Enzymatic Maceration Research Articles

Comparative Evaluation of Conventional and Emerging Maceration Techniques for Enhancing Bioactive Compounds in Aronia Juice.

Ultrasound and microwave maceration techniques have been utilised to lower production costs and reduce processing time, while also preventing the degradation of nutrients like phenolics and vitamin C and preserving physical properties such as colour and viscosity. In this study, the effects of several traditional (cold, enzymatic, and thermal) and innovative (ultrasonic and microwave) maceration methods on some quality parameters of aronia juice were investigated. Microwave maceration significantly impacted the soluble solids content of the analysed juices and resulted in noticeably darker juice samples compared to the controls, with lower L*/lightness (20.1) and b*/blue-yellowness (-3.2) values and an increased a*/redness value (1.7). Different maceration methods also significantly impacted the rheological properties of the treated juices, among which MW treatment consistently showed a higher viscosity. Sorbitol and fructose were the main sugars identified, while malic acid and quinic acid accounted for 85% of the total acid content. Significant increases in the total sugar and acid concentrations were obtained in the juice samples from ultrasonic, microwave, and enzymatic maceration, while thermomaceration had no significant effect. The concentration of total phenolics ranged from 6.45 g/L in the thermomaceration samples to 9.86 and 14.07 g/L in the ultrasonic and microwave samples, respectively. The obtained results suggest that ultrasonic and microwave technologies were superior in terms of colour improvement and the extraction of sugars, acids, and phenolic compounds compared to traditional maceration methods. Ultrasound and microwave technologies present possible approaches to the improvement of aronia juice production in comparison to traditional methods.

Effect of Heat Pasteurization and Enzymatic Maceration on Yield, Color, Sugars, Organic Acids, and Phenolic Content in the ‘Merlot Kanthus’ Grape Juice

Effect of Heat Pasteurization and Enzymatic Maceration on Yield, Color, Sugars, Organic Acids, and Phenolic Content in the ‘Merlot Kanthus’ Grape Juice

Production of Anthocyanin-Rich Red Rose Petal Extract by Enzymatic Maceration

The use of enzymes to hydrolyze the plant cell matrix is a method known for extracting bioactive substances. The current work used this strategy to produce a rose petal extract rich in anthocyanins that is stable in the presence of marine polysaccharides and has a high antioxidant activity. The process evaluation was carried out sequentially, initially comparing water, ethanol, and their mixtures to anthocyanins extracted in the presence or absence of enzymes. Then, a multi-objective desirability function optimized experimental conditions such as solvent and enzyme concentrations. This study is the first report describing the use of a statistical tool, the central composite rotatable design (CCRD), to optimize anthocyanin extraction from rose petals. This method obtained a maximum extraction of 9.99 mg/g of phenols. The stability of the rose petal extract when using marine polysaccharides retained 60% of the anthocyanins over 28 days without deterioration when protected from sunlight but was practically degraded upon exposure to sunlight. The rose petal extract demonstrated a very high antioxidant capacity of 3.19 μg/mL, close to the literature data for citrus compounds, known to be high in antioxidant compounds for cosmetic food purposes.

Impact of Ultrasound Pretreatment on Juice Yield and Bioactive Content in Juice Produced from Selected Berries Fruit.

The aim of the work was to investigate the effect of ultrasound application during juice pressing. The impact on pressing yield and extraction of bioactive compounds during production of juice from strawberry, blackcurrant and raspberry was evaluated. Juice pressing was carried out after four kinds of mash pretreatment. The tested objects were heated to 55 °C, treated by ultrasound, and/or macerated with an enzyme. The obtained juices were analyzed for the content of bioactive compounds and compared to the 'Control' sample, which was the juice pressed without any pretreatment. Although the results presented here do not conclusively show that enzymatic maceration can be completely replaced by the use of ultrasound, in the case of strawberry and raspberry fruit, juice yield after ultrasound treatment increased almost as much as after enzymatic maceration. Additionally, in the case of raspberry fruit, the antioxidant activity of the juice samples was definitely improved when ultrasound was used. The results from this experiment also showed that it is possible to separate the effect of temperature from the effect of mechanical and chemical actions in ultrasound treatment.

