of+Synechocystis+sp.+PCC+6803+cultures+and+their+contamination+by+golden+algae+(Poterioochromonas+sp.).">Effect of high pH on growth of Synechocystis sp. PCC 6803 cultures and their contamination by golden algae (Poterioochromonas sp.).</a> Applied Microbiology and Biotechnology. 100 (3), 1333-1341 (2016).</li><li>Ishii, A., Hihara, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+AbrB-Like+Transcriptional+Regulator,+Sll0822,+Is+Essential+for+the+Activation+of+Nitrogen-Regulated+Genes+in+Synechocystis+sp.+PCC+6803.">An AbrB-Like Transcriptional Regulator, Sll0822, Is Essential for the Activation of Nitrogen-Regulated Genes in Synechocystis sp. PCC 6803.</a> Plant Physiology. 148 (1), 660-670 (2008).</li></ol>

作者没有什么可透露的。

该协议描述了一种简单、快速、可重现的方法, 用于量化蓝藻Synechocystis中的蛋白内容。比较了细胞均匀化、蛋白质提取、藻和复方阿司匹林量化的几种方法, 最后的协议代表了每一个过程的最优步骤的组合。作为代表性数据, 藻胆蛋白在Synechocystis细胞中的含量随着光照强度的增加而定量化。尽管分析需要类似的时间和实验室设备作为一些以前发布的方法12,<su...

fPhycobiliproteins 是水溶性色素蛋白复合物, 代表原核蓝藻 (蓝藻) 和几种真核生物群中的光收获触角的主要成分 (Glaucophyta、红藻门和Cryptophyta)1。它们主要发生在称为 phycobilisomes 的超分子复合体上, 通常附着在基质侧的光合膜表面, 除Cryptophyta外, 藻胆蛋白在囊体腔2。迄今已确定了四种类型的藻胆蛋白: 核心复方阿司匹林和外围藻、藻红蛋白和 phycoerythrocyanin1。phycobilisomes 作为主要的光收获配合物, 是藻类和蓝藻群体培养生产力的重要因素之一。结果表明, phycobilisomes 截断可提高强光下3的生物量积累。另一方面, 在适度或低辐照度下, 天线截断导致生长速率和生物量积累减少3,4。藻胆蛋白在化妆品行业中作为食品着色剂、药品和食品添加剂, 并作为荧光探针应用于流式细胞术、荧光免疫和荧光显微镜5。
本协议着重于蓝藻Synechocystis模型中藻胆蛋白的定量测定。蓝藻是最早的氧光合自养;他们已经形成了地球的生物圈超过24亿年6。它们在氮、碳、氧和其他元素的全球生物地球化学循环中起着至关重要的作用。在蓝藻中, 单细胞菌株Synechocystis获得了独特的地位, 因为它是第一个蓝藻与整个基因组序列7,8, 它是自然转化的外源 DNA9, 并它执行稳定和相对地快速的成长10,11。在Synechocystis, 核心天线组件, 复方阿司匹林, 是与整体膜蛋白, 并附上藻位于囊体膜外围。
在该协议中比较了蛋白提取和定量的几种方法。最后提取程序成功地应用于Synechocystis, 以及其他蓝藻菌株, 包括Cyanothece sp, Synechococcuselongatus,螺旋藻sp,节旋藻sp和念珠菌, 也成功地应用于红藻类紫球藻藻。因此, 本协议中开发的方法可以被认为是蛋白提取的通用方法。尽管一些经过测试的提取方法导致了总蛋白产量的提高, 这里描述的萃取过程提供了最高的蛋白产量连同叶绿素的最低含量的残留在蛋白提取物。降低叶绿素a的含量是正确藻和复方阿司匹林分光光度定量的关键。
不同藻类和蓝藻物种的蛋白吸收光谱在12、13、14、15、16、17 、甚至在18种单一蓝藻属的几种菌株中。因此, 用于测定Synechocystis中藻和复方阿司匹林的特定波长和吸收系数一般不适用于其他菌株。此外, Synechocystis不包含藻红蛋白和 phycoerythrocyanin, 可以在其他藻类和蓝藻中发现。为了确定非Synechocystis菌株中的藻胆蛋白, 建议分别对各菌株的分光光度方程进行评价。
虽然该议定书包含两个较长的步骤 (夜间冷冻干燥的细胞颗粒和1小时蛋白质提取), 总的劳动时间为藻胆蛋白量化不超过2小时。

