封闭式光生物反应器集胞藻6803光自养和混合营养培养比较

采用封闭式光生物反应器进行了蓝藻基因工程常用宿主系统集胞藻Synechocystis sp. PCC 6803的混合营养培养,并与光自养培养进行了比较,在两种培养方式下,集胞藻6803的饱和光强基本相同,都为5000lx,当入射光强为5000lx,初始葡萄糖浓度为1．74g/L时,混合营养生长在葡萄糖消耗完（69．5h)时的藻细胞密度为1．36g/L,叶绿素浓度为20．08mg/L,能量得率为16．7％,分别为同期光自养生长的3．8倍,2．3倍和2．6倍,这表明封闭式光生物反应器混合营养培养方式在促进集胞藻6803生长和光合色素合成及提高培养过程能量得率等方面都有显著作用。     

集胞藻PCC6803基因ssl1690缺失突变株的构建

使用BLAST软件对聚球藻PCC7002的裂合酶基因cpcT进行了同源搜索分析,在集胞藻PCC6803中获取了相似程度达68%的同源基因ssl1690.为进一步探讨ssl1690功能,构建了同源缺失突变载体,将其转入蓝藻细胞中,筛选得到基因缺失突变株,并将基因缺失突变株培养于BG11液体培养基中,观察了其生长情况和表型变化.结果表明:基因缺失突变株与野生株相比,生长有所延迟,有漂白现象.通过分离和比较SDS-PAGE电泳突变株与野生株的藻胆体蛋白,发现缺失株存在藻胆体组分蛋白的缺失.故认为集胞藻PCC6803 ssl1690基因功能与光合作用有关,其所编码的蛋白对藻蓝蛋白正常的体内生物合成具有重要作用.     

绿色荧光蛋白标记S2P在集胞藻6803中的表达

为研究集胞藻6803中两个S2P同源蛋白酶Slr0643和Sll0862的功能,分别在两个蛋白酶的羧基端添加增强型绿色荧光蛋白(EGFP)标签,构建了两株转基因集胞藻psbA2::0643GFP-Cmr/△0643-Kmr和psbA2::0862GFP-Cmr/△0862-Kmr.通过逆转录聚合酶链式反应(RT-PCR)检测到egfp基因的转录表达,通过激光共聚焦显微镜检测到转基因集胞藻细胞发出的绿色荧光蛋白荧光,表明带EGFP标签的S2P融合蛋白在psbA2启动子调控下正确表达.     

集胞藻PCC6803基因sll0853的克隆、优化表达及其结构功能预测

从集胞藻(Synechocystis sp.)PCC6803提取总DNA,利用PCR扩增目的基因sll0853,构建重组T-0853克隆载体和pET-0853原核表达载体.为了提高sll0853在大肠杆菌E.coli BL21(DE3)中的表达量,通过改变诱导温度、诱导时间及诱导剂浓度等条件对表达量产生影响,以SDS-PAGE电泳分析证明sll0853基因蛋白表达的最佳条件.结果表明:目的蛋白在28℃、0.2mmol/L IPTG、诱导6h表达量分别达高峰.通过生物信息学软件预测基因sll0853可能具有裂合酶的功能.     

转小鼠金属硫蛋白基因集胞藻6803及其野生藻培养

通过摇瓶培养考察转小鼠金属硫蛋白基因集胞藻６８０３及其野生藻的光合自养培养过程中碳源浓度、氮源浓度和光照度对这两种藻生长的影响。野生藻的最适生长碳源浓度（０．０５－０．５ｇ／Ｌ）比转基因藻的最适生长碳源浓度（０．０２－０．０５ｇ／Ｌ）高。氮源浓度对转基因藻和野生藻的影响趋势相同。在０．３－４．５ｇ／Ｌ范围内,随着氮源浓度的增高,藻体生长的最大细胞浓度降低。转基因藻生长的最佳光照度为１５００ｌｘ,野     

镉对集胞藻PCC6803生长的影响

以集胞藻为研究材料,研究不同浓度的Cd2 处理对集胞藻生长的影响.研究发现:低浓度的Cd2 处理(0~0.1 m g/L),能够促进集胞藻的生长,表现为叶绿素、可溶性糖、蛋白质含量随着浓度的增大而增大;当Cd2 浓度进一步增加时,Cd2 开始抑制集胞藻的生长,表现为叶绿素、可溶性糖、蛋白质含量减小,Cd2 浓度达到1.0 m g/L则完全抑制集胞藻的生长,无任何产物累积.     

集胞藻胞外多糖的流变学性质和乳化活性

集胞藻在培养过程中向培养基中分泌大量胞外多糖,流变学研究表明这种胞外多糖呈现假塑性行为,粘度值接近黄原胶,当剪切速率在0.15 S-1时,两者粘度相等.集胞藻胞外多糖在0.1 mol/L氯化钠溶液中以及在30～70℃温度范围内粘度值具有一定的稳定性.集胞藻胞外多糖的粘弹谱类似于黄原胶.具有弱凝胶的特征.这种多糖具有良好稳定的乳化活性,对植物油、正十六烷、石蜡、二甲苯和2-甲基萘 正十六烷(1:1)都有乳化作用,而且在静置一个月后,乳液依然稳定.集胞藻胞外多糖可以作为增稠剂、稳定剂以及乳化剂在食品、化妆品工业等领域得到广泛应用.     