Profile of Phenolic Compounds and Antioxidant Activity of Celery (Apium graveolens) Juices Obtained from Pulp after α-Amylase Treatment from Aspergillus oryzae.

The purpose of this study was to determine the content of certain phenolic compounds, antioxidant activity, pressing efficiency, extract content, and sugars in celeriac juices obtained from the pulp after α-amylase treatment from Aspergillus oryzae. The test material consisted of peeled and unpeeled celery pulp kept at a temperature of 25 °C with and without the enzyme for a period of 30 and 60 min. The juices obtained from them were analyzed for the content of selected phenolic acids and flavonoids using the UPLC-PDA-ESI-MS/MS method, for antioxidant activity measured using the ABTS˙+ and DPPH˙ method, and for the total polyphenol content using the F-C method. Additionally, the juice pressing efficiency, the extract content using the refractometer method, and the sugar content using the HPLC method were checked. Significantly higher antioxidant activity, pressing yield, and average content of caffeic acid glucoside, quinic acid, kaempferol-3,7-di-O-glucoside, and chrysoeriol-7-O-apiosylglucoside were obtained in juices from peeled celery. Maceration of the pulp with amylase resulted in a significant reduction in antioxidant activity compared to control samples. An is-total increase of 17-41% in total flavonoid content was observed in all juices tested after treatment with the enzyme for 30 and 60 min, and the phenolic acid content increased by 4-41% after treatment of the pulp with amylase for 60 min. The 60 min holding of the pulp at 25 °C, including with the enzyme, was shown to decrease the antioxidant activity and the content of quinic acid, ferulic acid, and chrysoriol-7-O-apiose-glucoside in the juices tested compared to the samples held for 30 min, while the content of other phenolic acids and flavonoids increased. In addition, after 60 min of enzymatic maceration, the pressing yield of the juices increased.

Chromosomal characterization mediated by karyomorphological analysis and differential banding pattern in fenugreek (Trigonella foenum-graecum L.): a neglected legume

Fenugreek or Trigonella foenum-graecum L. is a commercially important yet neglected crop of the family Fabaceae, with potent medicinal applications, and can treat several diseases as well. Conventional breeding studies for higher yields of commercial crops largely depend on chromosomal information of the particular species. Despite a number of cytological research being conducted on T. foenum-graecum, a complete characterization of its chromosomes has not been achieved due to the limitations of traditional karyotype analysis methods. A range of chromosomal markers are advantageous to characterize at full extent and identify individual chromosomes rather than relying on only physical metrics. Thus, in this study, in addition to giemsa staining, other approaches like fluorochrome and silver staining were used for the precise karyomorphological analysis of this species. Enzyme maceration and air drying (EMA) based fluorochrome banding with GC-specific stain Chromomycin A3 (CMA), and AT-specific stain 4’,6-diamidino-2-phenylindole (DAPI) applied for the first time for chromosome characterization. The results showed 2n = 16 chromosomes in metaphase cells, with karyotype formula of 2m+6sm. The unique banding pattern observed in the CMA/DAPI and AgNOR staining highlights the AT and GC-rich regions as well as the nucleolar organizer regions (NORs). All this crucial information can further assist in conducting breeding studies of more precision with simultaneously encouraging similar studies that need to be done in other unexploited species of importance.

Sampling and Extraction of Plant Parasitic Nematodes.

The study of nematodes requires availability of nematode specimens and their population densities in plants and soil. This can be achieved using adequate sampling schemes and extraction methods. In this chapter, the most common and suitable sampling and extraction procedures and equipment are described. These include the use of Baermann's funnels, Cobb's decanting and sieving, floating methods such as the Oostenbrink method and Fenwick can, elutriators such as Seinhorst methods, centrifugation methods including that of Coolen, and mechanical and enzymatic maceration. The combination of different methods for cleaning the nematode suspensions is described, such as Cobb's sieving with Baermann's funnels or centrifugation, and for cysts combining Seinhorst's elutriator or Fenwick can with the alcohol methods. Methods for extraction of eggs and/or juveniles of cyst and egg mass forming nematodes, to be used as inoculum or to ascertain egg viability, are also described. Only little information is also noted on the use of molecular tools to identify and quantify nematode populations in soil and roots.

