Journal of Industrial Microbiology & Biotechnology

, Volume 35, Issue 9, pp 981–990 | Cite as

Characterization and large-scale production of recombinant Streptoverticillium platensis transglutaminase

  • Shie-Jea Lin
  • Yi-Fang Hsieh
  • Li-An Lai
  • Mei-Li Chao
  • Wen-Shen Chu
Original Paper

Abstract

Recombinant Streptomyces platensis transglutaminase (MtgA) produced by the Streptomyces lividans transformant 25-2 was purified by ammonium sulfate fractionation, followed by CM-Sepharose CL-6B fast flow, and blue-Sepharose fast flow chromatography. The purification factor was ~33.2-fold, and the yield was 65%. The molecular weight of the purified recombinant MtgA was 40.0 KDa as estimated by SDS-PAGE. The optimal pH and the temperature for the enzyme activity were 6.0 and 55 °C, respectively, and the enzyme was stable at pH 5.0–6.0 and at temperature 45–55 °C. Enzyme activity was not affected by Ca2+, Li+, Mn2+, Na+, Fe3+, K+, Mg2+, Al3+, Ba2+, Co2+, EDTA, or IAA but was inhibited by Fe2+, Pb2+, Zn2+, Cu2+, Hg2+, PCMB, NEM, and PMSF. Optimization of the fermentation medium resulted in a twofold increase of recombinant MtgA activity in both flasks (5.78 U/ml) and 5-l fermenters (5.39 U/ml). Large-scale productions of the recombinant MtgA in a 30-l air-lift fermenter and a 250-l stirred-tank fermenter were fulfilled with maximal activities of 5.36 and 2.54 U/ml, respectively.

Keywords

Large-scale production Properties Purification Recombinant Streptomyces platensis Transglutaminase 

Abbreviations

TGase

Transglutaminase

MtgA

Streptomyces platensis transglutaminase

SDS-PAGE

SDS-polyacrylamide gel electrophoresis

EDTA

Ethylenediamine tetraacetic acid

IAA

Iodoacetic acid

PCMB

p-Chloromercuribenzoate

PMSF

Phenyl methyl sulfonyl fluoride

NEM

N-ethylmaleimide

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Copyright information

© Society for Industrial Microbiology 2008

Authors and Affiliations

  • Shie-Jea Lin
    • 1
  • Yi-Fang Hsieh
    • 1
  • Li-An Lai
    • 1
  • Mei-Li Chao
    • 1
  • Wen-Shen Chu
    • 1
  1. 1.Bioresource Collection and Research CenterFood Industry Research and Development InstituteHsinchuTaiwan, ROC

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