Application of supercritical carbon dioxide for preparation of starfish phospholipase A(2)

作者：Chun Byung Soo; Kishimura Hideki^*; Kanzawa Hiroto; Klomklao Sappasith; Nalinanon Sitthipong; Benjakul Soottawat; Ando Seiichi

来源：Process Biochemistry, 2010, 45(5): 689-693.

DOI：10.1016/j.procbio.2010.01.001

• Phospholipase A(2)
• Starfish
• Asterina pectinifera
• Defatting process
• Supercritical carbon dioxide
• Characteristics
• Food industry

Pyloric ceca of starfish (Asterina pectinifera) were treated by supercritical carbon dioxide (SCO2) to remove the lipids. Then, phospholipase A(2) (SC-PLA(2)) was extracted from the defatted powder and purified by a series of chromatographies including Sephacryl S-200, DEAE-cellulose, and Sephadex G-50. The purified SC-PLA(2) was nearly homogeneous in SDS-PAGE and native-PAGE. The molecular weight of the SC-PLA(2) was estimated as approximately 20,000. N-terminal amino acid sequence of the SC-PLA(2) was SVYQF. Temperature and pH optimums of the SC-PLA(2) were at around 50 degrees C and pH 9.0, respectively, and the enzyme activity was enhanced by sodium deoxycholate and 1 mM or higher concentration of Ca2+. The SC-PLA(2) was stimulated most by adding Ca2+ followed by Mg2+ and Co2+, while it was strongly inhibited by adding Zn2+ and EDTA. The SC-PLA(2) hydrolyzed phosphatidylcholine more effectively than phosphatidylethanolamine. These characteristics of the SC-PLA(2) were the same as those of the starfish PLA(2) (CM-PLA(2)) purified from the pyloric ceca defatted by chloroform-methanol (2:1, v/v) solution. Therefore, we concluded the SCO2 defatting process is useful as a substitute for organic solvent defatting process.

• 出版日期2010-5