Pectinase Production Research Articles

Halophilic Pectinase-Producing Bacteria from Arthrocnemum macrostachyum Rhizosphere: Potential for Fruit–Vegetable Juice Processing

This study aimed to isolate salt-tolerant pectinolytic bacteria from the rhizosphere of a salt marsh plant and utilize their pectinases for the clarification of detox juice preparation. Sixteen halophilic bacterial strains were isolated from the rhizospheric soil of Arthrocnemum macrostachyum. The isolates were screened for pectinase activity, and two strains, ASA21 and ASA29, exhibited the highest pectinase production in the presence of 2.5% NaCl, reaching 13.3 and 14.1 IU mL−1, respectively. The strains were identified as Bacillus paralicheniformis and Paenibacillus sp. by 16S rDNA sequencing and phylogenetic analysis. Growth kinetics and pectinase production studies revealed that both strains produced pectinase during the log phase, with ASA29 demonstrating higher growth and pectinase titers. The pectinase from ASA29 exhibited enhanced activity in the presence of 3% NaCl. The pectinases from both strains were applied for the clarification of detox juice prepared from beetroot, carrots, and apples. The use of 20 IU mL−1 pectinase from ASA29 for 2–3 h yielded > 96% juice with high total phenolic content and antioxidant activities. This study highlights the potential of salt-tolerant pectinolytic bacteria from the rhizosphere for biotechnological applications, particularly in the clarification of juices with high salt content.

Genetic Diversity and Functional Potential of Streptomyces spp. Isolated from Pachmarhi Biosphere Reserve, India.

Streptomyces is a diverse genus, well known for producing a wide array of metabolites that have significant industrial utilization. The present study investigates the genetic and functional diversity of Streptomyces spp. isolated from the Pachmarhi Biosphere Reserve (PBR), India, an unexplored site. The 16S rRNA gene sequencing and analysis revealed 96 isolates belonging to 40 different species indicating a substantial phylogenetic diversity. The strains were clustered into two groups: a major cluster with 94 strains and a small cluster with two strains. BOX- PCR analyses revealed an incredible genetic diversity existing among the strains of Streptomyces spp. in PBR. The analyses revealed the intra-species diversity and inter-species closeness within the genus Streptomyces in the study area. Qualitative screening for enzyme production has shown that 53, 42, 41, 11, and 54 strains tested positive for CMCase, xylanase, amylase, pectinase, and β-glucosidase, respectively. Additionally, 54 strains tested positive for PHB production. The strains were assayed quantitatively for the production of CMCase, xylanase, amylase, and pectinase. Streptomyces sp. MP9-2, Streptomyces sp. MP10-11, Streptomyces sp. MP10-18, and Streptomyces sp. MP10-6 recorded maximum CMCase (0.604 U/mL), xylanase (0.553 U/mL), amylase (1.714 U/mL), and pectinase (13.15 U/mL) activities, respectively. Furthermore, several strains demonstrated plant growth-promoting traits, viz. zinc and phosphate solubilization and production of ammonia, HCN (hydrogen cyanide), and IAA (Indole acetic acid), and nitrogen fixation. Fifty strains showed antifungal activity against Fusarium oxysporum f. sp. lycopersici with inhibitions ranging from 7.5 to 47.5%. Current findings underscore the ecological and biotechnological significance of Streptomyces spp. in the unexplored habitat of PBR.

Valorization of agro-industrial waste for pectinase production and its influence on circular economy

Pectinases are a diverse group of enzymes that play a crucial role in modifying or breaking down complex pectic substances. Pectinases are widely distributed among bacteria, fungi, and plants. The global demand for microbial pectinase has significantly increased due to its broad applicability and efficient catalytic capabilities across multiple industries including food processing, textiles, and biofuel production. Their commercial production often relies on expensive substrates, contributing to economic inefficiency and environmental burdens. Utilizing agro-industrial waste and microorganisms for pectinase production offers a rational solution to two interconnected challenges: the cost-effectiveness of enzyme production and the environmental impact of waste generation. Moreover, the valorization of waste materials not only contributes to efficient enzyme production but also exemplifies a circular approach by minimizing environmental impact and promoting sustainable resource efficiency to bioprocessing. This review offers a thorough examination of microbial pectinases, including their production from agro-industrial waste, their various industrial applications, and the current market landscape. It also delves into recent advancements in enzyme development and optimization techniques that have significantly boosted the efficiency and cost-effectiveness of pectinase production. By highlighting these developments, the review emphasizes the potential for this approach to enhance industrial practices and contribute to environmental sustainability.