平板式光生物反应器在饵料微藻培养中的应用

对自主研发的平板式光生物反应器的主要性能参数进行了测定,并以纤细角毛藻和等鞭金藻为培养对象对平板反应器培养微藻的条件进行研究.结果表明,光照和通气速率是影响微藻在反应器中生长的主要因素,一定范围内高光照强度和高通气速率有利于微藻细胞的生长.通过与开放式培养系统对照培养表明,平板反应器能够有效提高培养密度.纤细角毛藻在培养8 d后,仍然能够保持缓慢的增殖状态,细胞生长趋于稳定期,培养末期的细胞密度达到5.01×108个/mL,高于开放系统培养的1.81×107个/mL.该平板反应器具有操作简单、容易放大、价格低廉等优势,适用于工业化生产.     

平板式光生物反应器的Parietochloris incisa超高密度培养

利用平板式光生物反应器对一种新分离的微藻Parietochlorisincisa进行了室外放大培养研究。在最适培养条件下 ,实现了对该微藻的超高密度培养 ,使单位培养体积和单位培养面积的细胞生物量分别达到了 7.35g/ (L·d)和 96 .12 g/ (m2 ·d)     

Mixotrophic Cultivation of Tetraselmis sp. - 1 in Airlift Photobioreactor

Tetraselmis sp. -1 is a new microalgae strain constructed by cell fusion technique. In this paper, the mixotrophic cultivation of Tetraselmis sp.-1 in airlift photobioreactor is investigated.Firstly, the paper calculates the light attenuation in the mixotrophic medium, and sets the light attenuation model. Secondly, it uses the same dissolved oxygen coefficient (Ka) of flask culture to select the aeration of bioreactor. Finally, it sets the growth kinetic model, production (chlorophyll - a and total lipid) kinetic models and substrate (glucose) consumption kinetic model of Tetraselmis sp. - 1 in airlift photobioreactor.     

Sodium Acetate Stimulates PHB Biosynthesis in Synechocystis sp. PCC 6803

IntroductionA large variety of bacteria are able to synthesizepolyhydroxyalkanoic acids (PHAs) and to depositlarge amounts of these polyesters as insolubleinclusions in cytoplasm[1 ] . PHAs have attractedthe interest of the chemical industry because oftheir biodegradable properties for applications invarious technical,medical,and pharmaceuticalapplications[2 ] .Poly-β-hydroxybutyrate(PHB) isatypical PHA that is produced by heterotrophicbacteria,such as Alcaligenes eutrophus andEscherichi…     

Two-Component Signal Transduction Systems in the Cyanobacterium Synechocystis sp. PCC 6803

Introduction Rapid adaptation to environmental challenges is vitally important for bacterial survival and growth. One way in which bacteria control their response to changing environmental conditions is through the mechanism of two-component signal transd…     

Role of two phosphohexomutase genes in glycogen synthesis in Synechocystis sp. PCC6803

Phosphohexomutases catalyze the interconversion between hexose-6-phosphate and hexose-l-phosphate and play important roles in polysaccharide synthesis. In Synechocystis sp. PCC 6803, sl10726 is predicted to encode PGM （phosphoglucomutase）, slr1334 is predicted to encode a PGM/PMM （phosphomannomutase） bifunction enzyme. In comparison to the wild type, a sllO726-null mutant showed 3.4% PGM activity but 45%-69% glycogen content. Down-regulation of slr1334, an essential gene, by using a copper regulated promoter further decreased the PGM activity in the sllO726：：Kmr PpetE-slr1334 double mutant to 0.3% of the wild type level. However, the glycogen content was not further decreased in parallel. In vitro, recombinant Sl10726 or S1r1334 showed predicted enzyme activities. Our results indicate that a relatively high level of glycogen can be maintained in Synechocystis mutants with low levels of PGM activity. The high PGM activity in the cyanobacterium may be required for turnover of glycogen or synthesis of other polysaccharides or oligosaccharides.     

Transient Effects of Salt Shock on the Photosynthetic Machinery in Synechocystis sp. PCC 6803

IntroductionSalt stress is one of the main detrimental factors inthe environment that limit the growth andproductivity of plants.Salt stress causessignificant decreases in photosynthetic activity,such as the electron transport[1,2 ] ,but themechanisms by which salt stress inhibitsphotosynthesis remain poorly understood[3] .　Cyanobacteria are prokaryotes that performoxygenic photosynthesis using a photosyntheticapparatus similar to that in the chloroplasts ofgreen algae and higher plants[4 ] .…