Development of reproducible assays for polygalacturonase and pectinase

Li, Qian; Coffman, Anthony M.; Ju, Lu‐Kwang

doi:10.1016/j.enzmictec.2015.02.006

Cited by 33 publications

(14 citation statements)

References 36 publications

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“…The enzymatic activity of the pectinase cocktail was determined by the rate of release of reducing ends from polygalacturonate, according to methods described previously (Li et al, 2015; Reid and Ricard, 2000) with some modifications Briefly, two test tubes containing 2 mL of diluted enzyme stock were equilibrated to 55°C and 10 mL of substrate stock solution was added to each tube. Tubes with or without HHP treatment (50 MPa) were incubated in a water bath at 55°C for 0.5, 1, 2, 3, or 4 h. Solution A was prepared by sequentially dissolving Na 2 CO 3 , glycine, and CuSO 4 in distilled water and was used as a time 0 control.…”

Section: Methodsmentioning

confidence: 99%

Utilization of Pectinase Cocktail and High Hydrostatic Pressure for the Production of Aged Black Garlic Juice with Improved Nutritional Value

Kim¹,

Kim

et al. 2019

pnf

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In comparison with raw garlic, aged black garlic has been shown to display multiple pharmacological activities. We recently reported that pretreatment of pectinase cocktail with high hydrostatic pressure (HHP) before the process of aging garlic juice improves its antidiabetic activity and increases S-allylcysteine (SAC) content. Thus, this study was designed to investigate the influence of pectinase cocktail with HHP on the quality of aged black garlic juice formation and to identify the optimal manufacturing conditions. In the pretreatment step, garlic juice is heated at 55°C for 24 h. The contents of SAC and total polyphenols were increased with treatment of pectinase cocktail; this increase was greater under HHP processing. In contrast, the total flavonoid content was decreased in all pretreatment conditions. Garlic juice pretreated with pectinase cocktail and HHP had a significantly higher content of SAC in the early phase of aging than raw garlic juice, and the SAC was increased over time in both treatment groups. The total polyphenol content of garlic juice was significantly higher in the pretreatment group during the aging period, and the antioxidant activity of garlic juice showed a positive correlation with polyphenol content. Interestingly, HHP increased the enzymatic activity of the pectinase cocktail.

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Section: Methodsmentioning

confidence: 99%

Utilization of Pectinase Cocktail and High Hydrostatic Pressure for the Production of Aged Black Garlic Juice with Improved Nutritional Value

Kim¹,

Kim

et al. 2019

pnf

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“…and 4 °C) to obtain the supernatant for further study. Polygalacturonase/ pectinase activity was calculated by measuring the concentration of reducing sugar using the modified dinitrosalicylic acid(DNS) method using pectin as the substrate 28 . Pectinase acts on pectin and releases galacturonic acid, which reduces DNS to amino nitrosalicylic acid with lmax of 540 nm.…”

Section: Pectinase Assay and Protein Estimationmentioning

confidence: 99%

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

Meena¹,

Sowmeya²,

Praveen³

et al. 2021

J Pure Appl Microbiol

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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.

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“…The procedure for xylanase estimation is similar to cellulase with the substrate being 0.06% oat spelt xylan. Polygalacturonase assay reaction mixture comprised polygalacturonic acid 0.45%, culture supernatant 0.1 ml in 50 mM acetate buffer pH 5 [11].…”

Section: Fungal Strain and Culture Conditionsmentioning

confidence: 99%

Transcriptomic analysis ofLentinus squarrosulusprovide insights into its biodegradation ability

Ravichandran

Kolte

Dhali

et al. 2020

Preprint

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Basidiomycetes are of special interest in biotechnological research for their versatile potential in degradation of lignocellulosic biomass. This study accordingly reports analysis of transcriptome of a white-rot Basidiomycete L.squarrosulus grown in simple potato dextrose broth supplemented with aromatic compound, reactive black dye to gain an insight into the degradation ability of the fungus. RNA was sequenced using Illumina NextSeq 500 to obtain 6,679,162 high quality paired end reads that were assembled de novo using CLC assembly cell to generate 25,244 contigs.Putative functions were assigned for the 10,494 transcripts based on sequence similarities through BLAST2GO 5.2 and Function annotator. Functional assignments revealed enhanced oxidoreductase activity through the expression of diverse biomass degrading enzymes and their corresponding co-regulators. CAZyme analysis through dbCAN and CUPP revealed the presence of 6 families of polysaccharide lyases, 51 families of glycoside hydrolases, 23 families of glycoside transferases, 7 families of carbohydrate esterases and 10 families of Auxiliary activities.Genes encoding the ligninolytic enzymes and auxiliary activities among the transcript sequences were identified through gene prediction by AUGUSTUS and FGENESH. Biochemical analysis of a couple of biomass degrading enzymes substantiated the functional predictions. In essence, L.squarrosulus grown in a simple medium devoid of lignocellulosic substrate demonstrated presence of a repertoire of lignocellulose degrading enzymesimplying that source of lignocellulose is not required for expression of these biomass degrading enzymes. The study hereby underlines the significance of L.squarrosulus in biomass degradation and its future functional exploitation in biomass conversion applications.

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Development of reproducible assays for polygalacturonase and pectinase

Cited by 33 publications

References 36 publications

Utilization of Pectinase Cocktail and High Hydrostatic Pressure for the Production of Aged Black Garlic Juice with Improved Nutritional Value

Utilization of Pectinase Cocktail and High Hydrostatic Pressure for the Production of Aged Black Garlic Juice with Improved Nutritional Value

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

Transcriptomic analysis ofLentinus squarrosulusprovide insights into its biodegradation ability

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