Effect of laundry detergent formulation on the performance of alkaline phytoproteases

Background: Proteases constitute the largest product segment in the global industrial enzymes market; they are used in food, pharmaceutical, leather, textile, wood and detergent industries. Alkaline proteases improve the cleaning efficiency of detergents and represent one of the most successful applications of modern industrial biotechnology. The aim of this work was to study the performance of two alkaline phytoproteases, araujiain (Araujia hortorum Fourn.) and asclepain (Asclepias curassavica L.), for their potential application as additive in laundry detergent formulations. Results: The effect of pure non-ionic and ionic surfactants on proteolytic activity of araujiain and asclepain was analyzed measuring the remaining activity after 1 hr of incubation of those enzymes in aqueous solutions of surfactants at different concentrations (0.1, 0.4 and 1% v/v) and temperatures (25, 40 and 60ºC). Besides, the compatibility of the enzymes with six commercial laundry detergents was also studied measuring the remaining proteolytic activity at 37ºC after 1 hr. Commercial detergent components influenced in different ways on araujiain and asclepain, in spite of the similar behaviour of the two enzymes in buffer. In commercial detergent solutions, araujiain expressed between 60% and 140% of its remaining proteolytic activity in buffer (pH 8.5) at 37ºC after 1 hr, while asclepain, was practically inactivate in most of them at the same conditions. Conclusions: Proteolytic extract of Araujia hortorum fulfilled all the requirements for its application as additive for laundry detergents: high stability in a broad temperature range (25-70ºC), high activity in alkaline pH (7.5-9.5) and very good compatibility with the commercial detergent additives. Nevertheless, in spite of its high stability and activity in buffer, the proteolytic extract of Asclepias curassavica did not show the same performance than araujiain.Fil: Barberis, Sonia Esther. Universidad Nacional de San Luis. Facultad de Quimica, Bioquimica y Farmacia. Departamento de Farmacia. Laboratorio de Bromatologia; ArgentinaFil: Quiroga, Evelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET- San Luis. Instituto de Física Aplicada; ArgentinaFil: Barcia, Cristina. Universidad Nacional de San Luis; ArgentinaFil: Liggiere, Constanza. Universidad Nacional de la Plata. Facultad de Ciencias Exactas; Argentin

Effect of laundry detergent formulation on the performance of alkaline phytoproteases

Background: Proteases constitute the largest product segment in the global industrial enzymes market; they are used in food, pharmaceutical, leather, textile, wood and detergent industries. Alkaline proteases improve the cleaning efficiency of detergents and represent one of the most successful applications of modern industrial biotechnology. The aim of this work was to study the performance of two alkaline phytoproteases, araujiain (Araujia hortorum Fourn.) and asclepain (Asclepias curassavica L.), for their potential application as additive in laundry detergent formulations. Results: The effect of pure non-ionic and ionic surfactants on proteolytic activity of araujiain and asclepain was analyzed measuring the remaining activity after 1 hr of incubation of those enzymes in aqueous solutions of surfactants at different concentrations (0.1, 0.4 and 1% v/v) and temperatures (25, 40 and 60oC). Besides, the compatibility of the enzymes with six commercial laundry detergents was also studied measuring the remaining proteolytic activity at 37oC after 1 hr. Commercial detergent components influenced in different ways on araujiain and asclepain, in spite of the similar behaviour of the two enzymes in buffer. In commercial detergent solutions, araujiain expressed between 60% and 140% of its remaining proteolytic activity in buffer (pH 8.5) at 37oC after 1 hr, while asclepain, was practically inactivate in most of them at the same conditions. Conclusions: Proteolytic extract of Araujia hortorum fulfilled all the requirements for its application as additive for laundry detergents: high stability in a broad temperature range (25-70oC), high activity in alkaline pH (7.5-9.5) and very good compatibility with the commercial detergent additives. Nevertheless, in spite of its high stability and activity in buffer, the proteolytic extract of Asclepias curassavica did not show the same performance than araujiain.Centro de Investigación de Proteínas Vegetale

Asclepain cI, a proteolytic enzyme from Asclepias curassavica L., a south American plant, against Helicobacter pylori