DNA content, ploidy level determination and genetic variations of okra (Abelmoschus esculentus L.) genotypes

Okra (Abelmoschus esculentus L.) is a minor small crop in Jordan; it has attracted a lot of attention as a substitute for conventionalvegetables throughout the world. There are conflicting reports about chromosome numbers in this species. To determine the ploidylevel of different okra genotypes, okra root tips were treated with HCl maceration, enzymatic maceration, and Carmine acid squashing.Treating cells with HCl didn’t macerate the cell in a way that enables chromosome count. The enzymatic treatment combinationshowed no significant effect on cell maceration. Carmine’s acetic acid squashing method was able to digest the cells but in a way thatall chromosomes from neighboring cells gathered, making it difficult to count them from each cell. Flow cytometry as an alternativeway to assess okra ploidy, was considered as an option. The genome size of okra ranged from 4.11 pg 2C in genotype 43 to 6.27 pg 2Cin genotype 30.

A comparison of maceration methods for the preparation of infant skeletal remains for forensic anthropological analysis

Very little literature currently exists prescribing which maceration method to use when preparing infant human remains, resulting in bone quality that is suitable for forensic anthropological analysis. The aim of the study was to test five maceration methods to determine which is most suitable for infant remains for forensic anthropological analysis. The sample included five neonate pig carcasses (Sus scrofa domesticus), ranging between one to three days old. Five maceration methods were tested on the pig carcasses (one pig per maceration method) to determine their effectiveness. The methods included invertebrate maceration by meal worms, chemical maceration by bleach, chemical maceration by borax solution, enzymatic maceration by laundry detergent and sodium carbonate solution, and chemical maceration by sodium hypochlorite. A scoring method was created to assess the effectiveness of each maceration method. Invertebrate maceration and chemical maceration using bleach were the least successful methods of maceration (total maceration score = 8 respectively). Chemical maceration using borax and chemical maceration using sodium hypochlorite achieved complete maceration of the skeletal remains; however, they both resulted in artifacts that are unsuitable for forensic analysis (total maceration score = 14 respectively). Enzymatic maceration using laundry detergent and sodium carbonate was the most successful method (total maceration score = 17). The detergent technique subsequently successfully macerated all five sets of infant human remains. This study has validated that the enzymatic maceration technique using laundry detergent and sodium carbonate can be used to effectively macerate the remains of infant skeletal remains for forensic anthropological analysis.

Comparison of chemical and enzymatic maceration processes for soft tissue removal from arterial silicone casts on skull scaffolds.

The various models used in gross anatomical studies to improve the visualization of blood vessels differ in the amount of manual labour, cost, equipment and time involved. This study aimed to compare chemical and enzymatic maceration processes for soft-tissue removal from arterial silicone casts on skull scaffolds using ringed seal (Pusa hispida) skull specimens. Both processes produced specimens that covered all anatomical aspects required to visualize the intracranial arterial arrangement on a bone scaffold. Overall, the enzyme maceration process was better for production of such specimens, as this process is easy and safe to perform, is less harmful to the bony parts of the specimen, and the resulting specimens are visually more appealing for display and teaching. Compared with previously published models, the end result varied in the amount of dissolved bone tissue and the visual presentation of the model.

Arabinogalactan proteins and polysaccharides compete directly with condensed tannins for saliva proteins influencing astringency perception of Cabernet Sauvignon wines

Wine astringency is thought to be due to salivary protein precipitation; however, the actual mechanism is not well-defined. This study aimed understand the relationship between whole polysaccharide extracts, produced with and without enzyme maceration, and the saliva protein-tannin precipitation reaction. Polysaccharides were analyzed in the context of salivary protein–tannin interactions using gel electrophoresis, quantitative 1H proton nuclear magnetic resonance (qHNMR), size separation chromatography, immunochemistry, and sensory analysis. Polysaccharide addition reduced saliva protein concentration in tannin-saliva protein-polysaccharide mixtures, indicating that native-wine polysaccharides compete with condensed tannins for salivary protein as ligand partners. qHNMR showed that tannin levels were increased by adding polysaccharides, suggesting that in these conditions, polysaccharides interact with saliva proteins via competitive protein-polysaccharide complex formation. Polysaccharides from non-enzyme-treated wines had threshold concentration of 121 mg/mL versus 86 mg/ml for enzyme-treated as detected by a sensory panel. Enzyme-treated polysaccharides changed astringency perception at a lower concentration than non-enzyme-treated polysaccharides.

