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Effects of feeding and induction strategy on the production of Bm R1 antigen in recombinant E. coli
Author(s) -
Norsyahida A.,
Rahmah N.,
Ahmad R.M.Y.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02694.x
Subject(s) - recombinant dna , antigen , fermentation , bioreactor , lac operon , biology , bacteria , yield (engineering) , microbiology and biotechnology , food science , biochemistry , immunology , botany , genetics , materials science , gene , metallurgy
Aim: To investigate the effects of feeding and induction strategies on the production of Bm R1 recombinant antigen. Methods and Results: Fed‐batch fermentation was studied with respect to the specific growth rate and mode of induction to assess the growth potential of the bacteria in a bioreactor and to produce high yield of Bm R1 recombinant antigen. Cells were grown at a controlled specific growth rate (μ set ) during pre‐induction, followed by constant feeding postinduction. The highest biomass (24·3 g l −1 ) was obtained during fed‐batch process operated at μ set of 0·15 h −1 , whereby lower μ set (0·075 h −1 ) gave the highest protein production (9·82 mg l −1 ). The yield of Bm R1 was increased by 1·2‐fold upon induction with 1 mmol l −1 IPTG (isopropyl‐β‐ d ‐thiogalactoside) compared to using 5 mmol l −1 and showed a further 3·5‐fold increase when the culture was induced twice at the late log phase. Conclusions: Combination of feeding at a lower μ set and twice induction with 1 mmol l −1 IPTG yielded the best result of all variables tested, promising an improved method for Bm R1 production . Significance and Impact of the Study: This method can be used to increase the production scale of the Bm R1 recombinant antigen to meet the increasing demand for Brugia Rapid ™ , a commercial diagnostic test for detection of brugian filariasis.