<table><tbody><tr><td>Synechocystis sp. PCC 6803</td><td>Institut Pasteur, Paris, France</td><td>6803</td><td>Cyanobacterium strain</td></tr><tr><td>Roti-CELL PBS</td><td>Carl Roth GmbH + Co. KG, Karlsruhe, Germany</td><td>9143.1</td><td>Phosphate-Buffered Saline (PBS) solution, pH 7.4</td></tr><tr><td>Eppendorf safe-lock tubes&amp;nbsp;</td><td>Eppendorf, Hamburk, Germany</td><td>30120086</td><td>Safe-lock tubes 1.5 ml</td></tr><tr><td>VWR 80-Place Storage System</td><td>VWR International, Radnor, Pennsylvania, USA</td><td>30128-282</td><td>Holder for safe-lock tubes&amp;nbsp;</td></tr><tr><td>RAININ 100 &amp;micro;l -1000 &amp;micro;l&amp;nbsp;</td><td>Mettler-Toledo, Columbus, Ohio, USA</td><td>17014382</td><td>Pipette</td></tr><tr><td>GP-LTS-A-1000&amp;micro;L-/F-768/8</td><td>Mettler-Toledo, Columbus, Ohio, USA</td><td>30389272</td><td>Pipette tips</td></tr><tr><td>Rotina 420R</td><td>Hettich, Kirchlengern, Germany</td><td>4701</td><td>Refrigerated centrifuge for 1.5 ml safe-lock tubes and 15 ml conical centrifuge tubes</td></tr><tr><td>LCexv 4010</td><td>Liebherr, Bulle, Switzerland</td><td>9005382197172</td><td>Refrigerator and freezer -20&amp;nbsp;&amp;deg;C</td></tr><tr><td>Revco ExF -86&amp;deg;C Upright Ultra-Low Temperature Freezer</td><td>Thermo Fisher Scientific, Waltham, Massachusetts, USA</td><td>EXF24086V&amp;nbsp;</td><td>Freezer -80&amp;nbsp;&amp;deg;C</td></tr><tr><td>CoolSafe</td><td>LaboGene, Liller&amp;oslash;d, Denmark</td><td>7.001.000.615</td><td>Freeze dryer&amp;nbsp;</td></tr><tr><td>UV-2600</td><td>Shimadzu, Kyoto, Japan</td><td>UV-2600</td><td>Spectrophotometer&amp;nbsp;</td></tr><tr><td>Hellma absorption cuvettes, semi Micro</td><td>Sigma-Aldrich, St. Louis, Missouri, USA</td><td>Z600288&amp;nbsp;</td><td>VIS/UV-VIS semi-micro cuvettes 0.75-1.5 ml, spectral range 200-2500 nm&amp;nbsp;</td></tr><tr><td>Silamat S6</td><td>Ivoclar Vivadent, Schaan, Liechtenstein</td><td>602286WU</td><td>Homogenizer&amp;nbsp;</td></tr><tr><td>Solid-glass beads</td><td>Sigma-Aldrich, St. Louis, Missouri, USA</td><td>Z273627</td><td>Glass bead of the diameter 2&amp;nbsp;mm</td></tr><tr><td>CPA225D-0CE</td><td>Sartorius AG, G&amp;ouml;ttingen, Germany</td><td>SECURA225D-1OBR</td><td>Analytical balances</td></tr><tr><td>C-Phycocyanin from Spirulina sp.&amp;nbsp;</td><td>Sigma-Aldrich, St. Louis, Missouri, USA</td><td>P2172</td><td>Phycocyanin standard</td></tr><tr><td>Allophycocyanin</td><td>Sigma-Aldrich, St. Louis, Missouri, USA</td><td>A7472</td><td>Allophycocyanin standard</td></tr><tr><td>Bicinchoninic Acid Kit&amp;nbsp;</td><td>Sigma-Aldrich, St. Louis, Missouri, USA</td><td>BCA1, B9643</td><td>Complete kit for total proteins determination</td></tr><tr><td>AlgaeTron&amp;nbsp;</td><td>Photon System Instruments Ltd., Dr&amp;aacute;sov, Czech Republic</td><td>AG 130-ECO&amp;nbsp;</td><td>Cultivation chamber for E. flasks, with controllable light and atmosphere</td></tr><tr><td>Photobioreactor</td><td>Photon System Instruments Ltd., Dr&amp;aacute;sov, Czech Republic</td><td>FMT-150</td><td>Cultivation equipment for cyanobacteria and algae with completely controllable environment</td></tr><tr><td>Cellometer&amp;nbsp;</td><td>Nexcelom Bioscience, Lawrence, Massachusetts, USA</td><td>Auto M10</td><td>Cell counter</td></tr><tr><td>Corning 15 mL centrifuge tubes</td><td>Sigma-Aldrich, St. Louis, Missouri, USA</td><td>CLS430791&amp;nbsp;</td><td>15 ml Centrifuge tube for dry weigth sampling</td></tr><tr><td>Herasafe KS</td><td>Thermo Fisher Scientific, Waltham, Massachusetts, USA</td><td>51024579</td><td>Laminar flow hood</td></tr></tbody></table>

