Pectinase Production from Banana Peel Biomass via the Optimization of the Solid-state Fermentation Conditions of Aspergillus niger Strain

In this study, the biomass of banana peel was used to produce pectinase via optimization of solid-state fermentation conditions of the filamentous fungi Aspergillus nigeA. niger). The operating conditions of solid-state fermentation were optimized using the method of full factorial design with incubation temperature ranging between 25 °C and 35 °C, moisture content between 40% and 60%, and inoculum size between 1.6 x 106 spores/mL and 1.4 x 107 spores/mL. Optimizing the solid-state fermentation conditions appeared crucial to minimize the sample used in this experimental design and determine the significant correlation between the operating conditions. A relatively high maximal pectinase production of 27 UmL-1 was attained at 35° C of incubation, 60% of moisture content, and 1.6 x 106 spores/mL of inoculum size with a relatively low amount of substrate (5 g). Given that the production of pectinase with other substrates (e.g., pineapple waste, lemon peel, cassava waste, and wheat bran) generally ranges between 3 U/mL and 16 U/mL (Abdullah et al., 2018; Handa et al., 2016; Melnichuk et al., 2020; Thangaratham and Manimegalai, 2014; Salim et al., 2017), thus the yield of pectinase derived from the banana peel in this study (27 U/mL) was considered moderately high. The findings of this study indicated that the biomass of banana peel would be a potential substrate for pectinase production via the solid-state fermentation of A. niger.

Utilização do bagaço de malte da indústria cervejeira como substrato para produção de pectinase por cogumelos / Utilization of beer spent grain from the brewing industry as substrate for mushroom pectinase production

O aumento da produção de cerveja promoveu, consequente, aumento na geração de resíduos, como o bagaço de malte, provenientes dessa atividade industrial, intensificando a necessidade de seu reaproveitamento. Uma alternativa para isso, é a utilização dos resíduos da indústria cervejeira como fonte de carbono no cultivo de macrofungos (cogumelos), visando a produção de biomoléculas, como enzimas, de importante interesse pelas indústrias. O objetivo desse estudo foi avaliar a produção de pectinase por duas espécies de basiomicetos, Pleurotus djamor e Hypsizygus ulmarius, a partir de cultivo submerso, tendo o bagaço de malte como fonte de carbono e aplicando um delineamento composto central rotacional para otimização das variáveis do processo: concentração de substrato, temperatura e agitação. Para a produção de pectinase por P. djamor, as variáveis que melhor influenciaram o processo foram concentração de bagaço de malte (30 g/L), temperatura (29°C) e agitação (75 rpm), quando obteve-se atividade enzimática de 2,544 U/mL. Já o cultivo de H. ulmarius para produção de pectinase, as variáveis que melhor influenciaram o processo foram concentração do bagaço de malte (30 g/L), temperatura (24°C) e agitação (150 rpm), quando foi verificada atividade enzimática de 2,367 U/mL. Ensaios de estabilidade térmica de pectinase obtidas a partir do cultivo submerso em bagaço de mate, de ambas espécies de basiodiomicetos, indicaram bom desempenho da enzima sob temperaturas mais altas (80°C). Estes resultados, sugerem portanto que o bagaço de mate é uma fonte de carbono interessante a ser aplicada no cultivo submerso de P. djamor e H. ulmarius visando a produção de pectinase .

Isolation and Screening of Pectinase Producing Bacteria from Soil Sample

The vast majority of the industrial use of enzymes is covered from microorganisms. Microorganisms are favoured in industry because of their several advantages for example rapid growth, short life expectancy and simplicity in doing genetic alterations. Microbial enzymes are thus amply provided, very much standardized and promoted by many companies. Among various enzymes, Pectinases hold an exceptional place because of its different uses in various sectors like food, textile and biofuel industries.A total of 25% of total enzyme market is being shared by Pectinase alone.The current study was carried out to evaluate the pectinase activity of the pectinolytic bacteria. 40 Bacterial strains were isolated from different soil samples and screened for Pectinase production. Primary and Secondary screening showed 3 potential isolates I38 , I39 and I40 showing pectin degradation on Vincent’s media. Further, extracellular pectinase was partially purified by ammonium sulphate precipitation and dialysis. Sequential ammonium sulphate saturations from 20-80% i.e. (20, 40, 60 and 80%) showed 60% ammonium sulphate was optimum for precipitation of intracellular enzyme whereas 80% was optimum for extracellular enzyme.

