Isolation and Biochemical Characteristics Analyses of Phenoloxidases (POs) in Three Cultured Mollusk Species

• 作者: SONG Xiangdi JIANG Jingwei XING Jing ZHAN Wenbin
• 来源期刊: 中国海洋大学学报（自然科学英文版）
• 年，卷（期）: 2022,（2）
• 所属期刊栏目: Life and Fisheries Sciences
• 页码范围: 465-472
• 页数: 8页
• 语种: 英文

摘要

Phenoloxidases (POs) are a group of copper proteins including tyrosinase, catecholase and laccase, which play crucial roles in the innate immune response of mollusks. In this research, POs were studied in cultured mollusk species, including scallop Chla- mys farreri, abalone Haliotis discus hannai and clam Scapharca subcrenata. The POs were isolated from hemocytes using linear- gradient native-PAGE combined with catechol staining. The PO activities and their characters were investigated. The molecular mass of PO in C. farreri was 576 kDa, and it was 228 kDa in H. discus hannai. In S. subcrenata, four POs were detected and their mole- cular masses were 391 kDa, 206 kDa, 174 kDa and < 67 kDa, which were named as 391-PO, 206-PO, 174-PO and s-PO, respectively. Ki- netic analyses indicated that all of the POs, except for 391-PO had higher affinity to L-DOPA and catechol than to hydroquinone and dopamine. However, all of the POs failed to oxidize tyrosine. The effects of divalent metal ions on POs' activities were assayed, in- cluding Fe2+, Mg2+, Zn2+, Mn2+, Cu2+ and Ca2+ from FeCl2, MgSO4, ZnSO4, MnCl2, CuSO4 and CaCl2. The POs were inhibited by Fe2+ at all determined concentrations. Additionally, the inhibition assay showed that all of the POs were inhibited by cysteine, ascor-bic acid, sodium sulfite, citric acid, ethylenediaminetetraacetic acid disodium (EDTA) and sodium diethyldithiocarbamate (DETC). The inhibition effects of critric acid and EDTA are dose-dependent. H. discus hannai PO and 391-PO were slightly inhibited by sodium azide, and H. discus hannai PO, 391-PO and 174-PO were slightly inhibited by thiourea. In conclusion, the POs in the three cultured mollusks are copper-containing laccase-type phenoloxidases with similar biochemical characteristics even though their molecular masses are different.