spectrophotometric determination of phycobiliprotein content in cyanobacterium synechocystis

这是一个简单的协议, 以定量测定蛋白的内容在模型蓝藻Synechocystis。藻胆蛋白是 phycobilisomes 中最重要的成分, 是蓝藻中主要的光捕获触角和几种藻类群。Synechocystis的 phycobilisomes 包含两个藻胆蛋白: 藻和复方阿司匹林。该协议描述了一种简单、高效、可靠的方法, 用于定量测定该模型蓝藻中的藻和复方阿司匹林。比较了几种蛋白提取方法和分光光度法。本协议中所述的萃取过程也成功地应用于其他蓝藻菌株, 如Cyanothece 、 Synechococcuselongatus、螺旋藻、节旋藻sp、和念珠菌, 以及红藻类紫球藻藻。然而, 不同分类群的特定藻胆蛋白的消光系数可能不同, 因此建议分别验证每一个单株的分光光度法。该议定书需要很少的时间, 可以在任何标准生命科学实验室进行, 因为它只需要标准设备。

对于最初的方法测试, Synechocystis被培养了作为批培养在锥形烧瓶在 BG11 耕种媒介20 (补充与17毫米 HEPES) 在25°c, 在一个温暖的白色光之下强度的50µmol (光子)/(m2·s) 和与 1% CO2在培养的大气。在养殖过程中, 将这些培养物取样到安全锁管和离心 (实验室温度为5分钟 1.5万克), 将清液丢弃, 并将样品储存在-80 摄氏度, 冷冻干燥作为制?...

Watch this Scientific Journal Video about Spectrophotometric Determination of Phycobiliprotein Content in Cyanobacterium Synechocystis at JoVE.com

Spectrophotometric Determination of Phycobiliprotein Content in Cyanobacterium Synechocystis

在这里, 我们提出了一个协议, 以定量地确定蛋白的内容在蓝藻Synechocystis使用分光光度法。提取过程也成功地应用于其他蓝藻和藻类菌株;然而, 由于颜料吸收谱的变化, 有必要分别测试每种菌株的分光光度方程。

分光光度法测定蓝藻Synechocystis中的蛋白含量

This is a simple protocol for the quantitative determination of phycobiliprotein content in the model cyanobacterium Synechocystis. Phycobiliproteins are ...

spectrophotometric-determination-phycobiliprotein-content

Research

JoVE Journal

Biochemistry

Spectrophotometric Determination of Phycobiliprotein Content in Cyanobacterium Synechocystis

