Calcium carbonate (CaCO3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 (inoculated BG11). In this study, the features of calcium carbonate d...Calcium carbonate (CaCO3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 (inoculated BG11). In this study, the features of calcium carbonate deposition were investigated. Inoculated BGll in different calcium ion concentrations was used for the experimental group, while the BGll culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium (Ca), carbon (C), oxygen (O), phosphorus (P), iron (Fe), copper (Cu), zinc (Zn), and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BGll culture media. There may be more calcium- containing crystals inside and outside of these cells. These results shall be beneficial for understanding the formation mechanism of carbonate minerals.展开更多
【背景】蛋白酶能够降解细胞中错误折叠或是无功能的蛋白,Clp家族蛋白就是一类重要的蛋白酶复合物。Clp蛋白酶复合物的水解核心是ClpP,集胞藻PCC6803中存在4种不同的ClpP蛋白,分别为ClpP1-ClpP4。作为重要的蛋白水解复合物的功能组分,...【背景】蛋白酶能够降解细胞中错误折叠或是无功能的蛋白,Clp家族蛋白就是一类重要的蛋白酶复合物。Clp蛋白酶复合物的水解核心是ClpP,集胞藻PCC6803中存在4种不同的ClpP蛋白,分别为ClpP1-ClpP4。作为重要的蛋白水解复合物的功能组分,目前对集胞藻ClpP的研究十分有限,对其生理功能与调控底物的研究甚少。【目的】选择集胞藻为研究对象探究ClpP2蛋白的功能,鉴定其潜在底物,为集胞藻ClpP2作用机制提供实验支撑。【方法】构建集胞藻ClpP2突变株(ΔClpP2),进行其生长实验和光合生理功能研究。通过标记定量蛋白质组学技术(isobaric tag for relative absolute quantitation,iTRAQ)鉴定ClpP2调控的靶标蛋白,生物信息学分析底物蛋白参与的代谢通路,最后利用平行反应监测(parallel reaction monitoring,PRM)技术对部分定量数据进行验证。【结果】ΔClpP2可以在自然条件下光合自养生长至对数生长期,但高光或高温胁迫下则无法正常生长。相较于野生型,ΔClpP2有着显著降低的PSⅡ电子传递效率及PSⅠ环式电子传递活性。通过iTRAQ定量蛋白质组学手段,ΔClpP2相对于WT共鉴定到206个差异表达蛋白,其中131个上调、75个下调,为ClpP2蛋白酶提供了丰富的潜在底物库。基因本体论(gene ontology,GO)分析发现ClpP2主要参与各种物质的转运,其中ABC蛋白转运途径显著被富集。利用PRM技术对34个差异表达的蛋白进行了验证。【结论】ClpP2蛋白不是集胞藻生长所必需,但在高温或者高光胁迫下是必不可少的,其失活会降低集胞藻光合系统活性。ClpP2可能通过调控离子转运进而影响光合系统。ClpP2很可能与ClpX结合形成蛋白酶复合物。展开更多
In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature (LT) stress. Among the genes on the cDN...In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature (LT) stress. Among the genes on the cDNA microarrays, 899 LT-affected genes exhibited a 1.5-fold (or greater) difference in expression compared with the genes from normal unstressed Synechocystis sp. PCC6803. Of the differentially expressed genes, 353 were up-regulated and 246 were down-regulated. The results showed that genes involved in photosynthesis were activated at LT (10℃), including genes for photosystem I, photosystem II, photosynthetic electron transport, and cytochrome b6/f complex. Moreover, desg, one of four genes that encode the fatty acid desaturases, was also induced by LT. However, the LT conditions to some degree enhanced the transcription of some genes. In addition, LT (10℃) may reduce cellular motility by regulating the transcription of spkA (sll1575), a serine/threonine protein kinase. The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT, including pathways involved in photosynthesis and repair.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.40972043,41040018,41210104058,21176145,41372108,41302079)the Higher Educational Science and Technology Program of Shandong Province(No.J10LC15)+4 种基金the China Postdoctoral Science Foundation(No.2013M540560)the Program for Scientific Research Innovation Team in Colleges and Universities of Shandong Province,and SDUST Research Fund(No.2010KYTD103)the Open Project of Key Lab of Marine Bioactive Substance and Modern Analytical Technique,State Oceanic Administration,China(No.MBSMAT-2012-03)the Scientific and Technological Program of Qingdao(No.13-1-4-232-jch)the Domestic Visiting Scholar Program for Young Core Teachers in Shandong Universities,Shandong Province,China
文摘Calcium carbonate (CaCO3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 (inoculated BG11). In this study, the features of calcium carbonate deposition were investigated. Inoculated BGll in different calcium ion concentrations was used for the experimental group, while the BGll culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium (Ca), carbon (C), oxygen (O), phosphorus (P), iron (Fe), copper (Cu), zinc (Zn), and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BGll culture media. There may be more calcium- containing crystals inside and outside of these cells. These results shall be beneficial for understanding the formation mechanism of carbonate minerals.
