Plant proteases are capable of performing several functions in biological systems, and their use is attractive for biotechnological process due to their interesting catalytic properties. Bromelia pinguin (aguama) is a wild abundant natural resource in several regions of Central America and the Caribbean Islands but is underutilized. Their fruits are rich in proteases with properties that are still unknown, but they represent an attractive source of enzymes for biotechnological applications. Thus, the proteolytic activity in enzymatic crude extracts (CEs) from wild B. pinguin fruits was partially characterized. Enzymes in CEs showed high proteolytic activity at acid (pH 2.0-4.0) and neutral alkaline (pH 7.0-9.0) conditions, indicating that different types of active proteases are present. Proteolytic activity inhibition by the use of specific protease inhibitors indicated that aspartic, cysteine, and serine proteases are the main types of proteases present in CEs. Activity at pH 3.0 was stable in a broad range of temperatures (25-50 °C) and retained its activity in the presence of surfactants (SDS, Tween-80), reducing agents (DTT, 2-mercapoethanol), and organic solvents (methanol, ethanol, acetone, 2-propanol), which suggests that B. pinguin proteases are potential candidates for their application in brewing, detergent, and pharmaceutical industries.

Agave tequilana Weber azul variety is a valuable source of fructans. In this study, a response surface methodology was employed to optimize the extraction yield of fructans from agave pines. A Box Behnken factorial design was applied to evaluate the effects of several conditions such as extraction temperature, water raw material ratio and extraction time on extraction yield. Under the optimum extraction conditions (extraction temperature of 79.1 °C, water raw material ratio of 5.13 mL/g, and extraction time of 1.48 h) fructans extraction yield was 83.12%. The chromatographic profile of the agave crude extract presented four peaks, out of which fructans were the most preponderant (~87%). The obtained results suggest that the response surface methodology is adequate to optimize fructans extraction from Agave tequilana Weber azul variety.Keywords: fructans; agave; extraction yield; optimization; response surface methodology.Practical Application: Response surface methodology allows the optimization of fructans extraction from Agave azul.

High amounts of rich-protein liquid wastes are produced during seafood processing. The e uent called stickwater resulting from the processing of Paci c thread herring (Ophistonema libertate) into shmeal, was evaluated as protein source to produce bioactive protein hydrolysates by using Alcalase as enzyme source. MethodsThe effect of degree of hydrolysis on biochemical properties (proximate analysis, molecular weight, amino acid composition) and antioxidant and antihypertensive activities of stickwater protein hydrolysates obtained with Alcalase was determined. ResultsDegree of hydrolysis (DH) of samples (5, 10 15 and 20%) in uences its biochemical and bioactive properties. The maximum ABTS and FRAP activity values (P < 0.05) were exhibited by hydrolysates at 15% DH (EC 50 = 2.8mg/mL and TEAC = 1.16 ± 0.03 mM TE/mg, respectively). Whereas the highest DPPH scavenging activity (P < 0.05) was found for hydrolysates at 5 and 10% of DH (EC 50 = 34.7 and 37 mg/mL respectively). Furthermore, enzymatic hydrolysis enhanced angiotensin converting enzyme (ACE)inhibitory activity, being those at 5 and 10% of DH, which exhibited lower IC 50 values (P < 0.05) compared to non-hydrolyzed stickwater. Peptide distribution of protein hydrolysates at < 1.35 kDa was in a range of 47 to 62% of total peptides and the presence of amino acids related to antioxidant activity such as His, Lys, Met, Tau, Tyr and Trp was detected in stickwater and protein hydrolysates. ConclusionThe production of protein hydrolysates from Paci c thread herring stickwater, represents an alternative to obtain added-value products with potential antioxidant and antihypertensive activity. Statement Of NoveltyStickwaters are shery e uents with a high amount of dissolved protein containing a good pro le of essential amino acids. Taking in account that biochemical and bioactive properties are dependent of the extent of protein hydrolysis, in the present study the effect of degree of hydrolysis of herring stickwater was evaluated on antioxidant and antihypertensive activities, which, to the best of our knowledge, in other

En este estudio se realizó la caracterización parcial de los extractos enzimáticos de frutos de Bromelia pinguin basado en su actividad coagulante de la leche, actividad caseinolítica y su inhibición con el uso de inhibidores específicos. Los extractos enzimáticos crudos mostraron una máxima actividad coagulante de la leche entre 65-85ºC, correspondiente a 3.99 unidades Soxhlet·mL-1. Esto indica que 1 mL del extracto enzimático fue suficiente para coagular 400 mL de leche en los primeros 40 min. El efecto del pH, estudios de inhibición y análisis por zimografía indican que la actividad caseinolítica se debe a la presencia de diferentes proteasas tipo cisteína y serina. El análisis por SDS-PAGE evidenció que las proteasas presentes en el extracto hidrolizan preferentemente la fracción κ-caseína durante los primeros 30 minutos, obteniéndose un perfil hidrolítico similar al que se obtiene cuando se utiliza la enzima comercial quimosina; no obstante se observa que después de este tiempo de incubación otras caseínas también fueron susceptibles a la acción proteolítica. Bajo condiciones convencionales de coagulación, las proteasas tipo cisteína y serina presentes en el extracto de B. pinguin contribuyeron en mayor grado al proceso de coagulación. Bromelia pinguin representa una opción atractiva como fuente de proteasas para su aprovechamiento y aplicación en procesos biotecnológicos que requieren el uso de proteasas.

Purpose High amounts of rich-protein liquid wastes are produced during seafood processing. The effluent called stickwater resulting from the processing of Pacific thread herring (Ophistonema libertate) into fishmeal, was evaluated as protein source to produce bioactive protein hydrolysates by using Alcalase as enzyme source. Methods The effect of degree of hydrolysis on biochemical properties (proximate analysis, molecular weight, amino acid composition) and antioxidant and antihypertensive activities of stickwater protein hydrolysates obtained with Alcalase was determined. Results Degree of hydrolysis (DH) of samples (5, 10 15 and 20%) influences its biochemical and bioactive properties. The maximum ABTS and FRAP activity values (P < 0.05) were exhibited by hydrolysates at 15% DH (EC50 = 2.8mg/mL and TEAC = 1.16 ± 0.03 mM TE/mg, respectively). Whereas the highest DPPH scavenging activity (P < 0.05) was found for hydrolysates at 5 and 10% of DH (EC50 = 34.7 and 37 mg/mL respectively). Furthermore, enzymatic hydrolysis enhanced angiotensin converting enzyme (ACE)-inhibitory activity, being those at 5 and 10% of DH, which exhibited lower IC50 values (P < 0.05) compared to non-hydrolyzed stickwater. Peptide distribution of protein hydrolysates at < 1.35 kDa was in a range of 47 to 62% of total peptides and the presence of amino acids related to antioxidant activity such as His, Lys, Met, Tau, Tyr and Trp was detected in stickwater and protein hydrolysates. Conclusion The production of protein hydrolysates from Pacific thread herring stickwater, represents an alternative to obtain added-value products with potential antioxidant and antihypertensive activity.