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Characterization and large-scale production of recombinant Streptoverticillium platensis transglutaminase

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Journal of Industrial Microbiology & Biotechnology

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.

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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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Acknowledgments

This work was funded by the Ministry of Economic Affairs of the Republic of China (Grant 92-EC-17-A-17-R7-0563).

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Authors and Affiliations

  1. Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, 30062, Hsinchu, Taiwan, ROC

    Shie-Jea Lin, Yi-Fang Hsieh, Li-An Lai, Mei-Li Chao & Wen-Shen Chu

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  1. Shie-Jea Lin
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  2. Yi-Fang Hsieh
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  3. Li-An Lai
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Correspondence to Wen-Shen Chu.

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Shie-Jea Lin and Yi-Fang Hsieh contributed equally to this work.

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Lin, SJ., Hsieh, YF., Lai, LA. et al. Characterization and large-scale production of recombinant Streptoverticillium platensis transglutaminase. J Ind Microbiol Biotechnol 35, 981–990 (2008). https://doi.org/10.1007/s10295-008-0373-2

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  • DOI: https://doi.org/10.1007/s10295-008-0373-2

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