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A trypsin‐like protease in rice green semi‐looper, Naranga aenescens moore (lepidoptera: Noctuidae): purification and characterization
Author(s) -
Zibaee Arash,
Bandani Ali Reza,
FazeliDinan Mahmoud,
Zibaee Idin,
Sendi Jalal Jalali,
Maleki Feizollah A.
Publication year - 2011
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.20435
Subject(s) - proteases , midgut , noctuidae , trypsin , protease , biology , lepidoptera genitalia , serine protease , biochemistry , enzyme , serine , chymotrypsin , botany , larva
The rice green semi looper, Naranga aenescens Moore (Lepidoptera: Noctuidae) causes severe damage to rice fields in Eastern Asia and Middle East. We demonstrate that two types of serine proteases are active in the midgut of the third instar larvae of N. aenescens , but trypsin‐like proteases are considerably more active than chymotrypsin‐like proteases. To develop better control strategies, purification and biochemical characterization of a major trypsin‐like digestive protease from the midgut of the third instar larvae of N. aenescens was achieved by gel filtration and anion exchange chromatography. After the final purification step, the enzyme was purified 9.62‐fold with a recovery of 16.1% and a specific activity of 4.12 U/mg protein and a molecular mass of approximately 88.5 kDa. Biochemical characterization indicated that the purified protease had highest activity at pH 10 and 30°C and was stable for up to 6 h under those conditions. Divalent cations, especially Ca 2+ , Mg 2+ , and Cu 2+ , increased the enzyme activity and synthetic inhibitors that target trypsin‐like activity caused a significant reduction in caseinolytic activity. These data may be used to develop inhibitors that decrease the damage of N. aenescens to rice cultivars in the field. © 2011 Wiley Periodicals, Inc.