15.2K 次观看.  Czech Academy of Sciences. 在这里, 我们提出了一个协议, 以定量地确定蛋白的内容在蓝藻Synechocystis使用分光光度法。提取过程也成功地应用于其他蓝藻和藻类菌株;然而, 由于颜料吸收谱的变化, 有必要分别测试每种菌株的分光光度方程。

视频: Spectrophotometric Determination of Phycobiliprotein Content in Cyanobacterium Synechocystis

Determination of the Glycogen Content in Cyanobacteria

Cyanobacteria accumulate glycogen as a major intracellular carbon and energy storage&#160;during photosynthesis. Recent developments in research have highlighted complex mechanisms of glycogen metabolism, including the diel cycle of biosynthesis and catabolism, redox regulation, and the involvement of non-coding RNA. At the same time, efforts are being made to redirect carbon from glycogen to desirable products in genetically engineered cyanobacteria to enhance product yields. Several methods are used to determine the glycogen contents in cyanobacteria, with variable accuracies and technical complexities. Here, we provide a detailed protocol for the reliable determination of the glycogen content in cyanobacteria that can be performed in a standard life science laboratory. The protocol entails the selective precipitation of glycogen from the cell lysate and the enzymatic depolymerization of glycogen to generate glucose monomers, which are detected by a glucose oxidase-peroxidase (GOD-POD) enzyme coupled assay. The method has been applied to Synechocystis sp. PCC 6803 and Synechococcus sp. PCC 7002, two model cyanobacterial species that are widely used in metabolic engineering. Moreover, the method successfully showed differences in the glycogen contents between the wildtype and mutants defective in regulatory elements or glycogen biosynthetic genes.

Here, we present a reliable and easy assay to measure the glycogen content in cyanobacterial cells. The procedure entails precipitation, selectable depolymerization, and the detection of glucose residues. This method is suitable for both wildtype and genetically engineered strains and can facilitate the metabolic engineering of cyanobacteria.

测定蓝细菌中的糖原含量

Cyanobacteria accumulate glycogen as a major intracellular carbon and energy storage&#160;during photosynthesis. Recent developments in research have ...

科研

生物化学

Purification of Active Photosystem I-Light Harvesting Complex I from Plant Tissues

This method is used to isolate Photosystem I (PSI) together with the Light Harvesting Complex I (LHCI), its native antenna, from plants. PSI-LHCI is a large membrane protein complex coordinating hundreds of light harvesting and electron transport factors and is the most efficient light harvesting system found in nature. Photons absorbed by the four LHCA antenna proteins that make up LHCI are transferred through excitonic interaction to the PSI core reaction center and are used to facilitate light-driven charge separation across the thylakoid membrane, providing reducing power and energy for carbon fixation in photoautotrophic organisms. The high quantum efficiency of PSI makes this complex an excellent model to study light-driven energy transfer. In this protocol, plant tissue is mechanically homogenized, and the chloroplasts are separated from the bulk cellular debris by filtration and centrifugation. The isolated chloroplasts are then osmotically lysed, and the thylakoid membranes are recovered via centrifugation and solubilized using the detergent n-dodecyl-beta-maltoside. The solubilized material is loaded onto an anion exchange column to collect most of the chlorophyll-containing complexes. Larger complexes are precipitated from the solution, resuspended in a small volume, and loaded on sucrose gradients to separate the major chlorophyll-containing complexes. The resulting sucrose gradient fractions are characterized to identify the band of interest containing PSI-LHCI. This protocol is highly similar to the protocol used in the crystallization of plant PSI-LHCI with some simplifications and relies on methods developed over the years in the lab of Nathan Nelson.

This protocol describes the isolation of Photosystem I (PSI) - Light Harvesting Complex I (LHCI) from plant tissues. PSI along with PSII is responsible for the conversion of light to chemical energy in oxygenic photoautotrophs and has a quantum efficiency of ~1, making it a target for studying light-driven energy transfer.

