Concentrations Of Medium Components Research Articles

Investigation of Crypthecodinium cohnii High-Cell-Density Fed-Batch Cultivations

Crypthecodinium cohnii is a marine microalga that can accumulate high amounts of polyunsaturated fatty acids (PUFAs) and thus replace conventional routes of fish oil production. They are associated with the destruction of marine resources and multiple downstream/purification complications. The major drawbacks of using C. cohnii for industrial-scale production are associated with low PUFA productivity. One of the means of increasing the PUFA synthesis rate is to maintain the medium component concentrations at optimal values throughout cultivation, thus increasing PUFA production efficiency, which can result in the successful transfer of the process to pilot and/or industrial scale. The goal of the present research was to develop techniques for increasing the efficiency of PUFA production via C. cohnii cultivation. Multiple experiments were carried out to test and fine-tune the cultivation medium composition and oxygen transfer factors. The biomass yields from individual components, yeast extract, sea salts, and glucose amounted to 5.5, 0.65, and 0.61 g·g−1, respectively. C. cohnii cell susceptibility to mechanical damage was experimentally evaluated. Power inputs of <276.5 W/m3 did not seem to promote cell destruction when Pitched-blade impellers were used. The obtained cultivation conditions were shown to be efficient in terms of increasing the biomass productivity and the omega-3 fatty acid content in C. cohnii. By using the applied methods, the maximal biomass productivity reached 8.0 g·L−1·day−1, while the highest obtained biomass concentration reached 110 g·L−1. A steady increase in the concentration of PUFAs during cultivation was observed from the FTIR data.

Production of fungal phytases in solid state fermentation and potential biotechnological applications.

Phytases are important enzymes used for eliminating the anti-nutritional properties of phytic acid in food and feed ingredients. Phytic acid is major form of organic phosphorus stored during seed setting. Monogastric animals cannot utilize this phytate-phosphorus due to lack of necessary enzymes. Therefore, phytic acid excretion is responsible for mineral deficiency and phosphorus pollution. Phytases have been reported from diverse microorganisms, however, fungal phytases are preferred due to their unique properties. Aspergillus species are the predominant producers of phytases and have been explored widely as compared to other fungi. Solid-state fermentation has been studied as an economical process for the production of phytases to utilize various agro-industrial residues. Mixed substrate fermentation has also been reported for the production of phytases. Physical and chemical parameters including pH, temperature, and concentrations of media components have significantly affected the production of phytases in solid state fermentation. Fungi produced high levels of phytases in solid state fermentation utilizing economical substrates. Optimization of culture conditions using different approaches has significantly improved the production of phytases. Fungal phytases are histidine acid phosphatases exhibiting broad substrate specificity, are relatively thermostable and protease-resistant. These phytases have been found effective in dephytinization of food and feed samples with concomitant liberation of minerals, sugars and soluble proteins. Additionally, they have improved the growth of plants by increasing the availability of phosphorus and other minerals. Furthermore, phytases from fungi have played an important roles in bread making, semi-synthesis of peroxidase, biofuel production, production ofmyo-inositol phosphates and management of environmental pollution. This review article describes the production of fungal phytases in solid state fermentation and their biotechnological applications.

Simultaneous Production of Cellulase and β-Carotene in the Filamentous Fungus Trichoderma reesei.

β-Carotene is an indispensable additive in beverage, cosmetic, feed, and pharmaceutical production. The fermentation industry annually generates abundant waste mycelia from Trichoderma reesei (T. reesei), a pivotal industrial strain for cellulase and heterologous protein production. In this study, we constructed a T. reesei cell factory for β-carotene production for the first time. Four key enzymes, CarRP, CarB, GGS1/CrtE, and HMG1, were overexpressed in T. reesei. The concentrations of medium components, including tryptone and glucose, were optimized. The modified strain accumulated β-carotene at a titer of 218.8 mg/L in flask culture. We achieved cellulase production (FPase, 22.33 IU/mL) with the concomitant production of β-carotene (286.63 mg/L) from T. reesei in a jar. Overall, this study offers a novel and unique approach to address the costly waste mycelium management process using T. reesei industrial strains that simultaneously produce proteins and carotenoids.

