Chapter 13: Acacia caven (Mol.) Molina pollen proteases. Application to the peptide synthesis and to laundry detergents.

CRISTINA BARCIA; EVELINA QUIROGA; CARLOS ARDANAZ; GUSTAVO QUIROGA; SONIA BARBERIS

Biochemical Engineering. Series: Biotechnology in Agriculture, Industry and Medicine (Fabián E. Dumont and Jack A. Sacco, Editors).

Nova Science Publisher. http://www.novapublishers.com.

Lugar: Hauppauge, NY; Año: 2009; p. 293 - 316

It is known that the proteases have applications in several industrial processes such us leather processing, laundry detergents, producing of protein hydrolysates and food processing, as well as in the peptide synthesis in non conventional media. The application of proteases as catalyst of short oligopeptides in aqueous -organic media, have received a great deal  attentionas a viable alternative to chemical approach because of their remarkable characteristics. On the other hand, alkaline proteases have also been used to improve the cleaning efficiency of detergents. Detergent enzymes account for about 30% of the t otal worldwide enzyme production and represent one of the largest and most successful applications of modern industrial biotechnology. The aim of this work was to study the performance of proteolytic enzymes of Acacia caven (Mol.) Molina pollen for its potential application as an additive in various laundry detergents formulations and as catalyst of the peptide synthesis in aqueous-organic media. Pollen grains (35 mg/ml) were suspended in 0.1M Tris-HCl buffer pH 7.4 and slowly shaken for 2 h at 25ºC. Then, the slurry was centrifugated for 30 min at 8000 rpm and the supernatant (crude enzyme extract, CE) was tested in protein content (Bradford’s method) and proteolytic activity (using BAPNA and  Z-Ala-pNO as substrates). A partial characterization of  Acacia c aven CE was carried out: enzyme extract displayed maximum proteolytic activity at pH 8 and  35-40º C; it showed remarkable thermal stability after 1.5 h at 25-40º C but it decreased as long as temperature increased to 60º C. On the other hand, the enzyme extract was incubated with different surfactants and commercial laundry detergents at 25-60° C during 30 min and 1h; and it showed high stability and compatibility with them. The peptide synthesis catalyzed by  Acacia  caven CE was carried out in a mixture of  0.1M Tris-HCl buffer  pH 8.5 and ethyl acetate (50:50 ratio) at 37° C using  2-mercaptoethanol as activator and TEA as neutralizing agent of the amino component (Phe-OMe.HCl). Carboxylic components were selected in base of the highest preference of CE. The identification of synthesized peptide products was carried out by HPLC-MS.  According to the obtained results, this work contributes with a new variety of phytoprotease useful as catalyst of the peptide synthesis and as additive of laundry detergents.