Production Of Pectic Oligosaccharides Research Articles

Towards a sustainable circular bioprocess: Pectic oligosaccharides (POS) enzymatic production using passion fruit peels

A bioprocess was designed using passion fruit peels for the enzymatic production of POS. For this purpose, three commercial pectinase complexes were tested, and one was selected based on the analysis of viscosity change and galacturonic acid release during the reaction. Operating conditions were further determined at laboratory and bench scale, including time and enzyme dosage. Subsequently, the bioprocess was validated at pilot scale in a 40 L reactor, obtaining a highly concentrated syrup with 73.3 g POS per 100 g total solids, and 22.6 g total POS per 100 g passion fruit peel processed. Thus, the technical feasibility of a bioprocess leading to the production of a potential prebiotic syrup was proven, contributing to the SDG 12-3, “reduce food losses along production and supply chains”. These findings highlight for the first time the utilization of passion fruit peels in creating a prebiotic POS syrup through an enzymatic process involving commercial complexes. The straightforward nature of the bioprocess, comprising just four unitary operations, combined with the utilization of agricultural waste and commercial enzymes, presents compelling advantages. Particularly appealing to food processors aiming to enhance the circularity and sustainability of their operations while reducing reliance on external prebiotic ingredients.

Innovative biocatalyst synthesis of pectinolytic enzymes by cross-linking strategy: Potentially immobilised pectinases for the production of pectic-oligosaccharides from pectin

Pectinases are outstanding multienzymes, which have the potential to produce new emerging pectic-oligosaccharides (POS) via enzymatic hydrolysis of pectin. However, free pectinase is unable to undergo repeated reaction for the production of POS. This study proposed a sustainable biocatalyst of pectinases known as cross-linked pectinase aggregates (CLPA). Pectinase from Aspergillus aculeatus was successfully precipitated using 2 mg/mL pectinase and 60 % acetone for 20 min at 20 °C, which remained 36.3 % of its initial activity. The prepared CLPA showed the highest activity recovery (85.0 %), under the optimised conditions (0.3 % (v/v) starch and glutaraldehyde mixture (St/Ga), 1.5: 1 of St/Ga, 25 °C, 1.5 h). Furthermore, pectin-degrading enzymes from various sources were used to produce different CLPA. The alteration of pectinase secondary structure gave high stability in acidic condition (pH 4), thermostability, deactivation energy and half-life, and improved storage stability at 4 °C for 30 days. Similarly to their free counterpart, the CLPA exhibited comparable enzymatic reaction kinetics and could be reused eight times with approximately 20 % of its initial activity. The developed CLPA does not only efficaciously produced POS from pectin as their free form, but also exhibited better operational stability and reusability, making it more suitable for POS production.

Continuous hydrolysis of mango peel pectin for the production of antibacterial pectic oligosaccharides in packed-bed reactor using immobilized polygalacturonase

The aim of this study was to continuously hydrolyze mango peel pectin, an agricultural waste, into pectic oligosaccharides (POS) with antibacterial activity. A packed-bed reactor (PBR) with immobilized polygalacturonase was designed, and the effects of the process conditions (flow rate, substrate concentration, and enzyme amount) were evaluated. The optimal conditions were obtained using response surface analysis, and a stable POS production with a yield of 94.56% was achieved. The PBR retained 82% of its initial activity after continuous operation for 72 h. The POS products exhibited good antibacterial activity against E. coli, S. aureus, B. subtilis, and S. typhimurium. The destructive effects of POS on the membrane system integrity were observed via scanning electron microscopy and the leakage of intracellular nucleic acids. Thus, the PBR is a powerful tool for continuous POS production, and hence, it can be applied as an alternative to chemical preservatives in the food industry.

Production of pectic-oligosaccharides from pomelo peel pectin by oxidative degradation with hydrogen peroxide

Oxidative depolymerization of alkali- and acid-extracted pomelo pectins was performed using 1% hydrogen peroxide (H2O2) with a microwave power of 550 W for 10 min. Pectic-oligosaccharides (POS) produced from the acid-extracted methyl-esterified pectin contained higher amounts of DP1 and DP2 than that from the nearly ester-free alkali-extracted pectin, and the loss of these small-size products during recovery resulted in a lower POS yield (25.0%) compared to the alkali-extracted pectin (57.7%). Degradation of the alkali-extracted pectin with 3 and 5% H2O2 led to a decrease in precipitable POS yield. The relative amount of large-sized POS decreased as the H2O2 concentration increased. An increase in the microwave power to 1100 W had no significant effect on overall yield, but the average size shifted to be lower. The results of sugar composition and identification of the degraded products with ESI-MS confirmed the existence of several POS species with different sizes and structures.

