Bkt 基因和 crtR-B 基因在集胞藻 PCC 6803 中的重组表达及功能分析
Recombinant Expression and Functional Analysis of bkt Gene and crtR-B Gene in Synechocystis sp. PCC 6803
-
摘要: 将雨生红球藻中的 β-胡萝卜素酮化酶基因(bkt)和 β-胡萝卜素羟化酶基因(crtR-B)经密码子优化后, 通过自然转化法分别转入集胞藻 PCC 6803 基因组中。高效液相色谱分析显示：转入 bkt 基因的细胞产生角黄质的同时, 海胆酮含量减少；转入 crtR-B 基因的细胞产生金盏花黄质的同时, 玉米黄素含量减少。实验结果表明, 外源的 β-胡萝卜素酮化酶基因将海胆酮转化为角黄质, 外源的 β-胡萝卜素羟化酶基因将玉米黄素转化为金盏花黄质。该文利用代谢工程策略, 在集胞藻 PCC 6803 中构建类胡萝卜素生物合成途径, 为通过代谢工程在集胞藻 PCC 6803 中生产虾青素奠定了基础。Abstract: The β-carotene ketolase gene (bkt) and β-carotene hydroxylase gene (crtR-B) from Haematococcus pluvialis were codon-optimized and transferred to Synechocystis sp. PCC 6803 genome by natural transformation method. High performance liquid chromatography analysis showed that cells transfected with bkt gene produced canthaxanthin, while echinone decreased; the cells with crtR-B gene produced adonixanthin, while zeaxanthin was reduced. The results showed that the exogenous β-carotene ketolase converted echinone to canthaxanthin and the exogenous β-carotene hydroxylase converted zeaxanthin into adonixanthin. In this paper, the pathway of astaxanthin biosynthesis in Synechocystis sp. PCC 6803 was constructed by metabolic engineering strategy, which laid a foundation for astaxanthin production in Synechocystis sp. PCC 6803 with metabolic engineering.
下载: