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.
【背景】大肠杆菌(Escherichia coli)由于生长性能优良、遗传背景清楚、遗传操作手段成熟,是合成β-法尼烯的合适生产菌,但其合成β-法尼烯的产量目前仍不能满足工业化生产的需求。【目的】通过诱变筛选技术选育β-法尼烯高产突变株。【方法】采用常压室温等离子体(atmosphericand room temperature plasma,ARTP)诱变技术和紫外线照射对出发菌株大肠杆菌EC-16进行复合诱变,并以异戊烯焦磷酸耐受性为选择压力进行平板初筛,之后进行摇瓶复筛,最后进行发酵罐验证。通过连续多代培养筛选到的高产突变菌株,观察其遗传稳定性。【结果】经复合诱变选育筛选出一株β-法尼烯高产突变株E.coliHVK-9,其产量高达22.1g/L,相比出发菌株提高了168.74%。【结论】采用ARTP-紫外复合诱变,再结合异戊烯焦磷酸抗性筛选的集成方法,使得诱变菌株的正突变率大大提高,可以有效地提高诱变菌株的β-法尼烯产量。突变株HVK-9作为工业化发酵生产菌种具有较好的遗传稳定性,为β-法尼烯的工业化生产和应用奠定了良好的基础。
