Purification and characterization of polygalacturonase from Aspergillus fumigatus MTCC 2584 and elucidating its application in retting of Crotalaria juncea fiber

Anand, Gautam; Yadav, Sangeeta; Yadav, Dinesh

doi:10.1007/s13205-016-0517-4

Cited by 35 publications

(21 citation statements)

References 38 publications

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“…However, pH stability range of the enzyme was wider (3.0-11.0) as compared to fungal PGs reported from A. niger (Kant et al 2013). An endo-PG with stability in alkaline range 7.0-11.0 has recently been reported from A. fumigatus (Anand et al 2016). …”

Section: Effect Of Ph On the Activity And Stability Of Pgmentioning

confidence: 98%

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Production, purification and biochemical characterization of an exo-polygalacturonase from Aspergillus niger MTCC 478 suitable for clarification of orange juice

Anand

Yadav

2017

3 Biotech

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Polygalacturonases (PG) represent an important member of pectinases group of enzymes with immense industrial applications. A fungal strain Aspergillus niger MTCC478 was used for the production of polygalacturonase both under submerged and solid-state fermentation condition. Further its production was optimized under solid-state fermentation condition with media comprising of wheat bran and tea extract. Purification of an exo-PG was achieved by acetone precipitation (60-90%) and CMcellulose column chromatography revealing 15.28-fold purification with a specific activity of 33.47 U/mg protein and 1.2% yield. A relative molecular mass of purified PG was approximately 124.0 kDa. The pH and temperature optimum was found to be 4 and 50°C, respectively. The k cat and K m value for degradation of PGA by the purified enzyme was found to be 194 s -1 and 2.3 mg/mL, respectively. Cu 2? was found to enhance the PG activity while Ag ? completely inhibited the enzyme activity. The application of the purified PG in orange juice clarification was elucidated.

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Section: Effect Of Ph On the Activity And Stability Of Pgmentioning

confidence: 98%

“…It has been reported that activity of an acidic endo-PG from A. niger and alkaline endo-PG from A. fumigatus is enhanced by Cu 2? at 1 mM concentration (Zhou et al 2015;Anand et al 2016). Metal ions like Mn 2?…”

Section: Effect Of Metal Ions and Protein Inhibitorsmentioning

confidence: 99%

Production, purification and biochemical characterization of an exo-polygalacturonase from Aspergillus niger MTCC 478 suitable for clarification of orange juice

Anand

Yadav

2017

3 Biotech

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“…It appears that many endo-polygalacturonases belonged to acidic ones with optimal pH values around 3.5-6.0, including endo-TePG28b (pH 3.5) [23], endo-PGA1 (pH 3.5) [21], endo-PG I (pH 3.5) [18], Nfpg II (pH 4.0) [20], PGA-ZJ5A from Aspergillus niger ZJ5 (pH 4.5) [12], PG7fn (pH 5.0) [24], AaEPG (pH 5.0) [15], Nfpg I (pH 5.0) [20], endo-PG I (pH 6.0) [22], and PG2 (pH 6.0) [19]. Only several endo-polygalacturonases were classified as alkaline, such as Endo-PG from Aspergillus fumigatus MTCC 2584 with optimal pH 10.0 [35]. As shown in Figure 2, AnEPG showed the highest activity at pH 4.0, and no obvious activities were detected above pH 6.0.…”

Section: Determination Of Ph Optima and Ph Stability Of Anepgmentioning

confidence: 99%

“…Only several endo-polygalacturonases were classified as alkaline, such as Endo-PG from Aspergillus fumigatus MTCC 2584 with optimal pH 10.0 [35]. The pH stability of AnEPG was also investigated ( Figure 3).…”

Section: Determination Of Ph Optima and Ph Stability Of Anepgmentioning

confidence: 99%

Overexpression and Biochemical Characterization of an Endo-α-1,4-polygalacturonase from Aspergillus nidulans in Pichia pastoris

