Characterization of acetylcholinesterase from the gut of sea cucumber Stichopus japonicus

Hai Tao Wu, Dong Mei Li, Bei Wei Zhu, Ying Du, Xiao Qian Chai, Yoshiyuki Murata

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

An acetylcholinesterase was purified from the gut of sea cucumber Stichopus japonicus by anion exchange chromatography followed by gel filtration chromatography. The enzyme was purified 35. 49-fold with a total yield of 7. 73 %. The molecular mass of purified acetylcholinesterase was 68 kDa as revealed on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme displayed maximum activity at pH 7. 5 and 35 °C with acetylthiocholine iodide as substrate. The enzyme activity appeared to be stable over pH 6. 0-8. 0 and up to 40 °C. It displayed an apparent Michaelis-Menten behavior in the concentration range from 0. 1 to 0. 8 mM with Km values of 0. 62 mM for acetylthiocholine iodide and 2. 53 mM for butyrylthiocholine iodide. More than 95 % of acetylcholinesterase activity was inhibited by 1 mM eserine or 1,5-bis(4-allyldimethylammonium phenyl)-pentan-3-one dibromide (BW284C51), but only 19. 1 % of the activity was inhibited by tetraisopropylpyrophosphoramide (iso-OMPA) at the same concentration. On the basis of the substrate and inhibitor specificities, the purified enzyme appeared to be a true acetylcholinesterase. Nevertheless, the purified acetylcholinesterase exhibited insensitivity to substrate inhibition phenomenon. Its biochemical properties were compared with those reported for different species.

Original languageEnglish
Pages (from-to)303-311
Number of pages9
JournalFisheries Science
Volume79
Issue number2
DOIs
Publication statusPublished - 2013

Fingerprint

Apostichopus japonicus
Holothuroidea
iodide
acetylcholinesterase
digestive system
enzyme
iodides
substrate
chromatography
gel
enzymes
enzyme activity
inhibitor
electrokinesis
ion exchange
sodium
sulfate
fold
polyacrylamide gel electrophoresis
sea

Keywords

  • Acetylcholinesterase
  • Characterization
  • Gut
  • Purification
  • Sea cucumber Stichopus japonicus

ASJC Scopus subject areas

  • Aquatic Science

Cite this

Characterization of acetylcholinesterase from the gut of sea cucumber Stichopus japonicus. / Wu, Hai Tao; Li, Dong Mei; Zhu, Bei Wei; Du, Ying; Chai, Xiao Qian; Murata, Yoshiyuki.

In: Fisheries Science, Vol. 79, No. 2, 2013, p. 303-311.

Research output: Contribution to journalArticle

Wu, Hai Tao ; Li, Dong Mei ; Zhu, Bei Wei ; Du, Ying ; Chai, Xiao Qian ; Murata, Yoshiyuki. / Characterization of acetylcholinesterase from the gut of sea cucumber Stichopus japonicus. In: Fisheries Science. 2013 ; Vol. 79, No. 2. pp. 303-311.
@article{3f366cfa7419466da159de8f0198c507,
title = "Characterization of acetylcholinesterase from the gut of sea cucumber Stichopus japonicus",
abstract = "An acetylcholinesterase was purified from the gut of sea cucumber Stichopus japonicus by anion exchange chromatography followed by gel filtration chromatography. The enzyme was purified 35. 49-fold with a total yield of 7. 73 {\%}. The molecular mass of purified acetylcholinesterase was 68 kDa as revealed on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme displayed maximum activity at pH 7. 5 and 35 °C with acetylthiocholine iodide as substrate. The enzyme activity appeared to be stable over pH 6. 0-8. 0 and up to 40 °C. It displayed an apparent Michaelis-Menten behavior in the concentration range from 0. 1 to 0. 8 mM with Km values of 0. 62 mM for acetylthiocholine iodide and 2. 53 mM for butyrylthiocholine iodide. More than 95 {\%} of acetylcholinesterase activity was inhibited by 1 mM eserine or 1,5-bis(4-allyldimethylammonium phenyl)-pentan-3-one dibromide (BW284C51), but only 19. 1 {\%} of the activity was inhibited by tetraisopropylpyrophosphoramide (iso-OMPA) at the same concentration. On the basis of the substrate and inhibitor specificities, the purified enzyme appeared to be a true acetylcholinesterase. Nevertheless, the purified acetylcholinesterase exhibited insensitivity to substrate inhibition phenomenon. Its biochemical properties were compared with those reported for different species.",
keywords = "Acetylcholinesterase, Characterization, Gut, Purification, Sea cucumber Stichopus japonicus",
author = "Wu, {Hai Tao} and Li, {Dong Mei} and Zhu, {Bei Wei} and Ying Du and Chai, {Xiao Qian} and Yoshiyuki Murata",
year = "2013",
doi = "10.1007/s12562-012-0588-z",
language = "English",
volume = "79",
pages = "303--311",
journal = "Fisheries Science",
issn = "0919-9268",
publisher = "Springer Japan",
number = "2",