Helicobacter pylori is a Gram negative bacterium most frequently associated with human gastrointestinal infections worldwide. The increasing occurrence of antibiotic-resistant isolates of H. pylori constitutes a challenge. The eradication of the microorganism is currently being considered a “high priority” by the World Health Organization (WHO). In this context, bioactive compounds found in natural products seem to be an effective therapeutic option to develop new antibiotics against the pathogen. In this study, we investigated the effect of asclepain cI, the main purified proteolytic enzyme of the latex of petioles and stems from Asclepia curassavica L. (Asclepiadaceae), a South American native plant, against H. pylori; in order to obtain a natural therapeutic adjuvant and a safe nutraceutical product. Asclepain cI showed antibacterial activity against reference strains and drug-resistant clinical isolates of H. pylori in vitro. A range of minimal inhibitory concentration (MIC) from 1 to 2 μg/ml and minimal bactericidal concentration (MBC) from 2 to 4 μg/ml was obtained, respectively. The action of asclepain cI on the transcription of omp18, ureA, flaA genes showed a significantly decreased expression of the selected pathogenic factors. Furthermore, asclepain cI did not induce toxic effects at the concentrations assayed. Asclepain cI could be considered a highly feasible option to be used as a natural therapeutic adjuvant and a safe nutraceutical product against H. pylori.Fil: Salinas Ibañez, Angel Gabriel. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; ArgentinaFil: Origone, Anabella Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Física Aplicada "Dr. Jorge Andrés Zgrablich". Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto de Física Aplicada "Dr. Jorge Andrés Zgrablich"; ArgentinaFil: Liggieri, Constanza Silvina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; ArgentinaFil: Barberis, Sonia Esther. Universidad Nacional de La Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Vega, Alba Edith. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; Argentin

Effect of laundry detergent formulation on the performance of alkaline phytoproteases

Background: Proteases constitute the largest product segment in the global industrial enzymes market; they are used in food, pharmaceutical, leather, textile, wood and detergent industries. Alkaline proteases improve the cleaning efficiency of detergents and represent one of the most successful applications of modern industrial biotechnology. The aim of this work was to study the performance of two alkaline phytoproteases, araujiain ( Araujia hortorum Fourn.) and asclepain ( Asclepias curassavica L.), for their potential application as additive in laundry detergent formulations. Results: The effect of pure non-ionic and ionic surfactants on proteolytic activity of araujiain and asclepain was analyzed measuring the remaining activity after 1 hr of incubation of those enzymes in aqueous solutions of surfactants at different concentrations (0.1, 0.4 and 1% v/v) and temperatures (25, 40 and 60\ubaC). Besides, the compatibility of the enzymes with six commercial laundry detergents was also studied measuring the remaining proteolytic activity at 37\ubaC after 1 hr. Commercial detergent components influenced in different ways on araujiain and asclepain, in spite of the similar behaviour of the two enzymes in buffer. In commercial detergent solutions, araujiain expressed between 60% and 140% of its remaining proteolytic activity in buffer (pH 8.5) at 37\ubaC after 1 hr, while asclepain, was practically inactivate in most of them at the same conditions. Conclusions: Proteolytic extract of Araujia hortorum fulfilled all the requirements for its application as additive for laundry detergents: high stability in a broad temperature range (25-70\ubaC), high activity in alkaline pH (7.5-9.5) and very good compatibility with the commercial detergent additives. Nevertheless, in spite of its high stability and activity in buffer, the proteolytic extract of Asclepias curassavica did not show the same performance than araujiain

Peptide synthesis: chemical or enzymatic

Peptides are molecules of paramount importance in the fields of health care and nutrition. Several technologies for their production are now available, among which chemical and enzymatic synthesis are especially relevant. The present review pretends to establish a non-biased appreciation of the advantages, potentials, drawbacks and limitations of both technologies. Chemical synthesis is thoroughly reviewed and their potentials and limitations assessed, focusing on the different strategies and challenges for large-scale synthesis. Then, the enzymatic synthesis of peptides with proteolytic enzymes is reviewed considering medium, biocatalyst and substrate engineering, and recent advances and challenges in the field are analyzed. Even though chemical synthesis is the most mature technology for peptide synthesis, lack of specificity and environmental burden are severe drawbacks that can in principle be successfully overcame by enzyme biocatalysis. However, productivity of enzymatic synthesis is lower, costs of biocatalysts are usually high and no protocols exist for its validation and scale-up, representing challenges that are being actively confronted by intense research and development in this area. The combination of chemical and enzymatic synthesis is probably the way to go, since the good properties of each technology can be synergistically used in the context of one process objective

Study of phytoproteases stability in aqueous-organic biphasic systems using linear free energy relationships

In this paper we study the effect of different water-immiscible organic solvents (benzene, toluene, 1-butanol, 1-octanol, dichloroethane, dichloromethane, diethyl ether, hexane, chlorobenzene, acetophenone, n-dodecane, trichloroethylene, ethyl acetate) on the stability (residual caseinolytic activity after 4 h) of soluble phytoproteases, such as araujiain, funastrain and papain in aqueous-organic biphasic systems. Besides, the effect of organic solvents on enzymatic catalysis was quantitatively studied by means of linear free energy relationships (LFERs). The organic solvents were characterized by several physicochemical properties, and multiple linear regression analysis (MLRA) together with non-linear regression were the methods used to search the relationships between the residual caseinolytic activity data and several physicochemical parameters. Those enzymes show much greater activity and stability in some biphasic media than in water. On the other hand, all developed correlations represented highly significant LFERs models and showed that non-specific polar and hydrophobic factors are of prime and approximately equal importance for the biocatalytic activity of araujiain, funastrain and papain in the studied biphasic systems, while the specific polar interactions are of little importance for activity. The results suggested that araujiain, funastrain and papain do not suffer unfolding in the studied biphasic media and they are able to retain their native or native-like configurations, though with altered characteristics or properties. This fact was demonstrated by means of a comparative FTIR spectroscopy study in both, buffer and biphasic media, for each studied enzyme.Facultad de Ciencias Exacta

