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Methyl α-d-glucopyranoside enhances the enzymatic activity of recombinant β-galactosidase inclusion bodies in the araBAD promoter system of Escherichia coli

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

In this study, we utilized a catabolite repressor to improve the enzymatic activity of recombinant β-galactosidase inclusion bodies (IBs) produced in Escherichia coli under the araBAD promoter system. Specifically, we employed methyl α-d-glucopyranoside (α-MG) to lower the transcription rate of the β-galactosidase structural gene. In deepwell microtiter plate and lab-scale fermentor culture systems, we demonstrated that the addition of α-MG after induction improved the specific β-galactosidase production, even though β-galactosidase was still produced as an IB. Particularly, the addition of 0.0025% α-MG led to the most significant increase in the specific activity of the β-galactosidase. Interestingly, the β-galactosidase IBs obtained in the presence of 0.0025% α-MG were more loosely packed, as determined by IB solubilization in guanidine hydrochloride solution. We propose that the reduced gene transcription rate was responsible for the increased specific β-galactosidase activity and the loose packing that characterized the IBs produced in the presence of α-MG. This principle could be applied throughout the enzyme bioprocessing industry in order to enhance the activity of aggregate-prone enzymes within IBs.

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Acknowledgments

This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund) (KRF-2006-331-D00174). The author thanks Mr. Sung Hun Kim for his technical assistance.

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Authors and Affiliations

  1. Division of Food and Biotechnology, Chungju National University, Jeungpyung-Gun, Chungbuk, 368-701, South Korea

    Kyung-Hwan Jung, Ji-Hyeon Yeon & Sung-Kwon Moon

  2. BK21-NURI Committee, Korea Research Foundation, Yangjae-Dong, Seocho-Gu, Seoul, 137-943, Korea

    Joon Ho Choi

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  1. Kyung-Hwan Jung
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  2. Ji-Hyeon Yeon
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Correspondence to Kyung-Hwan Jung.

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Jung, KH., Yeon, JH., Moon, SK. et al. Methyl α-d-glucopyranoside enhances the enzymatic activity of recombinant β-galactosidase inclusion bodies in the araBAD promoter system of Escherichia coli . J Ind Microbiol Biotechnol 35, 695–701 (2008). https://doi.org/10.1007/s10295-008-0329-6

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  • DOI: https://doi.org/10.1007/s10295-008-0329-6

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