Maceration Techniques for Human Fetal and Perinatal Bone

Forensic anthropologists working with cases that vary in stages of decomposition are often required to process and macerate remains to complete a forensic analysis. Maceration techniques vary between laboratories, and procedures to facilitate maceration of fetal and perinatal remains are lacking in the literature. This descriptive case study evaluates the use of several maceration techniques for fetal and perinatal remains (n = 2), including cold-water bacterial maceration, hot-water enzymatic maceration, dehydration, and incubation. Dehydration is a new maceration technique previously unpublished. For each technique, the authors assessed ease of maceration, effect on bone quality, and utility for forensic casework and/or donated remains and found all techniques are easy to implement and do not greatly diminish bone quality. Previous research recommends hot-water enzymatic maceration for forensic casework, as it will not degrade DNA and can efficiently remove soft tissues. This case study corroborates this recommendation but finds that incubation may be preferred for fetal remains, as it is quicker and less labor intensive. However, cold-water maceration and dehydration are recommended for donated fetal remains. Cold-water maceration is low maintenance, minimally malodorous, and preferable for disarticulated teaching materials, since this technique avoids any heat-induced warping of fetal bones. Dehydration retains cartilaginous structures and allows for the preservation of articulated elements for comparative specimens in donated collections. By demonstrating several techniques for fetal and perinatal maceration, this case study serves as a starting point toward the creation of general guidelines for forensic anthropology practitioners.

Toward gentle chokeberry juice production by ultrasound-assisted enzymatic maceration

Sustainable processes accompanied by high extraction yields and minimized amounts of by-products are a major goal of current fruit juice production. Controlled degradation of cell wall polysaccharides, in particular pectin, may contribute to reduced emergence of side streams. Possible strategies for the optimization are the selection of enzyme preparations based on comprehensive studies of their activities, the adjustment of maceration temperature toward more gentle conditions, and the application of alternative technologies such as ultrasound (US) during maceration. The present study provides insights into the effects of ultrasound-assisted enzymatic maceration (UAEM) on pectin degradation, total anthocyanin content, thermal and storage stability, and juice yield during chokeberry juice production on pilot-plant scale. The two enzyme preparations applied predominantly possessed polygalacturonase or pectin lyase activity. Cell wall polysaccharide degradation was improved by US and resulted in a 3% increase in juice yield by UAEM using an enzyme preparation that shows mostly polygalacturonase activity. Thermostability of anthocyanins was improved in juices produced using pectin lyase and applying US and matched the stability of anthocyanins in juices produced using polygalacturonase. Storage stability of anthocyanins was improved in juice produced using polygalacturonase during UAEM. UAEM also resulted in lower yields of pomace making the production more resource-efficient. Overall, the use of polygalacturonase has promising potential to advance conventional chokeberry juice production by applying US at gentle conditions.

The effect of enzyme treatment on polyphenol and cell wall polysaccharide extraction from the grape berry and subsequent sensory attributes in Cabernet Sauvignon wines

Pectolytic enzyme maceration is common for producing red wines, but the effects on bitterness and astringency are not well understood. Glycan microarrays assessed polysaccharide diversity and with polyphenol analysis was correlated with sensory data on descriptors of astringency and their perceived levels in enzyme-crafted Cabernet Sauvignon wines. Enzyme use is shown to have no effect on bitterness, but enzyme-macerated wines are more astringent. The data suggests that pectolytic enzymes are much more pronounced in their effect on the cell wall matrix than the ripeness of the berries at harvest and subsequent sensory perception. Enzyme-macerated red wines showed higher levels of polyphenol which were more polymerized and galloylated. The polyphenol-rich wines were described as hard, chalky, grippy, grainy and dry. The non-enzyme wines had elevated levels of arabinogalactan protein and pectin epitopes (notably biomarker mAbs JIM8 and JIM13) with the wines being characterized as soft, fine and velvety.