Optimization of Xanthan Gum Production by Demerara Sugar Using Response Surface Methodology

Xanthan gum (XG) production using three Xanthomonas sp. strains (290, 472, and S6) was evaluated by applying a 23 full factorial central composite design response to study the interactive effects of the fermentation medium component concentrations as parameters to determine the efficiency of the gum production in batch experiments. The experimental variables were the carbon source (demerara sugar or sucrose), potassium phosphate dibasic, and magnesium sulfate. Experimental results showed the K2HPO4 concentration as the important parameter for XG production by using Xanthomonas axonopodis pv. manihotis IBSBF 290 and X. campestris pv. campestris IBSBF 472, while for the Xanthomonas sp. S6 strain, the MgSO4∙7H2O concentration was the determining factor in XG production using demerara sugar or sucrose as a carbon source. The strains of Xanthomonas 472 and S6, using demerara sugar and higher concentrations of salts, exhibited a higher yield of XG (36 and 32%) than when using sucrose and the same concentration of salts. The experimental outcomes highlighted demerara sugar as a suitable and efficient alternative carbon and micronutrient source for XG production. Despite the bacterial strain influence, the medium composition is crucial for this fermentation process. Therefore, the evaluated salts are important factors for XG production, and the demerara sugar can partially replace this mineral salt requirement as indicated by the face-centered composite experimental design due to its chemical composition. Overall, demerara sugar provides promising properties for XG production.

БИОСТОЙКОСТЬ ЦЕМЕНТНЫХ КОМПОЗИТОВ ИЗ СУХИХ СТРОИТЕЛЬНЫХ СМЕСЕЙ

A study was made of the influence of a model biological environment of the products of the metabolism of filamentous fungi on the biostability of cement composites from industrial dry building mixes for outdoor and indoor work. The low resistance of samples without additional bioprotection to the influence of the biological environment was established. Relevant is the development of cement composites with biocidal additives to ensure resistance to biologically and chemically aggressive environments without reducing the strength properties. The degree of fouling of samples from various types of dry building mixtures in a model environment of metabolic products of filamentous fungi was determined. Eight composites of different composition were studied under the influence of 13 types of model medium. The stability of the samples was established at a concentration of medium components from 0 to 5% by weight. Tile adhesive "UNIS 2000", tile adhesive "Prospectors", putty "Prospectors", facade plaster "KNAUF" have fungus-resistant properties. Putty "CT29 Ceresit". Waterproofing "VodoStopGlims", plaster "IvsilGross" and self-leveling floor "Magma" are non-mushroom resistant. The most unfavorable concentrations of filamentous fungi metabolism products for the samples were revealed. The effectiveness of the use of polymer-based biocidal additives of the Teflex series (OOO SoftPROTECTOR, Russia) on the resistance of cement composites to biodegradation was revealed. The additives "Anti-Salt Wash", "For Metal" and "Universal" showed the greatest efficiency, their use provides fungi resistance and fungicidal properties of building materials.

APPLICATION OF RESPONSE SURFACE METHODOLOGY (RSM) TO ENHANCE MCLPHA COPOLYMER PRODUCTION BY MASSILIA SP USING CANE MOLASSES AS A CARBON SOURCE

Cane molasses is an inexpensive, sustainable by-product of the sugar industry, which supports the growth of microorganisms as it contains several growth factors. Response Surface Methodology (RSM) employing central composite design (CCD) was applied to enhance PHA production by Massilia sp at shake flask level by using molasses as a carbon source. Isolate produced maximum DCW (54.0 g/l) and PHA (3.0 g/l) when medium was optimized with the help of CCD. Optimized concentration of medium components to maximize PHA production was found to be as follows: molasses: 100.0 g/l, NH4HPO4 : 1.67g/l, Na2HPO4 : 5.86g/l, K2HPO4 : 5.74g/l, MgSO4 : 0.25g/l, trace element solution: 0.1ml/l. The point optimization method reported a small difference of 0.02g/l, indicating the validity of mathematical models developed after applying CCD.

Impact of preparation pH and temperature on amino acid stability of highly concentrated cell culture feed media

