Yildirim Selcuk, Konrad Daniel, Calvez Ségolène, Drider Djamel, Prévost Hervé, Lacroix Christophe
Laboratory of Food Biotechnology, Institute of Food Science and Nutrition, ETH (Swiss Federal Institute of Technology) Zurich, LFV C20 Schmelzbergstrasse 7, CH-8092, Zurich, Switzerland.
Appl Microbiol Biotechnol. 2007 Dec;77(3):525-31. doi: 10.1007/s00253-007-1188-1. Epub 2007 Sep 19.
To increase the yield of heterologous production of the class II bacteriocin DvnRV41 with Escherichia coli Origami (DE3) (pLysS/pCR03), induction of bacteriocin gene expression was optimized by varying the inducer isopropyl beta-D-thiogalactopyranoside (IPTG) concentration (0-2 mM), and controlled batch and fed-batch cultures were tested on a 2-L scale. A concentration of 0.5 mM IPTG was found to be optimal for cell growth and bacteriocin production. Shake flask cultivation of E. coli Origami (DE3) (pLysS/pCR03) gave biomass and bacteriocin yields of 1.54 +/- 0.06 g cdw/l and 18 +/- 1 mg DvnRV41/l, respectively. Biomass (2.70 +/- 0.06 and 6.8 +/- 0.6 g cdw/l, respectively) and bacteriocin yields (30 and 74 mg DvnRV41 per liter, respectively) were both increased with batch and fed-batch compared to shake flask cultures. Bacteriocin yields reported in this study are among the highest published for other heterologous expression systems in shake flasks.
为提高用大肠杆菌Origami(DE3)(pLysS/pCR03)进行II类细菌素DvnRV41异源生产的产量,通过改变诱导剂异丙基-β-D-硫代半乳糖苷(IPTG)浓度(0-2 mM)优化细菌素基因表达的诱导,并在2-L规模上测试了分批补料和补料分批培养。发现0.5 mM IPTG的浓度最适合细胞生长和细菌素生产。大肠杆菌Origami(DE3)(pLysS/pCR03)的摇瓶培养分别得到1.54±0.06 g干重/L的生物量和18±1 mg DvnRV41/L的细菌素产量。与摇瓶培养相比,分批补料和补料分批培养的生物量(分别为2.70±0.06和6.8±0.6 g干重/L)和细菌素产量(分别为每升30和74 mg DvnRV41)均有所增加。本研究报道的细菌素产量是摇瓶中其他异源表达系统已发表的最高产量之一。