title: Influence of ammonium salts on the lipase/esterase activity assay using p-nitrophenyl esters as substrates.
abstract: p-Nitrophenyl esters with a short-chain carboxylic group, such as p-nitrophenyl acetate (p-NPA) and p-nitrophenyl butyrate (p-NPB), could be effectively hydrolyzed by ammonium salts. p-Nitrophenyl esters were usually used as substrates to assay the lipase/esterase activity. Ammonium sulfate precipitation was often used to purify proteins, and some ammonium salts were usually used as nitrogen sources or inorganic salts for the lipase/esterase production. To study the effect of ammonium salts on the assay of the lipase/esterase activity, the contributing factors of hydrolysis of p-NPA/p-NPB catalyzed by ammonium salts were investigated. The lipase activities were compared in the presence and absence of ammonium sulfate. The hydrolysis reaction could be catalyzed under neutral and alkaline circumstances. The hydrolysis rate increased with the increase in the reaction temperature or the concentration of ammonium ion. When p-NPA was employed as the substrate for the analysis of the lipase/esterase activity, the effect of ammonium sulfate on the analysis could be neutralized by setting a control when the concentration of ammonium sulfate was less than 40% saturation. However, when the concentration of ammonium sulfate increased from 40% to 100% saturation, the enzyme activities decreased about 13-40%, which could not be ignored for accurate analysis of the enzyme activity.
authors.affiliation: College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, PR China.
authors.forename: Hong
authors.initials: H
authors.lastname: De Yan
authors.forename: Yin Jun
authors.initials: YJ
authors.lastname: Zhang
authors.forename: Hong Cai
authors.initials: HC
authors.lastname: Liu
authors.forename: Jian Yong
authors.initials: JY
authors.lastname: Zheng
authors.forename: Zhao
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry*; D001681:Biological Assay / Q000379:methods; D002087:Butyrates / Q000737:chemistry*; D004950:Esterases / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry*; D009596:Nitrophenols / Q000737:chemistry*; D012492:Salts / Q000737:chemistry*; D013696:Temperature
chemical_list: D002087:Butyrates; D009596:Nitrophenols; D012492:Salts; C033592:4-nitrophenyl butyrate; C008642:4-nitrophenyl acetate; D004950:Esterases; D008049:Lipase; D000645:Ammonium Sulfate