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Characterization of Neutral Lipase BT-1 Isolated from the Labial Gland of Bombus terrestris Males
We studied the temporal dynamics of lipolytic activity in crude extracts from the cephalic part of Bombus terrestris labial glands. Extracts from 3-day-old males displayed the highest lipolytic activity. The highest lipase gene expression level was observed in freshly emerged bumblebees, and both gene expression and lipase activity were lower in bumblebees older than 3 days. Lipase was purified from labial glands, further characterized and named as BT-1. The B. terrestris orthologue shares 88% sequence identity with B. impatiens lipase HA. The molecular weight of B. terrestris lipase BT-1 was approximately 30 kDa, the pH optimum was 8.3, and the temperature optimum was 50°C. Lipase BT-1 showed a notable preference for C8-C10 p-nitrophenyl esters, with the highest activity toward p-nitrophenyl caprylate (C8). The Michaelis constant (K[SUB]m[/SUB]) and maximum reaction rate (V[SUB]max[/SUB]) for p-nitrophenyl laurate hydrolysis were K[SUB]m[/SUB] = 0.0011 mM and V[SUB]max[/SUB] = 0.15 U/mg.
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080066
We studied the temporal dynamics of lipolytic activity in crude extracts from the cephalic part of Bombus terrestris labial glands. Extracts from 3-day-old males displayed the highest lipolytic activity. The highest lipase gene expression level was observed in freshly emerged bumblebees, and both gene expression and lipase activity were lower in bumblebees older than 3 days. Lipase was purified from labial glands, further characterized and named as BT-1. The B. terrestris orthologue shares 88% sequence identity with B. impatiens lipase HA. The molecular weight of B. terrestris lipase BT-1 was approximately 30 kDa, the pH optimum was 8.3, and the temperature optimum was 50°C. Lipase BT-1 showed a notable preference for C8-C10 p-nitrophenyl esters, with the highest activity toward p-nitrophenyl caprylate (C8). The Michaelis constant (K[SUB]m[/SUB]) and maximum reaction rate (V[SUB]max[/SUB]) for p-nitrophenyl laurate hydrolysis were K[SUB]m[/SUB] = 0.0011 mM and V[SUB]max[/SUB] = 0.15 U/mg.
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080066