Experimental study and mathematical modeling of the pectinases production by Aspergillus flavipes FP-500 in an airlift bioreactor

Mathematical models are indispensable for designing, optimizing, and controlling large-scale bioprocesses. In the present work, the production of pectinases was studied experimentally using an internal loop airlift bioreactor, pectin as a substrate, and Aspergillus flavipes FP-500 as a biocatalyst. The N-tanks-in-series (NTIS) model was implemented to predict the behavior of fungal growth, pectin and dissolved oxygen consumption, pectinases production, and the oxygen mass transfer rate from gas phase to liquid culture medium. A double Monod-Logistic kinetic model was used to describe the biomass growth rate as a function of biomass, pectin, and dissolved oxygen concentrations. In contrast, a Luedeking-Piret kinetic model was used to describe the production rate of endo and exo pectinases. A hydrodynamic model was utilized to estimate gas hold-ups, volumetric mass transfer coefficients, and air inflow velocities. Good agreement was observed between the experimental data and the theoretical results, demonstrating the predictive capacity of the NTIS model to describe pectinase production, the oxygen consumption rate, and the oxygen evolution in the gas phase. The model highlighted its robust capability to capture the critical parameters of aerobic fermentation processes. Therefore, it could be used as a tool for the scalability of the airlift bioreactors.

Microbial Multienzyme Viz., Pectinase, Cellulase and Amylase Production Using Fruit and Vegetable Waste as Substrate—A Review

Around 70 million metric tonnes of fruit and vegetable waste (FVW) are produced each year and are eventually discarded as wholesale garbage. Microorganisms decompose this FVW, which has led to environmental contamination, greenhouse gas emissions, and other impacts related to climate change. If FVW are used properly, they can reduce environmental damage and also boost a nation’s economy. FVW contain vast amounts of biopolymers, viz., pectin, cellulose, and starch, all of which are hydrolysed by microbes with the aid of the pectinase, cellulase, and amylase enzymes, respectively. Therefore, in light of this, the intervention of microorganisms for the production of pectinase, cellulase, and amylase could be a safe, cost-effective, and eco-friendly approach for the precise utilisation of FVW. Nowadays, thermophilic multienzymes are extracted from a group of hot spring microbes. Thermophilic multienzymes are more capable of surviving at high temperatures and have less degrading capability. Moreover, through this advancement, we can obtain vast amounts of pectinase, cellulase, and amylase enzymes within a short period of time. This microbial enzyme preparation might be helpful in food, textiles, paper, pulp, animal feed supplements, detergents, juice/pulp clarity, leather, and other related sectors.

Isolation and Characterization of Aeromonas taiwanensis Strain for Simultaneous Production of Cellulase, Amylase, Pectinase, and Protease Enzymes

Isolation and Characterization of Aeromonas taiwanensis Strain for Simultaneous Production of Cellulase, Amylase, Pectinase, and Protease Enzymes

Optimization of Culture Conditions and Characterization of Pectinase from Isolated Bacteria and Application in Juice Clarificationz`

Microbial pectinase has high demand in different industries. Pectinase producing bacteria were isolated and purified from soil samples and screened by primary and secondary screening for pectinase production. Out of the 13 bacterial samples, sample 4.1(S4.1) showed consistent enzyme activity. Optimum production of pectinase was high when incubated at 24-hour at 37°C with 2% substrate concentration maintaining pH6. Optimum pectinase activity was observed at temperature 37°C and pH 6. Cu++ was found as activator and Zn++, Ca++ as inhibitors of pectinases. The values for Km and Vmax were found to be 1.181mg/ml and 471.69µM/min respectively. Pectinase was efficiently partially purified by 90% acetone saturation. Specific enzyme activity for partially purified enzyme with acetone was 96870 U/mg and purification fold was 35.41. The utilization of low-cost agro-industrial wastes as substrates has been preferable in pectinase production. The sweet lemon and lemon fruit peel powder was observed good substrate for pectinase production. Furthermore, the yield and clarity of sweet lemon juice increased as the concentration of pectinase increased, indicating its potential use in juice processing. Experimental results suggest pectinases from S4.1 was of high value and can be utilized for fruit juice clarification.

