Optimization, purification, and characterization of xylanase production by a newly isolated Trichoderma harzianum strain by a two-step statistical experimental design strategy

Dhaver, Priyashini; Pletschke, Brett I.; Sitholé, Bruce; Govinden, Roshini

doi:10.1038/s41598-022-22723-x

Cited by 27 publications

(12 citation statements)

References 64 publications

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“…The contour plots (Figs. 2 b, 3 b, 4 b) display an elliptical shape indicating that all the parameters interact with each other and are dependent on one another an increase in activity is represented by the peach-shaded regions 15 . Figure 2 a,b illustrates the combined effects of incubation time and glycerol concentration.…”

Section: Resultsmentioning

confidence: 99%

“…Temperature and pH are the most important factors governing microbial growth. Carbon and nitrogen source supplementation provides an enriched environment for microbial growth thus increasing enzyme production 15 . Screening and optimization of growth conditions are crucial to ensure maximal enzyme production with the potential to reduce β -glucosidase production costs 16 .…”

Section: Introductionmentioning

confidence: 99%

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A glucose tolerant β-glucosidase from a newly isolated Neofusicoccum parvum strain F7: production, purification, and characterization

Singh

Sitholé

Kumar

et al. 2023

Sci Rep

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Cellulase-producing microorganisms produce low titres of β-glucosidases with low tolerance to glucose. This study aimed to improve production, purify, and characterize a β-glucosidase from a newly isolated Neofusicoccum parvum strain F7. β-Glucosidase production was significantly enhanced by a sequential statistical modelling approach from 1.5-fold in Plackett–Burman design to 2.5 U/ml in the Box–Behnken design compared to the preliminary one variable at a time experiments (1.6 U/ml). The optimal conditions for enzyme production by BBD were 12 days of fermentation at 20 °C, 175 rpm, 0.5% glycerol and 1.5% casein in pH 6.0 buffer. Three β-glucosidase isoforms referred to as Bgl1, Bgl2, Bgl3 were purified and characterized from the optimized crude extract displaying IC50 values of 2.6, 22.6 and 319.5 mM for glucose, respectively. Bgl3 with a molecular mass of approximately 65 kDa demonstrated the highest tolerance to glucose among the isoforms. The optimum activity and stability for Bgl3 was observed at pH 4.0 in 50 mM sodium acetate buffer with 80% β-glucosidase residual activity retained for three hours. This isoform also retained 60% residual activity at 65 °C for one hour which was then reduced to 40% which remained stable for another 90 min. The β-glucosidase activity of Bgl3 was not enhanced after the addition of metal ions in assay buffers. The Km and vmax for 4-nitrophenyl-β-d-glucopyranoside were 1.18 mM and 28.08 µmol/min, respectively indicating high affinity for the substrate. The ability to withstand the presence of glucose in conjunction with its thermophilic nature indicates promise for this enzyme in industrial application.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A glucose tolerant β-glucosidase from a newly isolated Neofusicoccum parvum strain F7: production, purification, and characterization

Singh

Sitholé

Kumar

et al. 2023

Sci Rep

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“…Because of the heterogeneity and complex chemical nature of plant xylan. Xylan is hydrolyzed by a package of xylanolytic enzymes involving β-1,4-endoxylanase that cooperate for the complete breakdown of xylan into its constituent sugars 17,18 . Xylanase characteristics vary according to their source which makes them useful for an application or another.…”

Section: Introductionmentioning

confidence: 99%

Xylanolytic Biotechnological Applications of Extracellular Xylanase Isolated from Marine Bacillus Subtilis AKM1

Mahmoud

Elhakim

Abdelhamid

et al. 2023

Preprint

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Xylanase enzyme from marine bacteria was investigated for production improvement and studying its effect on agriculture wastes for use in biotechnological applications. By identification of the selective strain by using 16SRNA, the isolate was a new strain called Bacillus Subtilis AKM1 and has the accession number MK377251. The optimum culture conditions for xylanase production were as follows: incubation period (48 hrs.), initial pH (7.0), agitation speed (100 rpm), and incubation temperature(40°C). After purification, xylanase showed maximal activity at pH 7.0 and 50 °C. Metal ions such as Mn2+, Ca2+, and Fe2+ increased the residual activity by increasing the concentration, while Ag+, SDS, and EDTA completely inhibited the activity at higher concentrations. the molecular weight was 34 KDa, Km 0.044 mM, and Vmax 22.222. Xylooligosaccharides produced by enzymatic hydrolysis using the purified xylanase showed antimicrobial activity, antioxidant activity, and anti-tumor activity against breast and colon cancer cells.

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“…Temperature and pH are the most important factors governing microbial growth. Carbon and nitrogen source supplementation provides an enriched environment for microbial growth thus increasing enzyme production [13] . Screening and optimisation of growth conditions are crucial to ensure maximal enzyme production with the potential to reduce β-glucosidase production costs [23] .…”

Section: Introductionmentioning

confidence: 99%

Purification and Characterization of a Glucose tolerant β-Glucosidase from a Newly Isolated Neofusicoccum Parvum strain F7: Production Optimization using Plackett Burman and Box Behnken

Singh

Sitholé

Kumar

et al. 2023

Preprint

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Endoglucanases, exoglucanases and β-glucosidases act synergistically to hydrolyse cellulose into glucose monomers. Thus, this study aimed to improve production of a β-glucosidase from a newly isolated Neofusicoccum parvum strain F7 by optimizing the culture conditions and medium components using Plackett-Burman Design (PBD) and Box Behnken Design (BBD). β-Glucosidase production was significantly enhanced (p-value≤0.05) by 1.5-fold to 2.5 U/ml by BBD as compared to the preliminary one variable at a time (OVAT) experiments of (1.6 U/ml). The optimal conditions for enzyme production by BBD were 12 days of fermentation at 20°C, 175 rpm, 0.5% glycerol and 1.5% casein in 50 mM sodium phosphate (pH 6.0) buffer. Three β-glucosidase isoforms referred to as Bgl1, Bgl2, Bgl3 were purified and characterized from the optimized crude extract displaying IC50 values of 2.6, 22.6 and 319.5 mM for glucose, respectively. Bgl3 with a molecular weight of approximately 65 kDa displayed the highest tolerance to glucose among the isoforms. The optimum activity and stability for Bgl3 was observed at pH 4.0 in 50 mM sodium acetate buffer with 80% β-glucosidase residual activity retained for three hours. This isoform also retained 60% residual activity at 65°C for one hour which was then reduced to 40 % which remained stable for another 90 minutes. The β-Glucosidase activity of Bgl3 was not enhanced after the addition of metal ions in assay buffers. The Km and vmax for 4-nitrophenyl-β-D-glucopyranoside were found to be 1.18 mM and 28.08 µmol/min, respectively indicating high affinity to the substrate. The ability to withstand the presence of glucose in conjunction with its thermophilic nature indicates promise for the enzyme in industrial application.

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Optimization, purification, and characterization of xylanase production by a newly isolated Trichoderma harzianum strain by a two-step statistical experimental design strategy

Cited by 27 publications

References 64 publications

A glucose tolerant β-glucosidase from a newly isolated Neofusicoccum parvum strain F7: production, purification, and characterization

A glucose tolerant β-glucosidase from a newly isolated Neofusicoccum parvum strain F7: production, purification, and characterization

Xylanolytic Biotechnological Applications of Extracellular Xylanase Isolated from Marine Bacillus Subtilis AKM1

Purification and Characterization of a Glucose tolerant β-Glucosidase from a Newly Isolated Neofusicoccum Parvum strain F7: Production Optimization using Plackett Burman and Box Behnken

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