Zhang

et al. 2020

IJMS

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Pectinases have many applications in the industry of food, paper, and textiles, therefore finding novel polygalacturonases is required. Multiple sequence alignment and phylogenetic analysis of AnEPG (an endo-α-1,4-polygalacturonase from Aspergillus nidulans) and other GH 28 endo-polygalacturonases suggested that AnEPG is different from others. AnEPG overexpressed in Pichia pastoris was characterized. AnEPG showed the highest activity at pH 4.0, and exhibited moderate activity over a narrow pH range (pH 2.0-5.0) and superior stability in a wide pH range (pH 2.0-12.0). It displayed the highest activity at 60 • C, and retained >42.2% of maximum activity between 20 and 80 • C. It was stable below 40 • C and lost activity very quickly above 50 • C. Its apparent kinetic parameters against PGA (polygalacturonic acid) were determined, with the K m and k cat values of 8.3 mg/mL and 5640 µmol/min/mg, respectively. Ba 2+ and Ni 2+ enhanced activity by 12.2% and 9.4%, respectively, while Ca 2+ , Cu 2+ , and Mn 2+ inhibited activity by 14.8%, 12.8%, and 10.2% separately. Analysis of hydrolysis products by AnEPG proved that AnEPG belongs to an endo-polygalacturonase. Modelled structure of AnEPG by I-TASSER showed structural characteristics of endo-polygalacturonases. This pectinase has great potential to be used in food industry and as feed additives.In nature, pectin is degraded by pectin-degrading enzymes [5][6][7][8][9]. Due to complexity and heterogeneousity, complete degradation of pectin requires the combined action of esterases, lyases, and hydrolases [5][6][7][8][9]. Esterases mainly include pectin methylesterases (EC 3.1.1.11), acetylesterases (EC 3.1.1.6), and feruloyl esterases (EC 3.1.1.73), which remove methyl, acetyl, and feruloyl of pectin, respectively. Pectate lyases (EC 4.2.2.2) and pectin lyases (EC 4.2.2.10) catalyze the cleavage of the α-1,4 glycosidic bond of pectin and pectate by transeliminative reactions, respectively, to produce ∆4,5 unsaturated products. Hydrolases cleave the α-1,4-linkage of pectin and pectate, including endo-polygalacturonases (EC 3.2.1.15), exo-polygalacturonases (EC 3.2.1.67), rhamnogalacturonases (EC 3.2.1.171), etc. [5-9]. Among the pectinolytic enzymes, endo-polygalacturonases (endo-PGs) are most extensively investigated, and are classified into family GH 28 of CAZy (carbohydrate-active enzymes database) based on sequence and structure similarity [5-9]. Most characterized endo-PGs are from fungi, such as the species of Aspergillus and Penicillium. For example, some endo-PGs were purified and characterized from different genera of Aspergillus, including A. niger [10-12], A. awamori [13], A. carbonarius [14], A. aculeatus [15], and A. flavus [16]. Some endo-PGs from different species of Penicillium were also investigated, for example, Penicillium oxalicum CZ1028 [17], Penicillium sp. CGMCC 1669 [18], and Penicillium occitanis [19]. Many endo-PGs from other fungi species were characterized as well, for example, Neosartorya fischeri P1 [20], Bispora sp. MEY-1 [21], Achaetomium sp....

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“…It has been reported in the literature that Ca 2+ inhibits the catalytic action of polygalacturonase (Anand, Yadav, & Yadav, ). The present results also showed that Ca 2+ inhibited the catalytic ability of PgaB.…”

Section: Resultsmentioning

confidence: 99%

Identification and characterization of thermostable endo‐polygalacturonase II B from Aspergillus luchuensis

et al. 2020

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Endo‐polygalacturonase II B (PgaB) from Aspergillus luchuensis was orthologous to endo‐polygalacturonase from Aspergillus niger with mutant sites Thr42Ser and Glu52Ala. Mature pgaB gene was cloned from the genomic DNA of A. luchuensis and secreted expressed with over 90% purity in Pichia Pastoris and reached 1.0 g/L after 144 hr culture. The recombinant PgaB was further purified by Ni‐NTA chromatography. Using polygalacturonic acid (PGA) as substrate, the optimal condition for PgaB activity was 40°C and pH 4.5, respectively. Km and Vmax of PgaB were 0.19 mmol/l and 103.58 μmol min−1 mg−1, respectively. The relative activity of PgaB remained more than 60% and 40% of maximum activity at 50 and 60°C for 7 hr. PgaB increased the light transmittance by 85% and showed high efficiency in juice clarification. The main product was galacturonic acid oligosaccharides with degrees of polymers (DP) 1–3. The PgaB is a potential pectinolytic enzyme in food industries. Practical applications Endo‐polygalacturonase II B (PgaB) was identified from Aspergillus luchuensis, a filamentous fungus widely used in food and beverage fermentation in East Asia. PgaB still kept its most activity at 60°C for 7 hr. Polygalacturonic acid (PGA) can be digested effectively by the PgaB and the main products are galacturonic acid oligosaccharides with degrees of polymers (DP) 1–3. PgaB shows high efficiency in juice clarification. The PgaB is a potential pectinolytic enzyme for the applications in food industries.

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Purification and characterization of polygalacturonase from Aspergillus fumigatus MTCC 2584 and elucidating its application in retting of Crotalaria juncea fiber

Cited by 35 publications

References 38 publications

Production, purification and biochemical characterization of an exo-polygalacturonase from Aspergillus niger MTCC 478 suitable for clarification of orange juice

Production, purification and biochemical characterization of an exo-polygalacturonase from Aspergillus niger MTCC 478 suitable for clarification of orange juice

Overexpression and Biochemical Characterization of an Endo-α-1,4-polygalacturonase from Aspergillus nidulans in Pichia pastoris

Identification and characterization of thermostable endo‐polygalacturonase II B from Aspergillus luchuensis

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