AbstractBACKGROUNDThe stability of amino acids (AAs) in a highly concentrated cell culture medium is affected by medium preparation and storage conditions. Variability in the concentration of medium components (including AAs) can influence cell culture performance and product quality attributes.RESULTSIn this report, the impact of preparation pH, preparation temperature, and storage temperature was evaluated on the stability of 16 AAs in a highly concentrated cell culture feed medium. Optimizing the medium pH enabled an increase in the initial feed medium concentration by 30% plus 3.9 g L−1 tyrosine with no observed precipitation after 28 days. Up to 6.0 g L−1 tyrosine can be supplemented in the medium at ~pH 10 without precipitation. Moreover, preparing the medium at 40–50 °C prevented precipitation during storage at 4–8 °C. A slight negative impact on AA stability during this high‐temperature preparation was observed, but this impact was negligible relative to the AA degradation observed during extended storage of media prepared at 37 °C. The optimal preparation pH and temperature allowed the concentrated medium to be stored at 4–8 °C with minimal AA degradation and without precipitation. These discoveries were further supported by principal component analysis of AA profile similarity before and after storage at different pH and temperature conditions.CONCLUSIONThis study describes the medium preparation parameters to improve AA stability and mitigate precipitation issues. The optimization strategy will facilitate the development of robust cell culture production processes. © 2022 Society of Chemical Industry (SCI).

Polyaniline-Pectin nanoparticles immobilized paper based colorimetric sensor for detection of Escherichia coli in milk and milk products

In the food quality and safety arena, there is a need to develop novel and sustainable methodologies that can help in the prevention of foodborne diseases. Herein, we report the development of a rapid conducting polymer strip-based sensor using Polyaniline-pectin (PANI-PEC) for the detection of Escherichia coli in milk and milk products. Polyaniline-pectin nanoparticles stabilized with biopolymer pectin were synthesized and its characterization studies such as FTIR, UV-Vis spectroscopy, electrical conductivity and particle size analysis were done. The assay parameters were optimized for the selective detection of E. coli in milk and milk products. The concentration of PANI-PEC solution immobilized/strip was optimized to be 3 mg/mL as it exhibited good sensitivity and colour intensity. Based on acid production and selectivity for E.coli, concentrations of media components like lactose, tryptophan, yeast extract, chondroitin sulphate, sodium lauryl sulphate, potassium chloride, tergitol-7, gentamycin sulphate and ampicillin trihydrate were optimized as 0.9, 0.1, 0.45, 0.015, 0.1, 2, 0.0125, 0.00016 and 0.015 respectively and sample volume was optimized to 500 μL. The developed PANI-PEC colorimetric strip-based sensor detects 0.52 ± 0.17 log CFU/mL E. coli within 10: 21 h (h). Further shelf-life study revealed that the developed PANI-PEC colorimetric sensor strips are stable at room temperature up to six months exhibiting the same sensitivity. The results obtained here indicate that this novel and simple paper based colorimetric sensor holds potential for application in food industries as a reliable and rapid method for detection of E. coli in milk and milk products at various stages of production and processing.

Statistical optimization of medium components for biosurfactant production by Pseudomonas guguanensis D30

Biosurfactant production by Pseudomonas guguanensis D30 was reported using mineral oil in submerged condition. Twelve medium components were tested at two levels by Plackett–Burman design, among them, mineral oil, yeast extract, peptone, MgSO4, and CaCl2 found significant on the basis of emulsification index. These five significant components were further optimized through central composite design (CCD). The experimental design was successfully used for regression analysis and the significant model suggested the solution of 10% (v/v) mineral oil, 3.0 g/L (w/v) yeast extract and 0.2 g/L (w/v) peptone for 13.14 g/L predicted biosurfactant production. We kept the suggested concentrations of medium components and got 13.34 ± 0.08 g/L biosurfactant production, which is almost double the conventional one-factor-at-a-time production (7.126 ± 0.12 g/L). It reduced the surface tension of the medium up to 28 ± 1.2 mN/m. We found ethyl acetate a suitable solvent for biosurfactant extraction amongst methanol, chloroform, and methanol:chloroform. The partially purified biosurfactant was chemically characterized as lipopeptide by Fourier transform infrared spectroscopy (FT-IR).

Simulated annealing and artificial neural network as optimization tools to enhance yields of bioemulsifier and exopolysaccharides by thermophilic Brevibacillus borstelensis