Agro waste derived pectin poly and oligosaccharides: Synthesis and functional characterization

Humans consume majorly the plants derived carbohydrates in their diet. The major component of plant material comprises of complex polysaccharides rich in arabinan, arabinogalactan, rhamnogalactan and polygalacturonic acid. Dietary fibers are resistant to human digestive enzymes and are exclusively fermented by the beneficial bacteria present in the intestine, termed as microbial accessible carbohydrates (MAC). The beneficial bacteria utilize the MAC to produce antimicrobial substances and short chain fatty acids to maintain the complex microbial diversity of gut and fight against pathogenic bacteria in the intestine. Currently, available prebiotics in the market are limited and some of them are digested in the proximal part of intestine. As a result, the demand for new prebiotic ingredients with improved functional properties is increasing tremendously. Pectin and pectic oligosaccharides (POS) are resistance to human digestive enzymes and are fermented by bacteria in the intestine and subsequently enhance the gut health. The present review describes about the various production, purification and functional characterization of pectin and pectin derived oligosaccharides from agro waste. Various current and futuristic functional properties of pectin and POS has been explained. This study will help the food processing industry to develop several plants derived futuristic functional food products. • Production and purification of pectin and POS from different agro waste is reviewed. • Current application of pectin in food, and anti-cancer agent is discussed. • Role of pectin as nanocomposite for various biomedical application is reviewed. • Anti-microbial and prebiotic properties of pectin and POS is described.

Immobilization of polygalacturonase for the preparation of pectic oligosaccharides from mango peel wastes and assessment of their antibacterial activities

In this study, we immobilized polygalacturonase by sol-gel entrapment for the continuous preparation of pectic oligosaccharides (POS) from mango peel wastes and evaluated their antibacterial activity. We characterized the immobilized polygalacturonase by scanning electron microscopy, thermo gravimetric analyzer, fourier transform infrared spectrometer and raman spectrum. Results showed that the polygalacturonase was successfully immobilized by the sol-gel encapsulation, which retained as much as 94.6% of its initial activity. The immobilized enzyme exhibited enhanced pH tolerance and thermal stability compared with its free counterpart. For example, the immobilized enzyme retained 57% of relative activity at 55 °C for 2 h, whereas the free enzyme retained only 17% of relative activity. Moreover, the immobilized enzyme showed good recyclability, which retained 65% of its initial activity after 6 batch reactions. Furthermore, we examined the antibacterial activities of POS prepared by the immobilized polygalacturonase hydrolysis by typical bacteria. The POS was mainly polygalacturonic acid formed by →1)-α-GalpA-(4→ and possessed the best antibacterial activity with a polymerization degree of 2–4. Therefore, we found it could be used as a natural antibacterial agent in food industry. These results suggested that immobilized polygalacturonase was efficient in terms of its high catalytic activity and stability, and it can be applied to continuous POS production.

Continuous production of pectic oligosaccharides from onion skins with an enzyme membrane reactor

The aim of this research was to valorize onion skins, an under-utilized agricultural by-product, into pectic oligosaccharides (POS), compounds with potential health benefits. To achieve high hydrolysis performance with the multi-activity enzyme Viscozyme L, an innovative approach was investigated based on a cross-flow continuous membrane enzyme bioreactor (EMR). The influence of the various process conditions (residence time, enzyme concentration, substrate concentration) was investigated on productivity and yield. The composition of the POS mixtures in terms of mono- and oligosaccharides was assessed at the molecular level. At optimized conditions, a stable POS production with 22.0g/L/h volumetric productivity and 4.5g/g POS/monosaccharides was achieved. Compared to previous results obtained in batch for the enzyme Viscozyme L, EMR provided a 3–5× higher volumetric productivity for the smallest POS. Moreover, it gave competitive results even when compared to batch production with a pure endo-galacturonase enzyme, demonstrating its feasibility for efficient POS production.

A Novel Non-Cumbersome Approach Towards Biosynthesis of Pectic-Oligosaccharides by Non-Aflatoxigenic Aspergillus sp. Section Flavi Strain EGY1 DSM 101520 through Citrus Pectin Fermentation.