Production and Optimization of Pectinase from Pectinolytic Fungi Cultivated on Mango peels and Pectin Subjected to Submerged Fermentation

Pectinases are the group of enzymes that degrade pectin. This study was conducted with the aim of isolation of efficient pectinase producing pectinolytic fungi from the decomposing mango peels using extracted mango peels pectin as a growth substrate under submerged fermentation, determining optimum pectinase production conditions with regards to some physicochemical parameters. The organisms were screened for the production of pectinase using Pectin agar media, and the two active pectinolytic fungi (P1 and P2) were isolated. pectinase production media was later used for the Lab scale production of pectinase by inoculating p1 and p2 and incubating for 7 days. The enzyme was extracted after seven days of fermentation and every day tested for their pectinolytic activity. P2 showed relatively higher pectinolytic activity and was therefore used for further studies. P2 was inoculated into a broth containing mango pectin under submerged fermentation. Results indicate that a pectin yield of mango peel 17.75%. Different parameters optimization processes were investigated on submerged fermentation namely pH, incubation period, temperature and substrate concentration optima were found 6, 4 days, 35oC and 1.5% respectively. The result suggests that mango peels have high pectin content and can be used for the value-added synthesis of pectinase.

Pectin Degradation in Fruit Juices by Pectinase from Meyerozyma sp. VITPCT75 Isolated from Phyllanthus emblica

This study aimed to identify and characterize a pectinase-producing novel yeast from the fermented juice of Phyllanthus emblica and apply the enzyme for fruit juice clarification. Among the five pectinase-producing yeasts, isolate-1 exhibited the highest pectinase activity and was further used in this study. Based on morphological, physiological, and 18SrRNAanalyses, isolate-1 was recognized as a new strain sharing 99% sequence homology with other Meyerozyma strains and was thus designated as Meyerozyma sp. VITPCT75. The strain produced pectinase optimally at a temperature and pH of 25oC and 7, respectively. Maximum pectinase production was observed after 4-days incubation. The enzyme exhibited optimum activity at the temperature of 25 °C and pH 7.0. The enzyme was more stable at a temperature and pH of 20 °C and 7, respectively. Storage stability studies revealed that the enzyme was stable at -20 °C. The cell-free supernatant was partially purified using ammonium sulfate and solvent precipitation. Acetone at a concentration of 20% assured an adequate partial purification. The molecular weight of pectinase was determined as 6 kDa. The enzymatic metal ion preference-related studies revealed that Ca²z, Kz, Cu²z, Fe²z, and Ba²z ions enhanced, Ni²z ions moderately inhibited, and Mn²z ions intensely inhibited the enzymatic activity. Neither Na+ and Mg2+ ions nor EDTA affected the enzyme activity. When subjected to fruit juice clarification, the enzyme significantly reduced the viscosity of the juice.

Optimization of Pectinase Activity From Locally Isolated Fungi and Agrowastes

Background: Pectin enzymes are biocatalysts that degrade pectin into simpler forms. Fermentation is the commonly utilized method for pectinase production. Prior to optimization of pectinase activity, preliminary findings were undertaken to select the best screened microbe (Aspergillus niger), agrowastes and extraction solvents. Solid-state fermentation was employed in the study (optimization process), utilizing the Box-Behnken design in Design-Expert software package version 12.0.3. Results: The results showed 0.1 molar sodium chloride as the best extraction solvent, with the activity higher than the citrate buffer. However, pectinase activity obtained with distilled water was significantly (p<0.05) lower than 0.1 molar sodium chloride. For the substrates employed in the study, the citrus (orange) peel had the highest pectinase activity of ≈0.40 mg per ml. The activity of citrus peels was significantly higher than the activities from each of corn cob, banana peel, wheat bran, Thaumatococcus danielli fruit wastes and Thaumatococcus danielli leaves. A significant increase (p<0.05) in pectinase activity was also obtained with Thaumatococcus danielli fruit wastes relative to Thaumatococcus danielli leaves. Pichia kudriavzevii strain F2-T429-5 and Pichia kudriavzevii strain CY902 have been identified to complement for pectinase production. From the results obtained, the optimum conditions for pectinase production were approximately 5.87 days of fermentation; pH of 3.90; at 21.24 oC; particle size of 0.06-inch; inoculum volume of 1.00 ml, and agitation time (for pectinase extraction) at 11.43 minutes. Optimisation of the selected conditions using the Box-Behnken design and analysis as a statistical tool resulted in a higher activity. Conclusion: Local production of pectinase would help in reducing hunger through local job creation, thereby positively contributing to the Sustainable Development Goals (SDGs).