Bioemulsifiers and exopolysaccharides are industrially important biomolecules that are produced by microbial fermentation and have applications in food, cosmetics and pharmaceutical industries. Both the compounds are produced extracellularly as secondary metabolites by microorganisms. In the current work, two statistical optimization tools, simulated annealing and artificial neural network, were used for optimized production of bioemulsifier and exopolysaccharide by thermophilic Brevibacillus borstelensis. The process variables were concentrations of glucose, monosodium glutamate, yeast extract and magnesium sulfate in production medium and concentrations of bioemulsifier, exopolysaccharide and biomass were studied as responses. The best function values for the three responses as obtained by simulated annealing were 7.14, 17.87 and 3.3 while the model generated by ANN predicted 6.1 g/l, 17.5 g/l and 2.5 g/l as the values of the three responses. The study shows the efficiency of statistical optimization tools compared to the experimental methods for optimization of bioprocess to maximize yield. These techniques also reduce the time and cost of any bioprocess. There is a rise in agricultural residues and the problems associated with them. Agricultural wastes contain high amounts of carbon and nitrogen and hence can be used as suitable substrates for the production of bioemulsifier and exopolysaccharide. The concentrations of medium components obtained from optimized media will be useful when agricultural wastes are used as substrates. This will not only lead to production of two industrially important compounds from sustainable sources but will also result in environmental remediation.

Development of a cost-effective medium for submerged production of fungal aryl alcohol oxidase using a genetically modified Aspergillus nidulans strain

Aryl alcohol oxidase (AAO), an extracellular H2O2-providing enzyme, plays a central role in lignin depolymerization. Cost-effective production of AAO has not been achieved, due to the low yield of enzyme-producing microorganisms and the high cost of fermentation media. This study aims to develop a cost-effective medium for high-yield production of AAO in submerged culture using a recombinant Aspergillus nidulans strain. Results demonstrate that corn steep liquor (CSL) was a rich but inexpensive nitrogen source for AAO production, and CSL can provide enough trace metals and vitamins (i.e. pyridoxine) for A. nidulans. A 2-level Plackett-Burman design was utilized to determine the main affecting factors in AAO production. The medium was further optimized by a 3-level Box-Behnken design to obtain the optimum medium component concentrations (61.0 g/L maltose, 26.4 g/L CSL, and 13.8 g/L NaNO3). The greatest AAO activity achieved was 1021 U/L with a protein concentration of 0.75 g/L.

Improvement of delta-endotoxin production from local Bacillus thuringiensis Se13 using Taguchi’s orthogonal array methodology

Abstract Objective The insecticidal activity of Bacillus thuringiensis directly depends on the yield of delta-endotoxins. In this study, various nutritional and cultural parameters influencing delta-endotoxin synthesis by a local isolate of B. thuringiensis Se13 were investigated using Taguchi methods. Methods In the first experiment, four factors, incubation period, incubation temperature, initial pH and medium, each at four levels, were selected and an orthogonal array layout of L16 was carried out. In the second experiment, Taguchi’s orthogonal array method of L27 was used to evaluate the effects of the different concentration of medium components. Taguchi’s signal–noise ratio and variance analysis were applied to determine the effect of the factors. After each experiment, verification studies were carried out using determined optimum conditions. Results The optimum conditions for incubation period, incubation temperature, initial pH, and medium determined as 72 h, 30°C, pH 9, and M4 medium, respectively. In the second experiment, soybean flour (5%), glucose (5%), KH2PO4 (0.3%), K2HPO4 (0.1%), MgSO4 (0.4%) were determined as the optimum conditions. The delta-endotoxin yield was elevated to 1559.25 μg mL−1 when the factors were adjusted to optimum level. Conclusion Optimization using the Taguchi method appeared to be a good choice for the overproduction of delta-endotoxin.

Optimization of Nutrient Composition for Producing ACE Inhibitory Peptides from Goat Milk Fermented by Lactobacillus bulgaricus LB6.

Hypertension is a serious threat to human health and food-derived angiotensin converting enzyme (ACE; EC 3.4.15.1) inhibitory peptides can be used to regulate high blood pressure without side effects. The composition of the nutrient medium for the production of these peptides by fermenting goat milk with Lactobacillus bulgaricus LB6 was optimized to increase the ACE inhibitory activity by Box-Behnken design (BBD) of response surface methodology (RSM) in the present study. Soybean peptone, glucose, and casein had significant effects on both ACE inhibition rate and viable counts of L. bulgaricus LB6 during incubation. The results showed that the maximum values of ACE inhibition rate and viable counts for L. bulgaricus LB6 were reaching to 86.37 ± 0.53% and 8.06 × 107 under the optimal conditions, which were 0.35% (w/w) soybean peptone, 1.2% (w/w) glucose, and 0.15% (w/w) casein. The results were in close agreement with the model prediction. The optimal values of the medium component concentrations can be a good reference for obtaining ACE inhibitory peptides from goat milk.

Dextran Utilization During Its Synthesis by Weissella cibaria RBA12 Can Be Overcome by Fed-Batch Fermentation in a Bioreactor.