Study on Clarification of Apple Juice using Enzymes

One of the key challenges in apple juice processing is obtaining a good juice recovery and attaining a juice with good clarity. The presence of pectin and starch components inhibits the juice extraction process and leads to the formation of a cloudy haze which is undesirable in apple juice. For these purposes, maceration enzymes such as pectinase, amylase, cellulose, and hemicellulase are added both before and or after juice extraction to enhance juice recovery percentage and clarify the juice. Process parameters such as type of enzymes used, stage of enzyme addition, incubation time and temperature influence the efficiency of enzymes. The optimisation of these parameters is critically important in apple juice processing for better juice recovery and attaining the desired juice clarity. In this study, juice recovery percentage was compared amongst the control and three treatments- addition of enzyme pectinase at 0.02%, amylase at 0.02% and a combination of both at 0.01%. For the optimisation of process parameters, the type of enzymes used, stage of enzyme addition, incubation temperature, and time were studied as independent variables by comparing with the transmittance value as a dependent variable. Amylase at 0.02%, pectinase at 0.02%, and a combination of 0.01% each of both enzymes were used. The enzyme was added to the pomace and the juice. The treatments were incubated at 200C and 400C. Measurements were done after every 1, 2, 4, and 24 hours of incubation. The juice recovery percentage was not significantly different in the 3 treatments and control where the enzyme was added to the crushed apple. For clarification of apple juice, it is recommended to add the combination of 0.01% each of amylase and pectinase directly to the pomace before juice extraction and incubating the juice obtained at 400C for 24 hours.

On the Effect of Microwave Heating on Quality Characteristics and Functional Properties of Persimmon Juice and Its Residue.

In this study, it was investigated whether integration of microwave-heating into the pretreatment step of persimmon juice processing allows the concomitant production of both functional juice and added-value solid residue from the Diospyros Kaki “Jiro” cultivar. In this direction, persimmon pulp was treated under three different microwave-heating conditions (0.7, 4.2, and 8.4 kJ/g) prior to enzymatic maceration and compared to the non-heated material. Irrespective of microwave energy employed, the proposed hybrid treatment was highly efficient in terms of juice yield (70% w/w). The mildest heating conditions resulted in juice and residue that were both of inferior quality. Intensification of the microwave energy reduced the microbial load of the juice up to 2-log without compromising the content in total soluble solids, sugars, and L-ascorbic acid. Under the most drastic conditions, the juice was enriched in gallic acid, polyphenols, and potent DPPH● scavengers, but its orange color faded and was more acidic. In parallel, the solid juice residue retained pro-vitamin A carotenoids (~278 µg retinol activity equivalents) and low-methoxy pectin (9 g/100 g DW). Overall, our findings can assist the efforts of the local juice processing industry to utilize persimmon fruits through energy-efficient technologies in a sustainable approach.

Process optimization in the obtention of microfiltered banana ( Musa cavendish ) juice by response surface methodology

This study is focused on the optimization of the clarification process of banana juice (Musa cavendish). Two central composite rotational designs (CCRD) were elaborated that aim at the enzymatic maceration stage: having (first design) independent variables such as pectinase, cellulase, and amylase concentrations, and (second design) time and amylase concentration. The dependent variables of both designs were the permeate flow (Jp) and apparent viscosity (µ) of the pulp, at a significance level of p < .10. The results indicated a negative effect in relation to cellulase and optimum concentration of 500 mg/L of pectinase. The second design achieved optimal conditions at a 1 hr maceration time and amylase concentration of 1,000 mg/L. The resistances associated with fouling (reversible + irreversible) were predominant in both operations. It is concluded that there is a technical feasibility in using combined methods to obtain clarified banana juice, allowing the scale to be extended from pilot to industrial. Practical applications The clarified banana juice is a versatile beverage that can be used in ready-to-eat formulations or as an ingredient in other products, but in order to make the production of this clarified juice feasible, it is necessary to improve the microfiltration step. Due to the high viscosity of crude banana pulp, the current main process used to allow the microfiltration is a dilution step, which turns future industrial applications unfeasible. In this study, a combined method of enzymatic maceration and microfiltration was optimized, achieving a technical viability for industrial purposes, also, a complementary study of fouling in microfiltration was included, with the objective to contribute to scientific and industrial communities regarding the production of clarified fruit juices.

