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A novel approach for improving the yield of Bacillus subtilis transglutaminase in heterologous strains

  • Biotechnology Methods
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The transglutaminase (BTG) from Bacillus subtilis is considered to be a new type of transglutaminase for the food industry. Given that the BTG gene only encodes a mature peptide, the expression of BTG in heterologous microbial hosts could affect their normal growth due to BTG’s typical transglutaminase activity which can catalyze cross-linking of proteins in the cells. Therefore, we developed a novel approach to suppress BTG activity and reduce the toxicity on microbial hosts, thus improving BTG yield. Genes encoding the respective regions of transglutaminase propeptide from seven species of Streptomyces were fused to the N-terminal of the BTG gene to produce fusion proteins. We found that all the fused propeptides could suppress BTG activity. Importantly, BTG activity could be completely restored after the removal of the propeptides by proteolytic cleavage. Of the seven propeptides tested, the propeptide proD from Streptomyces caniferus had the strongest suppressive effect on BTG activity (70 % of the activity suppressed). Moreover, fusion protein proD-BTG (containing proD) also exhibited the highest yield which was more than twofold of the expression level of BTG in an active form in Escherichia coli. Secretion expression of BTG and proD-BTG in Corynebacterium glutamicum further showed that our novel approach was suitable for the efficient BTG expression, thus providing a valuable platform for further optimization of large-scale BTG production.

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Acknowledgments

This work was supported by the National High-Tech Research and Development Plan (“863” Plan) (2011AA100905-4), the Tianjin Research Program of Application Foundation and Advanced Technology (14JCYBJC23800), the programme for Changjiang Scholars and Innovative Research Team in University (IRT1166), and the National Natural Science Fund (31101219) of China.

Conflict of interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled ‘A novel approach for improving the yield of Bacillus subtilis transglutaminase in heterologous strains’.

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Authors and Affiliations

  1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, National Engineering Laboratory for Industrial Enzymes, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin, People’s Republic of China

    Yihan Liu, Song Lin, Xiqing Zhang, Xiaoguang Liu, Jianling Wang & Fuping Lu

  2. No. 29, 13th Avenue, Tianjin Economic and Technological Development Area, 08 Postbox, Tianjin, 300457, People’s Republic of China

    Fuping Lu

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  1. Yihan Liu
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  2. Song Lin
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  3. Xiqing Zhang
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  4. Xiaoguang Liu
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Correspondence to Fuping Lu.

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Liu, Y., Lin, S., Zhang, X. et al. A novel approach for improving the yield of Bacillus subtilis transglutaminase in heterologous strains. J Ind Microbiol Biotechnol 41, 1227–1235 (2014). https://doi.org/10.1007/s10295-014-1468-6

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  • DOI: https://doi.org/10.1007/s10295-014-1468-6

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