Deng M D, Coleman J R
Department of Botany, University of Toronto, Toronto, Ontario, M5S 3B2, Canada.
Appl Environ Microbiol. 1999 Feb;65(2):523-8. doi: 10.1128/AEM.65.2.523-528.1999.
Cyanobacteria are autotrophic prokaryotes which carry out oxygenic photosynthesis and accumulate glycogen as the major form of stored carbon. In this research, we introduced new genes into a cyanobacterium in order to create a novel pathway for fixed carbon utilization which results in the synthesis of ethanol. The coding sequences of pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adh) from the bacterium Zymomonas mobilis were cloned into the shuttle vector pCB4 and then used to transform the cyanobacterium Synechococcus sp. strain PCC 7942. Under control of the promoter from the rbcLS operon encoding the cyanobacterial ribulose-1, 5-bisphosphate carboxylase/oxygenase, the pdc and adh genes were expressed at high levels, as demonstrated by Western blotting and enzyme activity analyses. The transformed cyanobacterium synthesized ethanol, which diffused from the cells into the culture medium. As cyanobacteria have simple growth requirements and use light, CO2, and inorganic elements efficiently, production of ethanol by cyanobacteria is a potential system for bioconversion of solar energy and CO2 into a valuable resource.
蓝细菌是进行产氧光合作用并将糖原作为主要碳储存形式积累的自养原核生物。在本研究中,我们将新基因导入蓝细菌,以创建一条固定碳利用的新途径,从而合成乙醇。运动发酵单胞菌的丙酮酸脱羧酶(pdc)和乙醇脱氢酶II(adh)的编码序列被克隆到穿梭载体pCB4中,然后用于转化蓝细菌聚球藻属PCC 7942菌株。在编码蓝细菌核酮糖-1,5-二磷酸羧化酶/加氧酶的rbcLS操纵子启动子的控制下,pdc和adh基因高水平表达,蛋白质免疫印迹和酶活性分析证明了这一点。转化后的蓝细菌合成了乙醇,乙醇从细胞扩散到培养基中。由于蓝细菌生长需求简单且能高效利用光、二氧化碳和无机元素,利用蓝细菌生产乙醇是将太阳能和二氧化碳生物转化为有价值资源的潜在系统。