Karyotype diversity in cultivated and wild Indian rice through EMA-based chromosome analysis

India is known for its diverse cultivated and wild rice germplasm. In today's crop improvement programmes, wild relatives are much-needed genetic repository of valuable traits. Analysis of genetic diversity at the chromosomal level is one cost-effective tool to unlock foundational information related to genetics and plant breeding. Presently, enzymatic maceration and air-drying method (EMA) has been applied for the first time in six cultivated and nine wild Indian rice (diploid and tetraploid). EMA method following Giemsa staining has yielded large numbers of cytoplasm free metaphase plates with distinct chromosome morphology. Detailed analysis has revealed karyotype diversities in terms of total chromatin length (TCL), chromosome morphology and location of sat chromosomes within and between the studied species. Most of the cultivated rice has gained additional amount in TCL during the period of domestication in comparison to their progenitor Oryza nivara. Morphological clarity of the small chromosomes of rice was much required and has helped to identify individual chromosomes in the diverse karyotypes. Diversity in landmark SAT chromosomes is another important observation, not reported previously in Indian rice. Present study has shown that in most of the O. sativa members, the 10th pair contains SAT except one where 6th pair is satellited. On the other hand, diversity of SAT in diploid and tetraplod wild species has been recorded on 5th, 7th and 8th chromosome pairs and on 9th, 12th, 22nd and 23rd chromosome pairs, respectively. Karyomorphometric indices has helped to construct dendrogram to elucidate intraspecies and interspecies relationships. Untapped genetic diversity recorded in Indian rice through chromosomal analysis will be useful to the breeders and genome researchers.

Effects of ultrasound on the enzymatic degradation of pectin

Ultrasound-assisted enzymatic maceration (UAEM) has gained considerable interest in the fruit juice industry, owing to its potential to increase juice yield and content of polyphenols while simultaneously saving time and energy. In this study, the effects of UAEM (ultrasonic probe, 20 kHz, 21 W*cm−2 and 33 W*cm−2) on pectin degradation in a continuous circulation system were investigated over 60 and 90 min. Main pectinolytic enzymes activities of (polygalacturonase, pectin lyase and pectin methylesterase) of a commercial enzyme preparation were examined for individual synergistic effects with US. Pectin hydrolysis by UAEM differed significantly compared to treatment with ultrasound or enzymes alone regarding the profile of degradation products compared to treatment with ultrasound or enzymes alone. Ultrasound fragmented pectin to less branched oligomers of medium molecular weight (Mp approx. 150 kDa), which were further degraded by pectinolytic activities. The low molecular weight fraction (<30 kDa), which is known to be beneficial for juice-quality by adding nutritional value and stabilizing polyphenols, was enriched in small oligomers of homogalacturonan-derived, rhamnogalacturonan I-derived, and rhamnogalacturonan II-derived residues. Synergistic effects of ultrasound application enhanced the effective activities of polygalacturonase and pectin lyase and even prolonged their performance over 90 min, whereas the effective activity of pectin methylesterase was not affected. Final marker concentrations determined by each enzyme assay revealed a considerable higher total process output after UAEM treatment at reduced temperature (30 °C) comparable to the output after conventional batch maceration at 50 °C. The obtained results demonstrate the high potential of UAEM to produce high-quality juice by controlling pectin degradation while reducing process temperature and equally highlight the matrix and enzyme specific effects of a simultaneous US treatment.

Enzymatic maceration and liquefaction of pumpkin ( Cucurbita moschata L.) flesh for the preparation of a suitable base feed for spray drying

Enzymatic maceration and liquefaction of pumpkin ( <i>Cucurbita moschata</i> L.) flesh for the preparation of a suitable base feed for spray drying