}

TY - JOUR

T1 - Characterization of acetylcholinesterase from the gut of sea cucumber Stichopus japonicus

AU - Wu, Hai Tao

AU - Li, Dong Mei

AU - Zhu, Bei Wei

AU - Du, Ying

AU - Chai, Xiao Qian

AU - Murata, Yoshiyuki

PY - 2013

Y1 - 2013

N2 - An acetylcholinesterase was purified from the gut of sea cucumber Stichopus japonicus by anion exchange chromatography followed by gel filtration chromatography. The enzyme was purified 35. 49-fold with a total yield of 7. 73 %. The molecular mass of purified acetylcholinesterase was 68 kDa as revealed on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme displayed maximum activity at pH 7. 5 and 35 °C with acetylthiocholine iodide as substrate. The enzyme activity appeared to be stable over pH 6. 0-8. 0 and up to 40 °C. It displayed an apparent Michaelis-Menten behavior in the concentration range from 0. 1 to 0. 8 mM with Km values of 0. 62 mM for acetylthiocholine iodide and 2. 53 mM for butyrylthiocholine iodide. More than 95 % of acetylcholinesterase activity was inhibited by 1 mM eserine or 1,5-bis(4-allyldimethylammonium phenyl)-pentan-3-one dibromide (BW284C51), but only 19. 1 % of the activity was inhibited by tetraisopropylpyrophosphoramide (iso-OMPA) at the same concentration. On the basis of the substrate and inhibitor specificities, the purified enzyme appeared to be a true acetylcholinesterase. Nevertheless, the purified acetylcholinesterase exhibited insensitivity to substrate inhibition phenomenon. Its biochemical properties were compared with those reported for different species.

AB - An acetylcholinesterase was purified from the gut of sea cucumber Stichopus japonicus by anion exchange chromatography followed by gel filtration chromatography. The enzyme was purified 35. 49-fold with a total yield of 7. 73 %. The molecular mass of purified acetylcholinesterase was 68 kDa as revealed on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme displayed maximum activity at pH 7. 5 and 35 °C with acetylthiocholine iodide as substrate. The enzyme activity appeared to be stable over pH 6. 0-8. 0 and up to 40 °C. It displayed an apparent Michaelis-Menten behavior in the concentration range from 0. 1 to 0. 8 mM with Km values of 0. 62 mM for acetylthiocholine iodide and 2. 53 mM for butyrylthiocholine iodide. More than 95 % of acetylcholinesterase activity was inhibited by 1 mM eserine or 1,5-bis(4-allyldimethylammonium phenyl)-pentan-3-one dibromide (BW284C51), but only 19. 1 % of the activity was inhibited by tetraisopropylpyrophosphoramide (iso-OMPA) at the same concentration. On the basis of the substrate and inhibitor specificities, the purified enzyme appeared to be a true acetylcholinesterase. Nevertheless, the purified acetylcholinesterase exhibited insensitivity to substrate inhibition phenomenon. Its biochemical properties were compared with those reported for different species.

KW - Acetylcholinesterase

KW - Characterization

KW - Gut

KW - Purification

KW - Sea cucumber Stichopus japonicus

UR - http://www.scopus.com/inward/record.url?scp=84874791388&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84874791388&partnerID=8YFLogxK

U2 - 10.1007/s12562-012-0588-z

DO - 10.1007/s12562-012-0588-z

M3 - Article

VL - 79

SP - 303

EP - 311

JO - Fisheries Science

JF - Fisheries Science

SN - 0919-9268

IS - 2

ER -