Weissella cibaria RBA12 produced a maximum of 9mg/ml dextran (with 90% efficiency) using shake flask culture under the optimized concentration of medium components viz. 2% (w/v) of each sucrose, yeast extract, and K2HPO4 after incubation at optimized conditions of 20°C and 180rpm for 24h. The optimized medium and conditions were used for scale-up of dextran production from Weissella cibaria RBA12 in 2.5-l working volume under batch fermentation in a bioreactor that yielded a maximum of 9.3mg/ml dextran (with 93% efficiency) at 14h. After 14h, dextran produced was utilized by the bacterium till 18h in its stationary phase under sucrose depleted conditions. Dextran utilization was further studied by fed-batch fermentation using sucrose feed. Dextran on production under fed-batch fermentation in bioreactor gave 35.8mg/ml after 32h. In fed-batch mode, there was no decrease in dextran concentration as observed in the batch mode. This showed that the utilization of dextran by Weissella cibaria RBA12 is initiated when there is sucrose depletion and therefore the presence of sucrose can possibly overcome the dextran hydrolysis. This is the first report of utilization of dextran, post-sucrose depletion by Weissella sp. studied in bioreactor.

Optimization of Culture Medium for Maximal Production of Spinosad Using an Artificial Neural Network - Genetic Algorithm Modeling

Background: Spinosyns, products of secondary metabolic pathway of Saccharopolyspora spinosa, show high insecticidal activity, but difficulty in enhancing the spinosad yield affects wide application. The fermentation process is a key factor in this case. Methods: The response surface methodology (RMS) and artificial neural network (ANN) modeling were applied to optimize medium components for spinosad production using S. spinosa strain CGMCC4.1365. Experiments were performed using a rotatable central composite design, and the data obtained were used to construct an ANN model and an RSM model. Using a genetic algorithm (GA), the input space of the ANN model was optimized to obtain optimal values of medium component concentrations. Results: The regression coefficients (R²) for the ANN and RSM models were 0.9866 and 0.9458, respectively, indicating that the fitness of the ANN model was higher. The maximal spinosad yield (401.26 mg/l) was obtained using ANN/GA-optimized concentrations. Conclusion: The hybrid ANN/GA approach provides a viable alternative to the conventional RSM approach for the modeling and optimization of fermentation processes.

OPTIMIZATION OF CULTURAL MEDIA FOR JOINT CULTIVATION OF BRADYRHIZOBIUM JAPONICUM AND AZOSPIRILLUM BRASILENSE

The paper presents the results of studies on the optimization of culture media for joint cultivation of Bradyrhizobium japonicum and Azospirillum brasilense bacteria at periodic passages. It was established that cultivation of studied microorganisms in a mixed culture the number of nodule bacteria vary from 0.76 to 6.72 billion cells/cm3, depending on the concentration of medium components. It was shown that the effect of molasses, fodder yeast and phosphates on growth activity of B. japonicum significantly depends not only on the amount of chemical components in the media, but also on the strain of A. brasilense, used for joint cultivation. Based on the results of multivariate experiment an optimized media for joint cultivation of studied microorganisms was proposed.

OPTIMIZATION OF MEDIUM COMPOSITION FOR cis,cis-MUCONIC ACID PRODUCTION BY A Pseudomonas sp. MUTANT USING STATISTICAL METHODS

cis,cis-Muconic acid (CCMA) is used as a platform chemical for the production of several high-value compounds. For this article, an optimization strategy has been used to optimize medium composition for CCMA production from fairly cheap benzoate by Pseudomonas sp. 1167. The effect of different concentrations of medium components on CCMA production was studied. CCMA yields obtained from Plackett–Burman design (PBD) showed wide variation (3.95–5.87 g/L), and the first-order model indicated that (NH4)2SO4 (P < 0.01) and K2HPO4 · 3H2O (P < 0.02) were the significant components for CCMA production. Then the optimization was performed by steepest ascent design (SAD) and central composite design (CCD), and a validation experiment was conducted to verify the predicted value. The optimal medium composition was: 12 g/L sodium benzoate, 2.5 g/L sodium succinate, 0.7932 g/L (NH4)2SO4, 1.5612 g/L K2HPO4 · 3H2O, 1.2 g/L MgSO4 · 7H2O, 0.4 g/L yeast extract, 0.08 g/L FeCl3 · 6H2O, and 0.08 g/L ethylenediamine tetraacetic acid (EDTA). Under these conditions, a maximum of 7.18 g/L CCMA was produced per 12 g/L benzoate with a highly efficient process within 11 hr and a molecular conversion yield of 61%. Altogether, our results provide valuable insights into nutritional supplementation of CCMA production by using statistical methods, which may benefit a cost-competitive industrial fed-batch fermentation process using a cheap substrate.