Study of the Pectinase Production from Soursop and Cherimoya Pulp for Agro-Industrial Waste Reduction in Colombia

Pectic enzymes are an eco-friendly alternative to improve extraction processes for the food industry. The objective of the research was to obtain pectinase from the pulp of soursop (Annona muricata L.) and cherimoya (Annona cherimola L.), as an alternative to reduce agro-industrial waste in Colombia; involving the study of the factors influencing the protein content of the enzymes extracted from different stages and types of fruits, the conditions for enzymatic activity, and the potential application in the food industry, particularly in extraction processes and juice clarification. A factorial design 23, spectrometric and potentiometric techniques evidenced that the protein content depends on factors such as fruit stage (suitable “s” and unsuitable “u” fruits for consumption) and fruit type (soursop and cherimoya). The higher protein content was obtained by the precipitation method in comparison to ion exchange chromatography. Cherimoya fruit presented a higher protein content of 326 ± 26 mg by precipitation, however, the soursop samples evidenced higher enzymatic activity, 2.2 ± 0.1 U/mL. The pectinase obtained was an acidic enzyme of type pectin methylesterase with an enzymatic activity of 27.0 ± 0.9 U/mL, under the optimal conditions of pH 4 and temperature of 40 °C. The application in blackberry juice evidenced a decrease in the absorbance and pH changes. From the current research, the pulp of soursop and cherimoya was proven as a cheap source to obtain pectinase with enzymatic activity for juice clarification, and this process is an alternative to reduce the negative impact on the environment and an economical source for the local producers.

Pectinases production using an exotic caatinga passion fruit waste and wheat bran as a substrate for clarification of grape juice

Caatinga passion fruit waste (CPFW):Wheat bran has shown reliable performance for enzyme production. However, the mixture of wheat bran and CPFW may provide all nutrient sources needed by Aspergillus niger for pectinase generation. This work investigates the production of pectinolytic enzymes using a substrate composed of CPFW:Wheat bran using Aspergillus niger FSDE16 via semi-solid-state fermentation (SSSF). A blend of wheat bran:CPFW (1:1, g/g) with 60% moisture content was inoculated with 106 spores/mL for 144 h. Substrates were fermented using the 7 combinations of temperature (25–75 °C) and pH (3–6) dispose in an experimental design. Upon the completion of fermentation, all analyses were conducted including the enzymatic production, optimal activity, and enzymatic thermostability. Afterwards, grape juice clarification was conducted, and the clarified juice was evaluated considering viscosity reduction, yield increase, turbidity reduction, yield increase, colour, and phenolic profile. Results demonstrate that enzyme stability was achieved within the temperature range of 30–40 °C, with optimal activity recorded at 50 °C (42.7 U/g). The application of crude enzymatic extract in grape juice clarification led to an increase of yield (>280%), and a viscosity (>80%) and turbidity (>98%) reduction. The clarification treatment revealed a significant increase in the phenolic compounds evaluated, except epicatechin gallate, Pro-B2, and resveratrol, which reduced their concentrations. The clarification resulted in a colour intensity reduction. This study underscores the potential of using food waste for enzyme production, presenting a sustainable approach with implications for cost reduction and waste management in the food industry.

Optimization of conditions for production of pectinase in solid-state fermentation with Aspergillus flavus using dried Assam lemon peel powder as substrate

The culture conditions for pectinase production with Aspergillus flavus by solid-state fermentation (SSF) using dried Assam lemon peel were optimized and the pectinase was used for clarification of grape juice. A central composite rotatable design (CCRD) was used for optimization experiments with moisture content (50–90 %), incubation temperature (20–60 ℃), incubation time (96 h–192 h), and urea concentration (0–1 %) as independent variables, and pectinase activity as dependent variable. The optimum conditions for pectinase production obtained were moisture content of 60 %, incubation temperature of 50 ℃, incubation time of 96 h, and urea concentration of 0.30 % with a pectinase activity of 3.68 ± 0.20 U/mL where one unit of pectinase activity corresponds to number of µmole of galacturonic acid released in one minute in 1 % pectin solution at 37 °C and 4.8 pH. The crude pectinase was found to have maximum activity at a temperature of 40 ℃ and pH 4 and had an apparent Vmax and Km of 6.10 U/mL and 6.62 mg/mL respectively. The grape juice clarified using the crude pectinase showed decreased optical density (1.45 ± 0.06 to 1.17 ± 0.07) and viscosity (1.76 ± 0.10 to 1.46 ± 0.07 mPa.s) and increased reducing sugar content (13.90 ± 0.10 to 16.96 ± 0.06 g/100 mL juice), and total soluble solids (17.77 ± 0.16 to 18.50 ± 0.30 °Brix).

