A simple method for Isolation of serine protease inhibitor from Siamese land snail Cryptozona (Cryptozona siamensis) mucus

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Phichaya Khamai
Pariyaphat Ngandee
Ketpaillin Chimong
Rungthiwa Srimora
Pannawich Siriwechviriya
Widsanusan Chartarrayawadee
Kritchai Poonchareon


Background: Siamese land snail is the single shell snail which has become widespread very quickly in Thailand and other parts of the world.  It is considered to be a pest by most farmers since it destroys parts of plants such as leaves, vegetables and fruit by consuming them and leaving behind the line of sticky mucus over the plants.  This mucus is believed to be the snail’s defense mechanism, such as lubricant, for wound healing as well as a response against the snail’s predators. That the land snail releases mucus in response to the predator has been hypothesized, however, screening for protease inhibitor has still not substantiated this.  On the basis of the prevailing assumptions, through biochemical, and biophysical isolation we investigated the proteinase inhibitor functions of the protein from the mucus of the Siamese land snail.

Objectives: This study aimed to find a potential protease inhibitor from the mucus of the local Northern Thailand land snail (Cryptozona siamensis) together with the biochemical characterization of its general properties.

Materials and methods: Land snail extract collected from Phayao Province, Thailand, was initially subjected to trypsin inhibitory activity assay using alzoalbumin as substrates. The detection of its inhibition activity was assessed by spectrophotometry. Additionally, the molecular size was observed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE).

Results: Land snail (Cryptozona siamensis) mucus contained serine protease inhibitory activity. The molecular size of the inhibitor was suspected to be about 10.5 kD.  Interestingly, its inhibitory property was retained after heat inactivation at 100 ᵒC for 20 min.

Conclusion: A novel heat-tolerant serine protease inhibitor from acetone extraction of land snail mucus was characterized as a small peptide with tolerance to organic solvent (acetone) and high temperature, which could be used as a new peptide protease inhibitor targeting serine protease that may benefit to scientific and medical fields.


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Khamai, P., Ngandee, P., Chimong, K., Srimora, R., Siriwechviriya, P., Chartarrayawadee, W., & Poonchareon, K. (2020). A simple method for Isolation of serine protease inhibitor from Siamese land snail Cryptozona (Cryptozona siamensis) mucus. Journal of Associated Medical Sciences, 53(3), 78-83. Retrieved from https://he01.tci-thaijo.org/index.php/bulletinAMS/article/view/243099
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