Phenoloxidase activity in the reproductive system and egg masses of the pulmonate gastropod, Biomphalaria glabrata.
Phenoloxidase (PO) activity in the albumen gland (AG) and egg masses (EM) of Biomphalaria glabrata was assessed using high-performance liquid chromatography combined with electrochemical detection and colorimetric techniques. Both AG and EM extracts catalyzed the hydroxylation of L-tyrosine (monophenol oxidase activity, MPO) and oxidation of L-dopa (diphenol oxidase activity, DPO). However, no PO activity was found in the ovotestis. Both MPO and DPO activities in AG and EM were significantly inhibited by 1-phenyl-2-thiourea and inactivated by boiling. Approximately 35% of MPO and 44% of DPO activities were detected in the soluble fraction of homogenized EM, in contrast to that of homogenized AG, which contained about 5% and 12%, respectively, of MPO and DPO activities. N-acetyl-dopamine, a diphenolic compound, enhanced the hydroxylation of tyrosine by the PO. The presence of both MPO and DPO activities also was confirmed by the accelerated accumulation of dopachrome during incubation of EM extracts with L-tyrosine in the absence of ascorbate. Temperature and pH optima for this enzyme were 30 degrees C and 7.5, respectively. The potential roles of PO in egg formation in B glabrata are discussed.
Bai
G
G
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison 53706, USA.
Li
J
J
Christensen
B M
BM
Yoshino
T P
TP
D000818:Animals; D001700:Biomphalaria / Q000201:enzymology*; D004156:Catechol Oxidase; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000031:analogs & derivatives / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007980:Levodopa / Q000378:metabolism; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D010063:Ovum / Q000201:enzymology*; D012098:Reproduction; D013696:Temperature; D014443:Tyrosine / Q000378:metabolism
C006374:N-acetyldopamine; D014443:Tyrosine; D007980:Levodopa; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase; D004298:Dopamine