This method is used to isolate Photosystem I (PSI) together with the Light Harvesting Complex I (LHCI), its native antenna, from plants. PSI-LHCI is a ...

<meta charset="utf8"></head><body>通过生长因子优化提高微藻中叶绿素的产生

Evaluation of the Effect of Growth Factors on Chlorophylls a and b Production from Microalgae

Microalgae contain two main groups of pigments: chlorophylls and carotenoids. Chlorophyll is a green pigment that absorbs light energy and transforms it into chemical energy to facilitate the synthesis of organic compounds. This pigment serves as a valuable primary source for biotechnological input products in the food, pharmaceutical, and cosmetic industries due to its high antioxidant properties and coloring capabilities. The objective of this research was to evaluate the effect of growth factors (CO2 concentration, light color, and light intensity) through a Taguchi L4 experimental design on cell growth and the cellular content of chlorophyll a and b in Chlorella sorokiniana, followed by validation of the method using Haematococcus pluvialis microalgae as an additional study model. Cell growth was quantified using the optical density spectrophotometric technique at a wavelength of 550 nm. For the quantification of chlorophylls, a cell extract was obtained using a 90% pure acetone solution, and subsequently, the concentrations of chlorophylls a and b were quantified using spectrophotometric techniques at wavelengths of 647 nm and 664 nm, according to the method described by Jeffrey and Humphrey. The experimental results indicated that controlling conditions of low CO2 addition, purple light, and low light intensity increases both cell growth and the concentration of chlorophylls a and b within the cells. The implementation of this chlorophyll quantification method allows for quick, simple, and precise determination of chlorophyll content, as the wavelengths used are at the absorbance peaks of both types of chlorophylls, making this technique easily reproducible for any microalgae under study.

<meta charset="utf8"></head><body>作者聚焦:优化生长因子以生产生物技术相关的次生代谢物

The present study highlights the advantages of employing the method developed by Jeffrey and Humphrey for extracting and quantifying fat-soluble pigments from microalgae. This method serves as a valuable tool for assessing the influence of growth factors on chlorophyll production and cellular content in these organisms.

生长因子对微藻叶绿素 a 和 b 生产的影响评价

Microalgae contain two main groups of pigments: chlorophylls and carotenoids. Chlorophyll is a green pigment that absorbs light energy and transforms it ...

Photobleaching Enables Super-resolution Imaging of the FtsZ Ring in the Cyanobacterium Prochlorococcus

Super-resolution microscopy has been widely used to study protein interactions and subcellular structures in many organisms. In photosynthetic organisms, however, the lateral resolution of super-resolution imaging is only ~100 nm. The low resolution is mainly due to the high autofluorescence background of photosynthetic cells caused by high-intensity lasers that are required for super-resolution imaging, such as stochastic optical reconstruction microscopy (STORM). Here, we describe a photobleaching-assisted STORM method which was developed recently for imaging the marine picocyanobacterium Prochlorococcus. After photobleaching, the autofluorescence of Prochlorococcus is effectively reduced so that STORM can be performed with a lateral resolution of ~10 nm. Using this method, we acquire the in vivo three-dimensional (3-D) organization of the FtsZ protein and characterize four different FtsZ ring morphologies during the cell cycle of Prochlorococcus. The method we describe here might be adopted for the super-resolution imaging of other photosynthetic organisms.

Photobleaching Enables Super-resolution Imaging of the FtsZ Ring in the Cyanobacterium Prochlorococcus

Here we describe a photobleaching method to reduce the autofluorescence of cyanobacteria. After photobleaching, stochastic optical reconstruction microscopy is used to obtain three-dimensional super-resolution images of the cyanobacterial FtsZ ring.

光漂白可实现前列腺素球菌中 ftsz 环的超分辨率成像

Super-resolution microscopy has been widely used to study protein interactions and subcellular structures in many organisms. In photosynthetic organisms, ...