Description of a new genus of the Pectobacteriaceae family isolated from water in coastal brackish wetlands of the French Camargue region, Prodigiosinella gen. nov., including the new species Prodigiosinella aquatilis sp. nov

The Pectobacteriaceae family comprises plant pathogens able to provoke diverse diseases, including plant maceration due to the production of pectinases disrupting the plant cell wall. To better understand their diversity, a survey of pectinolytic bacteria was performed in brackish lakes of the French region La Camargue near the Mediterranean Sea. The genome of six atypical isolates was sequenced; their size is around 4.8 to 5.0 Mb, including a plasmid of 59 to 61 kb; their G+C values range from 49.1 to 49.3 mol%. Phylogenetic analyses indicated that the novel strains form a new clade of Pectobacteriaceae that branches at the basis of the group encompassing the genera Lonsdalea, Musicola, and Dickeya. Based on phenotypic, genomic and phylogenetic characteristics, we propose the creation of a new genus with the name Prodigiosinella gen. nov. Both the phenotypic and phylogenetic analyses separated the strains into two distinct subgroups, G1 and G2. The type strain LS101T (CFBP 8826T = LMG 32072T) and strain CE70 (CFBP 9054 = LMG 32867) are representative G1 and G2 members, respectively. Three genomic methods were used to analyze DNA–DNA relatedness: digital DNA–DNA hybridization (isDDH), average nucleotide identity (ANI), and genome alignment fraction (AF). They revealed a close relationship between genomes of the two groups, supporting their appurtenance to a same species for which we propose the name Prodigiosinella aquatilis sp. nov. Four strains previously designated as Serratia sp. (ATCC 39006), Brenneria “ulupoensis” (K61) or Erwinia sp. (MK01 and MK09) belong to the new genus Prodigiosinella.

Production and immobilization of pectinases from Penicillium crustosum in magnetic core-shell nanostructures for juice clarification

Global market of food enzymes is held by pectinases, mostly sourced from filamentous fungi via submerged fermentation. Given the one-time use nature of enzymes to clarify juices and wines, there is a crucial need to explore alternatives for enzyme immobilization, enabling their reuse in food applications. In this research, an isolated fungal strain (Penicillium crustosum OR889307) was evaluated as a new potential pectinase producer in submerged fermentation. Additionally, the enzyme was immobilized in magnetic core-shell nanostructures for juice clarification. Findings revealed that Penicillium crustosum exhibited enzymatic activities higher than other Penicillium species, and pectinase production was enhanced with lemon peel as a cosubstrate in submerged fermentation. The enzyme production (548.93 U/mL) was optimized by response surface methodology, determining the optimal conditions at 35 °C and pH 6.0. Subsequently, the enzyme was covalently immobilized on synthesized magnetic core-shell nanoparticles. The immobilized enzyme exhibited superior stability at higher temperatures (50 °C) and acidic conditions (pH 4.5). Finally, the immobilized pectinases decreased 30 % the orange juice turbidity and maintained 84 % of the enzymatic activity after five consecutive cycles. In conclusion, Penicillium crustosum is a proven pectinase producer and these enzymes immobilized on functionalized nanoparticles improve the stability and reusability of pectinase for juice clarification.

