Purification and characterisation of a protease (tamarillin) from tamarillo fruit. (1st August 2018)

• Record Type: Journal Article
• Authors: Li, Zhao; Scott, Ken; Hemar, Yacine; Zhang, Huoming; Otter, Don
• Is Part Of: Food chemistry. Volume 256(2018)
• Page Start: 228
• Page End: 234
• Publication Date: 2018-08-01
• Subjects: Cyphomandra betacea -- Solanaceae -- Purification -- Characterisation -- De novo sequencing -- Subtilisin-like serine protease -- Tamarillin; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2018.02.091
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: A novel serine protease (tamarillin) purified from tamarillo fruit. High pH optimum at 11 and wide range of thermostability. Stable at most of environment condition but Hg 2+ in present. A subtilisin-like serine protease was identified using de novo sequencing. Abstract: A protease from tamarillo fruit ( Cyphomandra betacea Cav.) was purified by ammonium sulphate precipitation and diethylaminoethyl-Sepharose chromatography. Protease activity was determined on selected peak fractions using a casein substrate. Sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis showed that the peak with the highest protease activity consisted of one protein of molecular mass ca. 70 kDa. The protease showed optimal activity at pH 11 and 60 °C. It was sensitive to phenylmethylsulphonyl fluoride while ethylenediaminetetraacetic acid and p -chloromercuribenzoic acid had little effect on its activity, indicating that this enzyme was a serine protease. Hg 2+ strongly inhibited enzyme activity, possibly due to formation of mercaptide bonds with the thiol groups of the protease, suggesting that some cysteine residues may be located close to the active site. De novo sequencing strongly indicated that the protease was a subtilisin-like alkaline serine protease. The protease from tamarillo has been named 'tamarillin'.