理化因子对集胞藻PCC6803溶血素溶血活性的影响

研究了保存温度以及添加稳定剂、氧化剂、还原剂、螯合剂和金属离子对集胞藻(Synechocystis sp. PCC6803)野生型菌株产生的溶血素的溶血活性的影响.结果表明,该溶血素在提取初始仅具有较低的溶血活性,但在提取3～6 d后,溶血活性急剧升高,之后随保存时间延长逐渐降低.-20 ℃低温保存有利于该溶血素活性的维持.添加半胱氨酸盐酸盐、牛血清白蛋白和血红蛋白都能提高其溶血活性,可以作为该溶血素的稳定剂;氧化剂H₂O₂能显著提高其溶血活性,而还原剂β-巯基乙醇、二硫苏糖醇显著抑制其溶血活性,说明该溶血素为非巯基依赖型溶血素.金属离子Ca2+,Co2+,Fe3+,Ni2+,Cu2+,Mn2+和Mg2+均会抑制其溶血活性,仅Zn2+能增加其溶血活性,金属螯合剂EDTA也能增强其溶血活性.      

Construction and application of the Synechocystis sp. PCC6803-ftnA in microbial contamination control in a coupled cultivation and wastewater treatment

Inspired by iron fertilization experiments in HNLC (high-nitrate, low-chlorophyll) sea areas, we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based on the genome sequence and natural transformation system of Synechocystis sp. PCC6803, ftnA (encoding ferritin) was selected as our target gene and was cloned into wild-type Synechocystis sp. PCC6803. Tests at the molecular level confirmed the successful construction of the engineered Synechocystis sp. PCC6803-ftnA. After Fe^3 +-EDTA pulsing, the intracellular iron content of Synechocystis sp. PCC6803-ftnA was significantly enhanced, and the algae was used in the microbial contamination control system. In the coupled Synechocystis sp. PCC6803-ftnA production and municipal wastewater (MW, including Scenedesmus obliquus and Bacillus) treatment, Synechocystis sp. PCC6803-ftnA accounted for all of the microbial activity and significantly increased from 70% of the microbial community to 95%. These results revealed that while the stored iron in the Synechocystis sp. PCC6803-ftnA cells was used for growth and reproduction of this microalga in the MW, the growth of other microbes was inhibited because of the iron limitation, and these results provide a new method for microbial contamination control during a coupling process.     

化感链霉菌6803菌株液体发酵工艺优化的研究

土壤微生物链霉菌6803菌株被证明对高等植物具有化感作用。采用单因子实验方法,研究液体发酵碳源、氮源、无机盐、发酵温度、发酵液初始pH值、摇床转速、发酵时间等对链霉菌6803菌株菌丝体产量及发酵液化感作用的影响,用均匀试验设计法优化了发酵工艺条件。结果表明：链霉菌6803菌株生长和产生化感作用的最佳发酵条件为：p（淀粉）=26．67g·L-1,p（蔗糖）=25．15g·L-1,p（NH4C1）=0．30g．L-1,黄豆饼粉浸液8．88％,所（NI-h）H2P04]=O．18g·L-1,p（FeS04．7H20）=O．002g·L-1,p（NaCl）=0．75g·L-1,pH值7．6,装量系数O．16,接种量3％,温度36℃,转速200r·min-1,发酵144h。本研究为利用微生物次生代谢产物作为天然除草剂奠定了基础。