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Heterologous signal peptides-directing secretion of Streptomyces mobaraensis transglutaminase by Bacillus subtilis

Mu, D., Lu, J., Qiao, M., Kuipers, O. P., Zhu, J., Li, X., Yang, P., Zhao, Y., Luo, S., Wu, X., Jiang, S. & Zheng, Z., Jul-2018, In : Applied Microbiology and Biotechnology. 102, 13, p. 5533-5543 11 p.

Research output: Contribution to journalArticleAcademicpeer-review

  • Dongdong Mu
  • Jiaojiao Lu
  • Mingqiang Qiao
  • Oscar P Kuipers
  • Jing Zhu
  • Xingjiang Li
  • Peizhou Yang
  • Yanyan Zhao
  • Shuizhong Luo
  • Xuefeng Wu
  • Shaotong Jiang
  • Zhi Zheng

Microbial transglutaminase (MTG) from Streptomyces mobaraensis has been widely used for crosslinking proteins in order to acquire products with improved properties. To improve the yield and enable a facile and efficient purification process, recombinant vectors, harboring various heterologous signal peptide-encoding fragments fused to the mtg gene, were constructed in Escherichia coli and then expressed in Bacillus subtilis. Signal peptides of both WapA and AmyQ (SP wapA and SP amyQ ) were able to direct the secretion of pre-pro-MTG into the medium. A constitutive promoter (P hpaII ) was used for the expression of SP wapA -mtg, while an inducible promoter (P lac ) was used for SP amyQ -mtg. After purification from the supernatant of the culture by immobilized metal affinity chromatography and proteolysis by trypsin, 63.0 ± 0.6 mg/L mature MTG was released, demonstrated to have 29.6 ± 0.9 U/mg enzymatic activity and shown to crosslink soy protein properly. This is the first report on secretion of S. mobaraensis MTG from B. subtilis, with similar enzymatic activities and yields to that produced from Escherichia coli, but enabling a much easier purification process.

Original languageEnglish
Pages (from-to)5533-5543
Number of pages11
JournalApplied Microbiology and Biotechnology
Volume102
Issue number13
Early online date25-Apr-2018
Publication statusPublished - Jul-2018

    Keywords

  • Transglutaminase, Bacillus subtilis, Secretion, Signal peptides

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