Trypsin from the processing waste of the lane snapper (Lutjanus synagris) and its compatibility with oxidants, surfactants and commercial detergents.
A trypsin from the viscera of the lane snapper (Lutjanus synagris) was purified by heat treatment, fractionation with ammonium sulfate and affinity chromatography. The molecular weight of the enzyme was estimated to be 28.4 kDa (SDS-PAGE). The purified enzyme was capable of hydrolyzing the specific substrate for trypsin benzoyl-arginine-p-nitroanilide (BApNA) and was inhibited by benzamidine and tosyl lysine chloromethyl ketone (TLCK), synthetic trypsin inhibitors and phenylmethylsulfonyl fluoride (PMSF), which is a serine-protease inhibitor. The enzyme exhibited maximal activity at pH 9.0 and 45 degrees C and retained 100% of the activity after incubation at the optimal temperature for 30 min. At a concentration of 10 mM, activity was slightly activated by Ca(2+) and inhibited by the following ions in decreasing order: Cd(2+) > Hg(2+) > Cu(2+) > Zn(2+) > Al(3+). The effects of Ba(2+), K(1+) and Li(1+) proved to be less intensive. Using 1% (w/v) azocasein as substrate, the enzyme revealed high resistance (60% residual activity) when incubated with 10% H(2)O(2) for 75 min. The enzyme retained more than 80% activity after 60 min in the presence of different surfactants (Tween 20, Tween 80 and sodium choleate). The alkaline protease demonstrated compatibility with commercial detergents (7 mg/mL), such as Bem-te-vi, Surf and Ala, retaining more than 50% of initial activity after 60 min at 25 degrees C and 30 min at 40 degrees C. The thermostability and compatibility of this enzyme with commercial detergents suggest a good potentiality for application in the detergent industry.
Espósito
Talita S
TS
Laboratório de Enzimologia, Departamento de Bioquímica and Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco, Brazil.
Marcuschi
Marina
M
Amaral
Ian P G
IP
Carvalho
Luiz B
LB
Bezerra
Ranilson S
RS
D000818:Animals; D002412:Cations / Q000494:pharmacology; D002846:Chromatography, Affinity; D003902:Detergents / Q000737:chemistry / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability; D005590:Fractional Precipitation; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007220:Industrial Waste / Q000032:analysis; D007422:Intestines / Q000201:enzymology; D016877:Oxidants / Q000494:pharmacology*; D010473:Perciformes; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000494:pharmacology*; D014357:Trypsin / Q000302:isolation & purification* / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000494:pharmacology
D002412:Cations; D003902:Detergents; D007220:Industrial Waste; D016877:Oxidants; D013501:Surface-Active Agents; D014361:Trypsin Inhibitors; D006861:Hydrogen Peroxide; D014357:Trypsin