Process development of eicosapentaenoic acid production

Eicosapentaenoic acid (EPA), a well-known member of omega-3 fatty acids, is considered to have a significant health promoting role in the human body. It is an essential fatty acid as the human body lacks the ability to produce it in vivo and must be supplemented through diet. Microbial EPA represents a potential commercial source. GC/MS analyses confirmed that bacterial isolate 717, similar to Shewanella pacifica on the basis of 16S rRNA sequencing, is a potential high EPA producer. Two types of bioreactors, a Stirred Tank Reactor (STR) and an Oscillatory Baffled Reactor (OBR), were investigated in order to choose the optimum system for EPA production. The EPA production media was optimised through the selection of media components in a Plackett–Burman (PB) design of experiment followed by a Central Composite Design (CCD) to optimise the concentration of medium components identified as significant in the Plackett–Burman experiment. The growth conditions for the bioreactor, using artificial sea water (ASW) medium, were optimised by applying Response Surface Methodology (RSM). This optimisation strategy resulted in an increase in EPA from 33mg/l (10mg/g biomass), representing 8% of the total fatty acids at shake flask level, to 350mg/l (46mg/g biomass) representing 25% of the total fatty acids at bioreactor level. During this study the main effects and the interactions between the bioreactor growth conditions were revealed and a polynomial model of EPA production was generated. Chemostat experiments were performed to test the effect of growth rate and temperature on EPA production.

Metal-tolerant PAH-degrading bacteria: development of suitable test medium and effect of cadmium and its availability on PAH biodegradation

The use of metal-tolerant polyaromatic hydrocarbon (PAH)-degrading bacteria is viable for mitigating metal inhibition of organic compound biodegradation in the remediation of mixed contaminated sites. Many microbial growth media used for toxicity testing contain high concentrations of metal-binding components such as phosphates that can reduce solution-phase metal concentrations thereby underestimate the real toxicity. In this study, we isolated two PAHs-degrading bacterial consortia from long-term mixed contaminated soils. We have developed a new mineral medium by optimising the concentrations of medium components to allow the bacterial growth and at the same time maintain high bioavailable metal (Cd(2+) as a model metal) in the medium. This medium has more than 60 % Cd as Cd(2+) at pH 6.5 as measured by an ion selective electrode and visual MINTEQ model. The Cd-tolerant patterns of the consortia were tested and minimum inhibitory concentration (MIC) derived. The consortium-5 had the highest MIC of 5 mg l(-1) Cd followed by consortium-9. Both cultures were able to completely metabolise 200 mg l(-1) phenanthrene in less than 4 days in the presence of 5 mg l(-1) Cd. The isolated metal-tolerant PAH-degrading bacterial cultures have great potential for bioremediation of mixed contaminated soils.

Optimization of Polygalacturonase Production from a Newly IsolatedThalassospira frigidphilosprofundusto Use in Pectin Hydrolysis: Statistical Approach

The present study deals with the production of cold active polygalacturonase (PGase) by submerged fermentation using Thalassospira frigidphilosprofundus, a novel species isolated from deep waters of Bay of Bengal. Nonlinear models were applied to optimize the medium components for enhanced production of PGase. Taguchi orthogonal array design was adopted to evaluate the factors influencing the yield of PGase, followed by the central composite design (CCD) of response surface methodology (RSM) to identify the optimum concentrations of the key factors responsible for PGase production. Data obtained from the above mentioned statistical experimental design was used for final optimization study by linking the artificial neural network and genetic algorithm (ANN-GA). Using ANN-GA hybrid model, the maximum PGase activity (32.54 U/mL) was achieved at the optimized concentrations of medium components. In a comparison between the optimal output of RSM and ANN-GA hybrid, the latter favored the production of PGase. In addition, the study also focused on the determination of factors responsible for pectin hydrolysis by crude pectinase extracted from T. frigidphilosprofundus through the central composite design. Results indicated 80% degradation of pectin in banana fiber at 20°C in 120 min, suggesting the scope of cold active PGase usage in the treatment of raw banana fibers.