Abstract
The transglutaminase (TGase) gene of Streptomyces fradiae was cloned. It had an ORF of 1242 bp, encoding a presumed prepro-region of 82 amino acids and a mature TGase of 331 amino acids. Enhanced expression of the TGase was achieved by introducing another copy of TGase gene into the original host genome which was driven by the strong constitutive promoter, “ermE up”, and shown to be expressed at the mRNA and protein levels. TGase activity in the recombinant strain (3.2 U/ml) was improved 1.3-fold when compared to that normally expressed in the original strain (2.4 U/ml). The specific enzyme activity in the recombinant strain (3.8 U/mg) was double that of the original strain (1.9 U/mg).
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Folk JE, Cole PW (1966) Mechanism of action of guinea pig liver transglutaminase. I. Purification and properties of the enzyme: identification of a functional cysteine essential for activity. J Biol Chem 241: 5518–5525
Folk JE (1980) Transglutaminases. Annu Rev Biochem 49:517–531
Date M, Yokoyama K, Umezawa Y, Matsui H, Kikuchi Y (2004) High level expression of Streptomyces mobaraensis transglutaminase in Corynebacterium glutamicum using a chimeric pro-region from Streptomyces cinnamoneus transglutaminase. J Biotechnol 110: 219–226
Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces Genetics. The John Innes Foundation, Norwich, UK
Kikuchi Y, Date M, Yokoyama K, Umezawa Y, Matsui H (2003) Secretion of active-form Streptoverticillium mobaraense transglutaminase by Corynebacterium glutamicum: processing of the pro-transglutaminase by a cosecreted subtilisin-Like protease from Streptomyces albogriseolus. Appl Environ Microbiol 69:358–366
Pasternack R, Dorsch S, Otterbaeh JT, Robenek IR, Wolf␣S, Fuehsbauer HL (1998) Bacterial pro-transglutaminase from Streptoverticillium mobaraense–purification, characterization and sequence of the zymogen. Eur J␣Biochem 257:570–576
Taguchi S, Arakawa K, Yokoyama K, Takehana S, Takagi H, Momose H (2002) Overexpression and purification of microbial pro-transglutaminase from Streptomyces cinnamoneum and in vitro processing by Streptomyces albogriseolus proteases. J Biosci Bioeng 94:478–481
von Mulert U, Luzhetskyy A, Hofmann C, Mayer A, Bechthold A (2004) Expression of the landomycin biosynthetic gene cluster in a PKS mutant of Streptomyces fradiae is dependent on the coexpression of a putative transcriptional activator gene. FEMS Microbiol Lett 230:91–97
Washizu K, Ando K, Koikeda S, Hirose S, Matsuura A, Takagi H, Motoki M, Takeuchi K (1994) Molecular cloning of the gene for microbial transglutaminase from Streptoverticillium and its expression in Streptomyces lividans. Biosci Biotechnol Biochem 58:82–87
Yokoyama K, Nakamura N, Seguro K, Kubota K (2000) Overproduction of microbial transglutaminase in Escherichia coli, in vitro refolding, and characterization of the refolded form. Biosci Biotechnol Biochem 64:1263–1270
Yokoyama K, Nio N, Kikuchi Y (2004) Properties and applications of microbial transglutaminase. Appl Microbiol Biotechnol 64:447–454
Yurimoto H, Yamane M, Kikuchi Y, Matsui H, Kato N, Sakai Y (2004) The pro-peptide of Streptomyces mobaraensis transglutaminase functions in cis and in trans to mediate efficient secretion of active enzyme from methylotrophic yeasts. Biosci Biotechnol Biochem 68:2058–2069
Zotzel J, Keller P, Fuchsbauer HL (2003a) Transglutaminase from Streptomyces mobaraensis is activated by an endogenous metalloprotease. Eur J Biochem 270:3214–3222
Zotzel J, Pasternack R, Pelzer C, Ziegert D, Mainusch M, Fuchsbauer HL (2003b) Activated transglutaminase from Streptomyces mobaraensis is processed by a tripeptidyl aminopeptidase in the final step. Eur J Biochem 270:4149–4155
Acknowledgements
This research was supported by Innovative Programs, the Chinese Academy of Science (KSCX2-SW-324). We thank Prof Keqian Yang, Dr Shenglan Wang (Institute of Microbiology, Chinese Academy of Sciences) and Prof Bin Hong (Chinese Academy of Medical Sciences) for providing plasmids and for helpful discussions during this work. We also thank Professor Deborah L. Segaloff (The University of Iowa, USA) for her help to edit the manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Liu, X., Yang, X., Xie, F. et al. Cloning of Transglutaminase Gene from Streptomyces fradiae and its Enhanced Expression in the Original Strain. Biotechnol Lett 28, 1319–1325 (2006). https://doi.org/10.1007/s10529-006-9094-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-006-9094-7