The Pectinase Production Potential of Microorganisms Isolated from Agricultural Soil and Rotten Fruits in Shiraz, Southwest of Iran

The Pectinase Production Potential of Microorganisms Isolated from Agricultural Soil and Rotten Fruits in Shiraz, Southwest of Iran

Gene Modulation Effect on Phyto-Compound Expression

Biocatalyst hydrolytic function, socio-economic importance and industrial relevance were identified in this investigation and the aim of the research was to determine effect of enzyme infusion on phyto-compounds yield and antimicrobial expression. The following activities were carried out to achieve the objective: i. construct of a hyperactive Bacillus subtilis sub specie using a modulation kit, ii. production of amylase, cellulase and pectinase using the successfully modulated strain, iii. Applying a consortium of the enzymes to infuse powdery samples of Calotropis procera Ait Leaf under synthase proximate conditions of pH 6 at 60oC for 1 hours, and iv. analysis on extract to ascertain phyto-compund yield and antimicrobial expression. The following results were recorded: i. infused extract with a consortium of catalysts showed high impact of 35% increment in secretion of terpenoids, flavonoids, anthraquinone, alkaloids, steroids, tannins, and saponins that were analysed and identified to be present compared to extracts of dimethyl ether, 70% ethanol, and methonal; ii. results also recorded high impact of boioactive agent expression against test strains with the following phyto-restraint articulations of 33.3mm, 28.6 mm, 30.4 mm, 31.6 mmm, 30.6 mm, and 22.3 mm for Escherichia coli, Enterobacter aerogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, and Candida albicans respectively against concentrates of dimethyl ether, 70% ethanol, and methanol used as controls. The outcome of this investigation demonstrated the significance of enzyme application in drug discovery toward alleviating challenges from pathogens causing progressive diseases that are influencing healthcare and overall environmental declination.

Pectin classifications, mode of action and factors influencing pectinase production

Pectin classifications, mode of action and factors influencing pectinase production

Submerged and solid state fermentation for pectinase production

Submerged and solid state fermentation for pectinase production

Probing the potential of salinity-tolerant endophytic bacteria to improve the growth of mungbean [Vigna radiata (L.) Wilczek

Soil salinity is one of the major limiting factors in plant growth regulation. Salinity-tolerant endophytic bacteria (STEB) can be used to alleviate the negative effects of salinity and promote plant growth. In this study, thirteen endophytic bacteria were isolated from mungbean roots and tested for NaCl salt-tolerance up to 4%. Six bacterial isolates, TMB2, TMB3, TMB5, TMB6, TMB7 and TMB9, demonstrated the ability to tolerate salt. Plant growth-promoting properties such as phosphate solubilization, indole-3-acetic acid (IAA) production, nitrogen fixation, zinc solubilization, biofilm formation and hydrolytic enzyme production were tested in vitro under saline conditions. Eight bacterial isolates indicated phosphate solubilization potential ranging from 5.8-17.7 μg mL-1, wherein TMB6 was found most efficient. Ten bacterial isolates exhibited IAA production ranging from 0.3-2.1 μg mL-1, where TMB7 indicated the highest potential. All the bacterial isolates except TMB13 exhibited nitrogenase activity. Three isolates, TMB6, TMB7 and TMB9, were able to solubilize zinc on tris-minimal media. All isolates were capable of forming biofilm except TMB12 and TMB13. Only TMB2, TMB6 and TMB7 exhibited cellulase activity, while TMB2 and TMB7 exhibited pectinase production. Based on in vitro testing, six efficient STEB were selected and subjected to the further studies. 16S rRNA gene sequencing of efficient STEB revealed the maximum similarity between TMB2 and Rhizobium pusense, TMB3 and Agrobacterium leguminum, TMB5 and Achromobacter denitrificans, TMB6 and Pseudomonas extremorientalis, TMB7 and Bradyrhizobium japonicum and TMB9 and Serratia quinivorans. This is the first international report on the existence of A. leguminum, A. denitrificans, P. extremorientalis and S. quinivorans inside the roots of mungbean. Under controlled-conditions, inoculation of P. extremorientalis TMB6, B. japonicum TMB7 and S. quinivorans TMB9 exhibited maximum potential to increase plant growth parameters; specifically plant dry weight was increased by up to 52%, 61% and 45%, respectively. Inoculation of B. japonicum TMB7 displayed the highest potential to increase plant proline, glycine betaine and total soluble proteins contents by 77%, 78% and 64%, respectively, compared to control under saline conditions. It is suggested that the efficient STEB could be used as biofertilizers for mungbean crop productivity under saline conditions after field-testing.

