Zika fever is a prevalent global disease caused by the Zika virus. The Aedes aegypti mosquito transmits the virus during the blood feeding process, making them ideal biological vectors. Blood meal digestion in the Ae. aegypti mosquito is required to complete the gonotrophic cycle. With the spread of the Zika virus and other arbosviruses during blood feeding, and with the recent appearance of Ae. aegypti mosquitoes in Northern California, it is crucial to slow the spread of the mosquito and the blood-borne diseases it transmits. The Ae. aegypti mosquito relies on digestive enzymes, known as serine proteases, to degrade and digest blood meal proteins into amino acids and nutrients required for the egg laying process. These proteases are expressed at different time points after a blood meal. The enzyme that will be discussed is the late phase protease AaSPVII. By understanding the biochemical process involved in blood meal metabolism and understanding the important roles midgut proteases play in this process, we could potentially target the proteases with small molecule inhibitors and disrupt fecundity. AaSPVII, one of the four abundantly expressed midgut serine proteases was solubly expressed using NEB SHuffle T7 competent E. coli (New England Biolabs). However, autocatalysis of the protease during the expression process has been observed due to the disappearance of the His6-tagged and BApNA activity. Currently, we are optimizing purification conditions in order to purify active mature AaSPVII protease. In addition, kinetic parameters and cleavage assays with hemoglobin and bovine serum albumin (BSA) will be compared to published results.

In vitro biochemical studies of the midgut serine protease AaSPVII of the Zika vector Aedes aegypti

253rd National Meeting (2017)

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