免疫与感染

<meta charset="utf8"></head><body>与真菌伴侣共培养蔗糖分泌蓝细菌的方案

Assembly and Quantification of Co-Cultures Combining Heterotrophic Yeast with Phototrophic Sugar-Secreting Cyanobacteria

With the increasing demand for sustainable biotechnologies, mixed consortia containing a phototrophic microbe and heterotrophic partner species are being explored as a method for solar-driven bioproduction. One approach involves the use of CO2-fixing cyanobacteria that secrete organic carbon to support the metabolism of a co-cultivated heterotroph, which in turn transforms the carbon into higher-value goods or services. In this protocol, a technical description to assist the experimentalist in the establishment of a co-culture combining a sucrose-secreting cyanobacterial strain with a fungal partner(s), as represented by model yeast species, is provided. The protocol describes the key prerequisites for co-culture establishment: Defining the media composition, monitoring the growth characteristics of individual partners, and the analysis of mixed cultures with multiple species combined in the same growth vessel. Basic laboratory techniques for co-culture monitoring, including microscopy, cell counter, and single-cell flow cytometry, are summarized, and examples of nonproprietary software to use for data analysis of raw flow cytometry standard (FCS) files in line with FAIR (Findable, Accessible, Interoperable, Reusable) principles are provided. Finally, commentary on the bottlenecks and pitfalls frequently encountered when attempting to establish a co-culture with sugar-secreting cyanobacteria and a novel heterotrophic partner is included. This protocol provides a resource for researchers attempting to establish a new pair of co-cultured microbes that includes a cyanobacterium and a heterotrophic microbe.

<meta charset="utf8"></head><body>异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

This protocol provides a comprehensive guideline for the setup and quantitative monitoring of co-cultures, including photoautotrophic sugar-secreting cyanobacteria and heterotrophic yeasts.

异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

With the increasing demand for sustainable biotechnologies, mixed consortia containing a phototrophic microbe and heterotrophic partner species are being ...

生物工程

Measuring Photophysiology of Attached Stage of Colacium sp. by a Cuvette-Type Fast Repetition Rate Fluorometer

Fast repetition rate fluorometer (FRRf) is a beneficial method for measuring photosystem II (PSII) photophysiology and primary productivity. Although FRRf can measure PSII absorption cross-section (&#963;PSII), maximum photochemical efficiency (Fv/Fm), effective photochemical efficiency (Fq&#8242;/Fm&#8242;), and non-photochemical quenching (NPQNSV) for various eukaryotic algae and cyanobacteria, almost all FRRf studies to date have focused on phytoplankton. Here, the protocol describes how to measure PSII photophysiology of an epizoic alga Colacium sp.&#160;Ehrenberg 1834 (Euglenophyta), in its attached stage (attached to zooplankton), using cuvette-type FRRf. First, we estimated the effects of substrate zooplankton (Scapholeberis mucronata O.F. M&#252;ller 1776, Cladocera, Daphniidae) on baseline fluorescence and &#963;PSII, Fv/Fm, Fq&#8242;/Fm&#8242;, and NPQNSV of planktonic Colacium sp. To validate this methodology, we recorded photophysiology measurements of attached Colacium sp. on S. mucronata and compared these results with its planktonic stage. Representative results showed how the protocol could determine the effects of calcium (Ca) and manganese (Mn) on Colacium sp. photophysiology and identify the various effects of Mn enrichment between attached and planktonic stages. Finally, we discuss the adaptability of this protocol to other periphytic algae.

Measuring Photophysiology of Attached Stage of Colacium sp. by a Cuvette-Type Fast Repetition Rate Fluorometer

Fast repetition rate fluorometer (FRRf) is a beneficial method for measuring photosystem II photophysiology and primary productivity. Here we describe a protocol to measure PSII photophysiology of epizoic alga, Colacium sp. on substrate zooplankton using cuvette-type FRRf.

使用比色皿型快速重复率荧光计测量 Colacium sp.附着阶段的光生理学

Fast repetition rate fluorometer (FRRf) is a beneficial method for measuring photosystem II (PSII) photophysiology and primary productivity. Although FRRf ...

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