Study on the Properties of Recombinant Prolyl Endopeptidase from Abalone (<i>Haliotis discus hannai</i>) Muscle

Wanyu LI; Yue LI; Ling WENG; Shoufeng CHEN; Yulei CHEN; Guangming LIU; Minjie CAO

doi:10.13386/j.issn1002-0306.2020060196

Volume 42 Issue 7

Mar. 2021

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Science and Technology of Food Industry > 2021 > 42(7): 129-135. > DOI: 10.13386/j.issn1002-0306.2020060196

LI Wanyu, LI Yue, WENG Ling, et al. Study on the Properties of Recombinant Prolyl Endopeptidase from Abalone ( Haliotis discus hannai ) Muscle[J]. Science and Technology of Food Industry, 2021, 42(7): 129−135. (in Chinese with English abstract). doi: 10.13386/ j.issn1002-0306.2020060196.

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Study on the Properties of Recombinant Prolyl Endopeptidase from Abalone (Haliotis discus hannai) Muscle

Wanyu LI^1,,
Yue LI¹,
Ling WENG^{1, 2},
Shoufeng CHEN¹,
Yulei CHEN^{1, 2},
Guangming LIU^{1, 2},
Minjie CAO^{1, 2, ,}

1.
College of Food and Biological Engineering, Jimei University, Xiamen 361021, China
2.
Collaborative Innovation Center of Marine Food Deep Processing, Dalian Polytechnic University, Dalian 116034, China

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Received Date: June 15, 2020
Available Online: January 27, 2021

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Abstract

Abstract

In order to study the enzymatic characteristics and structure of prolyl endopeptidase from Haliotis discus hannai (Hdh-PEP), recombinant PEP was cloned and highly expressed in E.coli. Hdh-PEP with molecular weight of 85 kDa was successfully purified and its surface hydrophobicity was greatly affected by pH at low value (pH2～6) and temperature (20～60 ℃). Amino acid sequence homology analysis showed that there were three highly conserved sequences in the catalytic domain of Hdh-PEP: Seq 1: K-D-G-T-K/R-I-P, Seq 2: Y-G-Y-G-G-F and Seq 3: I-R-G-G-E-Y/F. Kinetic study revealed that the K_m and k_cat of Hdh-PEP were 5.32 μmol/L and 15.7 s⁻¹, respectively. Specific inhibitors SUAM-14746 and ZPP of PEP had strong inhibition on Hdh-PEP activity, and serine protease inhibitor (PMSF) also exhibited inhibition. A high specific polyclonal antibody toward Hdh-PEP was prepared which could be applied for the detection of native PEP in abalone muscle. The successful in vitro expression of Hdh-PEP and preparation of a specific polyclonal antibody against Hdh-PEP provided an important reference for subsequent investigation on Hdh-PEP.
- prolyl endopeptidase,
- Haliotis discus hannai,
- surface hydrophobicity,
- polyclonal antibody

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References (30)

References

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