Pectinase Production by Rhizopus stolonifer A3 Isolated from Apple Peels

The application of pectinase in industries will continue to be developed. To broaden the applicability of enzyme manufacturing in new industries, more research to investigate pectinolytic microorganisms with high activity and stability is still required. Therefore, this study aimed to obtain pectinolytic fungi that have the potential as pectinase producers. Pectinolytic fungi were isolated from rotten apple peels and selected based on clear zone formation on pectic agar media after cetyl trimethyl ammonium bromide (CTAB) staining. Pectinolytic fungi were identified based on 18S rRNA partial gene sequences and morphological characteristics. Pectinase production used Mandels and Weber medium with citrus pectin 20 g l<sup>-1</sup> addition. Pectinase activity was determined based on the measurement of reducing sugars by a colorimetric method. A total of 5 fungal isolates were successfully isolated. All of the isolates had pectinolytic activities with clear zone diameters ranging between 0.99 to 7.32 mm. The isolate A3 showing the highest pectinolytic activity was identified as <em>Rhizopus stolonifer</em>. Microscopically, <em>R</em>. <em>stolonifer </em>A3 showed typical characteristics for <em>Rhizopus</em>, characterized by the presence of rhizoids, stolons and sporangiophores bearing a single spherical sporangium. The pectinase production of <em>R.</em> <em>stolonifer </em>A3 was optimum at initial pH 5.0, temperature 35 °C and incubation period of 3 days with pectinase activity of 14.75 U ml<sup>-1</sup>. <em>R. stolonifer </em>A3 produced acidic pectinase having optimum activity at pH 5.0 and temperature 50 °C. Thus <em>R.</em> <em>stolonifer </em>A3 has the potential to be used as a producer of acidic pectinase appropriate for use in the processing of fruit products.

Cloning and expression of pectinase gene from two forest soil bacteria, Streptomyces sp.

Pectinase is a prominent industrial enzyme with a high market demand and huge applications in diverse avenues. There are different sources for pectinase production; however, a highly stable microbial pectinase that can withstand different physiochemical conditions is needed. Therefore, this study explored the thermostability of the pectinase protein cloned and expressed in Escherichia coli BL21(DE3) from two forest soil bacteria (S-5 and S-14) identified as Streptomyces sp. The pectinase gene illustrated about 750 bp on the gel after electrophoresis and encoded a product of approximately 25 kDa molecular weight. Both expressed pectinase proteins showed optimal activity at 2 different pHs (pH 5 and 9) and 50 ℃. When both recombinant pectinases were exposed to different temperatures, they were able to retain their activity for 120 min indicating both have important thermostable properties. These enzymes could have great potential in industrial biotechnological processes due to their stability over broad pH and high temperatures.

Saccharification of Agricultural Wastes and Clarification of Orange Juice by Penicillium rolfsii CCMB 714 Pectinase

Pectinases are enzymes used in several industrial processes. Seven agroindustrial wastes—jackfruit seed meal (Artocarpus heterophyllus), cocoa seed peel (Theobroma cacao), cocoa husks (Theobroma cacao), passion fruit husks (Passiflora edulis), mangosteen husks (Garcinia mangostana), malt residue (Hordeum vulgare) and the peach palm waste (Bactris gasipaes Kunth.)—were evaluated to produce a crude extract containing pectinase activity by Penicillium rolfsii CCMB 714. The jackfruit seed meal was chosen as the best substrate for solid-state fermentation, which was optimized with 4 mL of water as a wetting agent for 2 days at 35 °C and with a 0.5% nitrogen source, whereby the pectinase production increased by 44% (362.09 U/g). The obtained crude extract was characterized and applied to wastes saccharification and orange juice clarification. The pectinase showed better activity at a pH of 3.0 to 5.0 and 55 °C, it stably maintained over 80% of activity at 30–50 °C for up to 60 min and 1 mM CuSO4 increased the pectinase activity by 17%. The saccharification of agroindustrial wastes (cocoa husks, mangosteen husks and passion fruit husks) resulted in 126.55 µmol/mL of reducing sugars from passion fruit husks, which represents an increase of 126% after optimization (45 °C for 22 h). For the clarification of orange juice, it was possible to reduce the absorbance of the juice by 55%. These results elucidate the potential of the low-cost pectinase solution from P. rolfsii CCMB 714 cultivated in jackfruit seed meal for both the enzymatic pretreatment of plant biomass and the application in beverage industries.