Study of phytoproteases stability in aqueous-organic biphasic systems using linear free energy relationships

In this paper we study the effect of different water-immiscible organic solvents (benzene, toluene, 1-butanol, 1-octanol, dichloroethane, dichloromethane, diethyl ether, hexane, chlorobenzene, acetophenone, n-dodecane, trichloroethylene, ethyl acetate) on the stability (residual caseinolytic activity after 4 h) of soluble phytoproteases, such as araujiain, funastrain and papain in aqueous-organic biphasic systems. Besides, the effect of organic solvents on enzymatic catalysis was quantitatively studied by means of linear free energy relationships (LFERs). The organic solvents were characterized by several physicochemical properties, and multiple linear regression analysis (MLRA) together with non-linear regression were the methods used to search the relationships between the residual caseinolytic activity data and several physicochemical parameters. Those enzymes show much greater activity and stability in some biphasic media than in water. On the other hand, all developed correlations represented highly significant LFERs models and showed that non-specific polar and hydrophobic factors are of prime and approximately equal importance for the biocatalytic activity of araujiain, funastrain and papain in the studied biphasic systems, while the specific polar interactions are of little importance for activity. The results suggested that araujiain, funastrain and papain do not suffer unfolding in the studied biphasic media and they are able to retain their native or native-like configurations, though with altered characteristics or properties. This fact was demonstrated by means of a comparative FTIR spectroscopy study in both, buffer and biphasic media, for each studied enzyme.Facultad de Ciencias Exacta

Effect of laundry detergent formulation on the performance of alkaline phytoproteases

Background: Proteases constitute the largest product segment in the global industrial enzymes market; they are used in food, pharmaceutical, leather, textile, wood and detergent industries. Alkaline proteases improve the cleaning efficiency of detergents and represent one of the most successful applications of modern industrial biotechnology. The aim of this work was to study the performance of two alkaline phytoproteases, araujiain (Araujia hortorum Fourn.) and asclepain (Asclepias curassavica L.), for their potential application as additive in laundry detergent formulations. Results: The effect of pure non-ionic and ionic surfactants on proteolytic activity of araujiain and asclepain was analyzed measuring the remaining activity after 1 hr of incubation of those enzymes in aqueous solutions of surfactants at different concentrations (0.1, 0.4 and 1% v/v) and temperatures (25, 40 and 60oC). Besides, the compatibility of the enzymes with six commercial laundry detergents was also studied measuring the remaining proteolytic activity at 37oC after 1 hr. Commercial detergent components influenced in different ways on araujiain and asclepain, in spite of the similar behaviour of the two enzymes in buffer. In commercial detergent solutions, araujiain expressed between 60% and 140% of its remaining proteolytic activity in buffer (pH 8.5) at 37oC after 1 hr, while asclepain, was practically inactivate in most of them at the same conditions. Conclusions: Proteolytic extract of Araujia hortorum fulfilled all the requirements for its application as additive for laundry detergents: high stability in a broad temperature range (25-70oC), high activity in alkaline pH (7.5-9.5) and very good compatibility with the commercial detergent additives. Nevertheless, in spite of its high stability and activity in buffer, the proteolytic extract of Asclepias curassavica did not show the same performance than araujiain.Centro de Investigación de Proteínas Vegetale

Behavior of Araujiain , a new cysteine phytoprotease, in organic media with low water content

In this paper we studied the effect of different organic solvents (1-octanol, trichloroethylene, ethanol, ethyl acetate, tetrahydrofuran, cyclohexane, propanone, acetonitrile, dichloromethane, chlorobenzene, N,N-dimethylformamide, acetophenone, diethyl ether, methanol, ethylene glycol and toluene) with low and constant water content on substrate preferences, thermostability and stability (caseinolytic activity retention after 4 h) of proteases of Araujia hortorum Fourn. (Asclepiadaceae). The stability of araujiain was high in N,N-dimethylformamide and ethanol at 40\ub0C, but decreased at higher temperature. Araujiain substrates preferences in buffer Tris-HCl (pH 8), ethylene glycol and N,N-dimethylformamide exhibited different patterns, but the enzyme showed a high preference by glutamine derivative in all cases. According to FTIR spectroscopy studies, araujiain changed its secondary structure and as a consequence, it also changed its substrate preferences. This enzyme showed lower \u3b1-helical character and greater \u3b2-sheet folding in buffer than in organic media. A larger amount of antiparallel \u3b2-sheet residues indicates the formation of tighter intermolecular hydrogen bonds and enzymatic aggregates. These facts could explain the higher esterolytic activities, the greater stability and good hydrolytic potential of araujiain in some organic media such as N,N-dimethylformamide