基因工程菌高密度发酵液中碳源物质甘油的定量检测及其浓度的优化控制

比较了高碘酸氧化法，铜离子络合法、高压液相色谱法和甘油激酶法四种不同的方法定量测定重组工程菌ＹＫ５３７／ＰＳＢ－ＴＫ高密度发酵液中甘油的浓度，发现甘油激酶法可以排除发酵液中其他物质的干扰，精确测定甘油的含量。在此基础上对发酵培养中甘油的浓度进行了优化，结果表明，甘油浓度控制在较低的水平有利于菌体密度的提高。在５Ｌ自动发酵罐中，控制甘油浓度在５ｇ／Ｌ左右，经２０ｈ的培养，最终细菌密度达到１２０ＯＤ６

Determination and optimization of glycerol during the high density fermentation of genetic engineered bacteria

This article described the determination of glycerol contents in the culture broth during the high density fermentation of reconmbinant YK537/pSB TK with periodic test,cupric ion complex,HPLC,glycerokinase (GK) and all these methods were compared.Among them,enzymatic methods (GK) could accurately determine the glycerol contents avoiding the interference in the broth.Based on the analysis,glycerol concentration during fermentation was optimized.Results showed that low glycerol concentration was favorable to increasing cell density.Keeping glycerol concentration at 5 g/L,cell density reached 120 OD 600 (corresponding to 48 g DCW/L) after 20 h cultivation in a 5L autocontrolled fermentor,expressed recombinant tumor necroses factor was about 30% of total cellular protein.