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High Density Fed-Batch Culture of Escherichia coli BL21/pET28b-glnA With DO Feed-back Control of Nutrient Feeding(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2010年02期
Page:
96-103
Research Field:
生命科学
Publishing date:

Info

Title:
High Density Fed-Batch Culture of Escherichia coli BL21/pET28b-glnA With DO Feed-back Control of Nutrient Feeding
Author(s):
Zhang Feng; Wang Qi; Li Guilan; Xu Jianhong; Hong Suli; Yue Ling; Feng Zhiyong; Yin Zhimin
School of Life Science,Nanjing Normal University,Nanjing 210046,China
Keywords:
G lutam ine synthetase;  recomb inan tE. co li;  fed -batch cu lture;  h igh cell dens ity cu lture
PACS:
TQ925
DOI:
-
Abstract:
To investigate the optim a l h igh cell density culture conditions of recomb inant BL21 /pET28b-g lnA to produce recom b inant G lutam ine Synthetase ( GS) of Bac illus Subtilis, based on the exper im enta l resu lts obtained in shake flask cu ltivation, the effects of the composition o f the cu lturem edium, the range o f pH, induce r concen tration and induced tim e on the g row th o f bac teria and the expression leve l o f GS we re ana lyzed. Then the bacter ia we re transferred to B IOTECH- 5BG( 5 L) DO feedback fed ba tch cu lture system. By keep ing dissolved oxygen at 20% ~ 50%, pH at 7.0~ 7.5, cultu ring in Optim izedM9M edium and induc ing for 12 h in the presence o f 1 g /L lactose at 37℃ during fedbatch cu ltiva tion, we perform ed a three tim e repeated culture. Ce ll density of recomb inan t E. co li w as reached to 48.1 g /L under constant ( 0.10 h - 1 ) specific g row th rates. The express ion leve l o f recom binant prote in GS w as over 55% o f the to tal pro te in in E. coli. Specific ac tiv ity o f crude ex trac t enzyme w as 10.35 U /mg. A h igh expression and stab ilized cu lture process o f recomb inan t Glutam ine Synthe tase of Bac illus Subtilis has been established, wh ich founds the basis for further larg e sca le produc tion o f G lutam ine Synthe tase.

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Last Update: 2013-04-08