Effect of Pectinolytic Enzyme Pretreatment on the Clarification of Cranberry Juice by Ultrafiltration

Cranberries, mainly processed as juice, have garnered interest over the past decade due to their high content of phytochemical compounds related to promising health benefits. To meet consumer expectations, a juice clarification step is usually incorporated to remove suspended solids. The use of pectinolytic enzyme and membrane processes are commonly applied to the production of clarified juices, but no studies have been done on cranberry juice. In this study, the effects of 60 (D60) and 120 min (D120) of depectinization by pectinolytic enzymes coupled to clarification by ultrafiltration (UF) (membrane molecular weight cut-off (MWCO) of 50, 100 and 500 kDa) was evaluated on the filtration performance, membrane fouling and cranberry juice composition. Compared to fresh juice, depectinization for 60 and 120 min reduced the UF duration by 16.7 and 20 min, respectively. The best filtration performance, in terms of permeate fluxes, was obtained with the 500 kDa MWCO UF membrane despite the highest total flux decline (41.5 to 57.6%). The fouling layer at the membrane surface was composed of polyphenols and anthocyanins. Compared to fresh juice, anthocyanin decreased (44% and 58% for D60 and D120, respectively) in depectinized juices whereas proanthocyanidin (PAC) content increased by 16%. In view of the industrial application, a 60 min depectinization coupled to clarification by a 500 kDa UF membrane could be viewed as a good compromise between the enhancement of filtration performance and the loss of polyphenols and their fouling at the membrane surface.

From secretion in Pichia pastoris to application in apple juice processing: Exo-polygalacturonase from Sporothrix schenckii 1099-18

Background: Polygalacturonases are a group of enzymes under pectinolytic enzymes related to enzymes that hydrolyse pectic substances. Polygalacturonases have been used in various industrial applications such as fruit juice clarification, retting of plant fibers, wastewater treatment drinks fermentation, and oil extraction. Objectives: The study was evaluated at the heterologous expression, purification, biochemical characterization, computational modeling, and performance in apple juice clarification of a new exo-polygalacturonase from Sporothrix schenckii 1099-18 (SsExo-PG) in Pichia pastoris. Methods: Recombinant DNA technology was used in this study. Two different pPIC9K plasmids were constructed with native signal sequence-ssexo-pg and alpha signal sequence-ssexo-pg separately. Protein expression and purification performed after plasmids transformed into the Pichia pastoris. Biochemical and structural analyses were performed by using pure SsExo-PG. Results: The purification of SsExo-PG was achieved using a Ni-NTA chromatography system. The enzyme was found to have a molecular mass of approximately 52 kDa. SsExo-PG presented as stable at a wide range of temperature and pH values, and to be more storage stable than other commercial pectinolytic enzyme mixtures. Structural analysis revealed that the catalytic residues of SsExo-PG are somewhat similar to other Exo-PGs. The KM and kcat values for the degradation of polygalacturonic acid (PGA) by the purified enzyme were found to be 0.5868 µM and 179 s-1, respectively. Cu2+ was found to enhance SsExo-PG activity while Ag2+ and Fe2+ almost completely inhibited enzyme activity. The enzyme reduced turbidity up to 80% thus enhanced the clarification of apple juice. SsExo-PG showed promising performance when compared with other commercial pectinolytic enzyme mixtures. Conclusion: The clarification potential of SsExo-PG was revealed by comparing it with commercial pectinolytic enzymes. The following parameters of the process of apple juice clarification processes showed that SsExo-PG is highly stable and has a novel performance.

A Review on Production of Polygalacturonase Using Various Organisms and Its Applications

Polygalacturonase is a pectinolytic enzyme that catalyses the hydrolytic cleavage of the polygalacturonic linkage chain. An enzyme is a polygalacturonase is expressed in fruits. The polygalacturonase produced from various organisms isolated from various fruits. The solid-state fermentation was used in the production of polygalacturonase. The production of PG was found at various incubation period and pH and temperature are using fruits as best nitrogen and carbon sources. Although they have other parts of the genome they are active in the fruit. Peak expression requires a full range of promoter and saturation lines. Based on the physicochemical properties of the purified enzymes, this enzyme possesses great potential for industrial and biotechnological application such as oil extraction, fruit clarifications.

Protease-, Pectinase- and Amylase- Producing Bacteria from a Kenyan Soda Lake

Background:Alkaline enzymes are stable biocatalysts with potential applications in industrial technologies that offer high quality products.Objective:The growing demand for alkaline enzymes in industry has enhanced the search for microorganisms that produce these enzymes.Methods:Eighteen bacterial isolates from Lake Bogoria, Kenya, were screened for alkaline proteases, pectinases and amylases; characterized and subjected to quantitative analysis of the enzymes they produced.Results:The screening analysis ranked 14, 16 and 18 of the bacterial isolates as potent producers of alkaline proteases, pectinases and amylases, respectively. The isolates were classified into two groups: Group 1 (16 isolates) were facultatively alkaliphilicB. haloduranswhile group 2 (2 isolates) were obligately alkaliphilicB. pseudofirmus. Further analysis revealed that group 1 isolates were divided into two sub-groups, with sub-group I (4 isolates) being a phenotypic variant sub-population of sub-group II (12 isolates). Variation between the two populations was also observed in their enzymatic production profilese.g. sub-group I isolates did not produce alkaline proteolytic enzymes while those in sub-group II did so (0.01-0.36 U/ml). Furthermore, they produced higher levels of the alkaline pectinolytic enzyme polygalacturonase (0.12-0.46 U/ml) compared to sub-group II isolates (0.05-0.10 U/ml), which also produced another pectinolytic enzyme - pectate lyase (0.01 U/ml). No clear distinction was however, observed in the production profiles of alkaline amylolytic enzymes by the isolates in the two sub-populations [0.20-0.40 U/ml (amylases), 0.24-0.68 U/ml (pullulanases) and 0.01-0.03 U/ml (cyclodextrin glycosyl transferases)]. On the other hand, group 2 isolates were phenotypically identical to one another and also produced similar amounts of proteolytic (0.38, 0.40 U/ml) and amylolytic [amylases (0.06, 0.1 U/ml), pullulanases (0.06, 0.09 U/ml) and cyclodextrin glycosyl transferases (0.01, 0.02 U/ml)] enzymes.Conclusion:The facultatively alkaliphilicB. haloduransand obligately alkaliphilicB. pseudofirmusisolates are attractive biotechnological sources of industrially important alkaline enzymes.