Pectic-Oligosaccharides (POS) have a growing potential in food and feed industries. To satisfy the demand of worldwide markets from POS and avoid the shortcomings of currently applied methodologies encountered in their preparation, the present study highlights a novel robust approach for POS biosynthesis. In the current approach, Aspergillus sp.section Flavi strain EGY1 DSM 101520 was grown on citrus pectin-based medium as a core POS production medium. POS' levels accumulated in the fungal fermentation broth were optimized through a three step sequential statistical mathematical methodology; Plackett-Burman design (PBD), Box-Behnken design (BBD) and canonical analysis. Three key determinants namely citrus pectin, peptone and NaH2PO4 were pointed out by PBD to impose significant consequences (P<0.05) on the process outcome (POS' levels). Optimal levels of these key determinants along with maximal of POS' levels were set by BBD and canonical analysis to be 2.28% (w/v) citrus pectin, 0.026% (w/v) peptone and 0.28% (w/v) NaH2PO4 to achieve a net amount of 1.3 g POS /2.28 g citrus pectin. Through this approach, a yield of 57% (w/w) POS of the total citrus pectin was obtained after 24 h of fungal growth on optimized citrus pectin–based medium. A fold enhancement of 13 times in POS' levels released in the fermentation fungal broth was realized by the end of the optimization strategy. This novel robust approach is considered a new insight towards POS biosynthesis via efficient, rapid and non-cumbersome procedure. To the best of authors' knowledge, the present work is the first article underlining detailed POS production from the fermentation broth of a fungus growing on citrus pectin-based medium.

Enzymatic production of pectic oligosaccharides from onion skins

Onion skins are evaluated as a new raw material for the enzymatic production of pectic oligosaccharides (POS) with a targeted degree of polymerization (DP). The process is based on a two-stage process consisting of a chelator-based crude pectin extraction followed by a controlled enzymatic hydrolysis. Treatment of the extracted crude onion skin’s pectin with various enzymes (EPG-M2, Viscozyme and Pectinase) shows that EPG-M2 is the most appropriate enzyme for tailored POS production. The experiments reveal that the highest amount of DP2 and DP3 is obtained at a time scale of 75–90min with an EPG-M2 concentration of 26IU/mL. At these conditions the production amounts 2.5–3.0% (w/w) d.m for DP2 and 5.5–5.6% (w/w) d.m for DP3 respectively. In contrast, maximum DP4 production of 5.2–5.5% (w/w) d.m. is obtained with 5.2IU/mL at a time scale of 15–30min. Detailed LC–MS analysis reveals the presence of more methylated oligomers compared to acetylated forms in the digests.

Conditions of limited hydrolysis producing pectic oligosaccharides (pos) of high concentration passion fruit pectin

Pectic oligosaccharides (POS) have been proposed as a new class prebiotics capable of exerting a number of health-promoting effects. POS can be obtained by limited hydrolysis enzyme of pectin. The present study investigates the conditions of limited hydrolysis passion fruit pectin (Passiflora edulis flavicarpa) by pectinase. Pectin with concentration 1-5% (w/v) was hydrolyzed by Pectinex Ultra SP-L 30-50 U/g, pH 3-5, 40-60°C at 200-350 rpm with hydrolysis time of 1-7 hours. Research results show that pectin 3% (w/v), 40 U/g, pH 4, 50oC, 250 rpm, products have 18.76 mg POS/ml. Response surface methodology was used to model the processes. The temperature (45-55°C), enzyme concentration (30-50 U/g), and speed agitation (200-300 rpm) were the factors studied. Operational conditions leading to a maximum POS production were calculated from models being 53oC, 44 U/g, 270 rpm. Under the conditions, the model predicted that 24.55 mg/ml of POS can be obtained from pectin. Conversions to product increased from 62,53 to 81,83%. Products contain di and tri galacturonic acid.

Pectic oligosaccharides production from orange peel waste by enzymatic hydrolysis

SummaryOrange peel waste (OPW) was evaluated as a raw material for the enzymatic production of pectic oligosaccharides. In a preliminary step, free sugars and other water‐soluble compounds were extracted from OPW to obtain a substrate for enzymatic hydrolysis (EHS), to achieve two objectives: (i) removal of sugars and impurities, and (ii) production of a sugar‐rich stream suitable for further utilisation, according to the ‘biorefinery’ philosophy. EHS was treated with hydrolytic enzymes (pectinases and cellulases), and the effects of selected operational variables (enzyme charges and reaction time) on selected dependent variables (measuring the conversions of polysaccharides into mono‐ and oligosaccharides, and liquor recovery) were assessed. Mathematical models were developed for this purpose. Under selected conditions, the models predicted that 100 kg of EHS yielded 7.5 kg of gluco‐oligosaccharides, 4.5 kg of galacto‐oligosaccharides, 6.3 kg of arabino‐oligosaccharides and 13 kg of oligogalacturonides (contained in 991.6 kg of liquor).