文摘【背景】蛋白酶能够降解细胞中错误折叠或是无功能的蛋白,Clp家族蛋白就是一类重要的蛋白酶复合物。Clp蛋白酶复合物的水解核心是ClpP,集胞藻PCC6803中存在4种不同的ClpP蛋白,分别为ClpP1-ClpP4。作为重要的蛋白水解复合物的功能组分,目前对集胞藻ClpP的研究十分有限,对其生理功能与调控底物的研究甚少。【目的】选择集胞藻为研究对象探究ClpP2蛋白的功能,鉴定其潜在底物,为集胞藻ClpP2作用机制提供实验支撑。【方法】构建集胞藻ClpP2突变株(ΔClpP2),进行其生长实验和光合生理功能研究。通过标记定量蛋白质组学技术(isobaric tag for relative absolute quantitation,iTRAQ)鉴定ClpP2调控的靶标蛋白,生物信息学分析底物蛋白参与的代谢通路,最后利用平行反应监测(parallel reaction monitoring,PRM)技术对部分定量数据进行验证。【结果】ΔClpP2可以在自然条件下光合自养生长至对数生长期,但高光或高温胁迫下则无法正常生长。相较于野生型,ΔClpP2有着显著降低的PSⅡ电子传递效率及PSⅠ环式电子传递活性。通过iTRAQ定量蛋白质组学手段,ΔClpP2相对于WT共鉴定到206个差异表达蛋白,其中131个上调、75个下调,为ClpP2蛋白酶提供了丰富的潜在底物库。基因本体论(gene ontology,GO)分析发现ClpP2主要参与各种物质的转运,其中ABC蛋白转运途径显著被富集。利用PRM技术对34个差异表达的蛋白进行了验证。【结论】ClpP2蛋白不是集胞藻生长所必需,但在高温或者高光胁迫下是必不可少的,其失活会降低集胞藻光合系统活性。ClpP2可能通过调控离子转运进而影响光合系统。ClpP2很可能与ClpX结合形成蛋白酶复合物。
基金Supported by the National Natural Science Foundation of China(No.40876082)the International Innovation Partnership Program:Typical Environmental Process and Effects on Resources in Coastal Zone Area+3 种基金the Public Science and Technology Research Funds Projects of the Ocean(Nos.200905021,201205027)the Outstanding Young Scholars Fellowship of Shandong Province(Molecular Phycology,No.JQ200914)the Natural Science Foundation of Shandong Province(No.ZR2012DQ015)the Guangdong Province Comprehensive Strategic Cooperation Project of the Chinese Academy of Sciences(No.2011A090100040)
文摘In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature (LT) stress. Among the genes on the cDNA microarrays, 899 LT-affected genes exhibited a 1.5-fold (or greater) difference in expression compared with the genes from normal unstressed Synechocystis sp. PCC6803. Of the differentially expressed genes, 353 were up-regulated and 246 were down-regulated. The results showed that genes involved in photosynthesis were activated at LT (10℃), including genes for photosystem I, photosystem II, photosynthetic electron transport, and cytochrome b6/f complex. Moreover, desg, one of four genes that encode the fatty acid desaturases, was also induced by LT. However, the LT conditions to some degree enhanced the transcription of some genes. In addition, LT (10℃) may reduce cellular motility by regulating the transcription of spkA (sll1575), a serine/threonine protein kinase. The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT, including pathways involved in photosynthesis and repair.