Ficus sycomorus latex: An efficient alternative Egyptian source for horseradish peroxidase in labeling with antibodies for immunodiagnostic kits

Abdel‐Aty, Azza M.; Hamed, Mohamed Belal; Gad, Abdul Aziz M.; El-Hakim, Amr E.; Mohamed, Saleh A.

doi:10.14202/vetworld.2018.1364-1370

Cited by 12 publications

(4 citation statements)

References 27 publications

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“…Precipitation of the proteins in the supernatant was, where applicable, carried out with trichloroacetic acid (TCA) (final concentration of 20% w/v; 4 °C). Proteins were separated by SDS-PAGE and as a standard, the Precision Plus Protein TM Standard (All Blue) from Bio-Rad was used (90, 91). Proteins were visualized by silver staining or by Western blotting and immuno-detection with specific antibodies in combination with a suitable secondary alkaline phosphatase-conjugated antibody (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Enhanced protein secretion in reduced genome strains ofStreptomyces lividans

Hamed

Busche

Simoens

et al. 2023

Preprint

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S. lividans TK24 is a popular host for the production of small molecules and for the secretion of heterologous proteins. TK24 has a large genome with at least 29 secondary metabolite gene clusters that are non-essential for viability and undergo complex regulation. To optimize heterologous protein secretion, we previously constructed ten chassis strains that are devoid of several secondary metabolite gene clusters. Genome reduction was aimed at reducing carbon flow to secondary metabolites and pigmentation in the spent growth medium and improving colony morphology. Strains RG1.0-RG1.10 contain various deletion combinations of the blue actinorhodin cluster (act), the calcium-dependent antibiotic (cda), the undecylprodigiosin (red) and coelimycin A (cpk) clusters, the melanin cluster (mel), the matAB genes that affect mycelial aggregation and the non-essential sigma factor hrdD that controls the transcription of Act and Red regulatory proteins. Two derivative strains, RG1.5 and 1.9, showed a ~15% reduction in growth rate, >2-fold increase in the total mass yield of their native secretome and altered abundance of several specific proteins compared with TK24. Metabolomics and RNAseq analysis revealed that genome reduction led to rapid cessation of growth due to aminoacid depletion and caused both redox and cell envelope stresses, upregulation of the Sec-pathway components secDF and chaperones and a cell envelope two component regulator. RG1.9 maintained elevated heterologous secretion of mRFP and mTNFα by 12-70%. An integrated model is presented linking genome reduction and enhanced secretion.

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Section: Methodsmentioning

confidence: 99%

Enhanced protein secretion in reduced genome strains ofStreptomyces lividans

Hamed

Busche

Simoens

et al. 2023

Preprint

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“…Although the maximum catalytic activity of most of the studied plant POXs were in the range of 40-65 °C, there were several plant POXs which passed this border. For instance, Ficus sycomorus pox was able to retain about 77% of its initial activity at 70 °C after 30 min incubation (Abdel-Aty et al 2018).…”

Section: Physicochemical Properties Of Obpoxmentioning

confidence: 99%

A potent peroxidase from solid cell culture of Ocimum basilicum with high sensitivity for blood glucose determination

Mohammadnejad

Asl

Rasoulian

et al. 2021

Plant Cell Tiss Organ Cult

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Objectives Extensive applications of peroxidase (POX) have raised the global market demand at a considerable rate during the forecast period of 2020 -2025. Nonetheless, the large-scale POX preparation still relies on the extraction from agricultural products, while there is an accumulative driving force toward employing biotechnological processes with agricultural hassle free identity. In pursuit of this trend; Results, A novel heme peroxidase was puri ed to homogeneity (MW of 40 kD) from the callus culture of Ocimum basilicum L. in darkness on Murashige-Skoog medium supplemented by 2,4dichlorophenoxyacetic acid (10 -6 M) and kinetin (10 -5 M). The highest activity of the puri ed peroxidase (ObPOX) was observed in Tris-base buffer at pH 7.5 and 80 °C. ObPOX showed high stability over pH(s) 5 to 7.5 and temperatures of 15 to 60 °C. ObPOX speci c activity was 1245.142 AU mg -1 in the presence of phenol, 4 times higher than that of HRP. ObPOX showed moderate a nity for guaiacol (K m = 21.5 mM), but obtained an exceptionally high speci city constant (k cat /K m = 66743.7 s -1 M -1 ) for GASA (4-[(4-Hydroxy-3-methoxyphenyl) azo]-benzenesulfonic acid), the introduced substrate for determination of blood sugar. Applying ObPOX instead of HRP in glucose measurements of the real samples improved the regression constant of the correlation diagram between the tests and the lab results from 0.958 to 0.981.Conclusion Physicochemical properties of ObPOX as well as the growth rate of basil callus (5.04 g L -1 per day) and the yield of ObPOX production (35 mg per 100 g dry biomass per subculture) designates O. basilicum cell culture for large-scale production of a robust peroxidase.

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“…(Hiraga et al., 2011; Mohamed et al., 2014; Shigeto & Tsutsumi, 2015). They are also essential in medicinal, immunological, biochemical, biotechnological, bioremediation, industrial, and analytical applications (Abdel‐Aty et al., 2018; Mohamed et al., 2017; Pandey et al., 2017).…”

Section: Introductionmentioning

confidence: 99%

“…Among plant peroxidases, horseradish peroxidase (HRP) is commonly used for phenol removal. However, other plant peroxidases have better characters for large scale bioremediation such as low cost purification, low susceptibility to inactivation, broad substrate specificities, wide pH and thermal stability, high yield, and economic values, therefore, they could be alternative choices for horseradish peroxidase (Abdel‐Aty et al., 2013, 2018; González et al., 2008; Mohamed et al, 2011a; Pandey et al., 2017). In addition, the produced peroxidases during seed germination have unique properties to perform a protective role by consuming the accumulated H 2 O 2 and other ROS coupled to oxidation of various phenolic compounds besides their high ability to polymerize various phenolic compounds (Kawano, 2003; Wojtyla et al., 2016; Zhang et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of peroxidases from garden cress sprouts and their roles in lignification and removal of phenol and p ‐chlorophenol

et al. 2020

Self Cite

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The study aims to evaluate the relation between peroxidases of day-6 garden cress sprouts and phenolic compounds. Three cationic, three anionic, and two unbounded peroxidases were separated from day-6 garden cress sprouts. Cationic (GCP1) and anionic (GCP2) peroxidases were purified with molecular masses of 25 and 40 kDa, respectively. The K m values of GCP1 toward H 2 O 2 and guaiacol were lower than GCP2. The anionic GCP2 exhibited high affinity toward some lignin monomers, sinapyl alcohol, coniferyl alcohol, cinnamic and ferulic acids. Therefore, GCP2 is considered as a lignin peroxidase and contributed in lignin synthesis. The activity of GCP1 and GCP2 was stable at a wide pH range 5.5-8.0 and 6.0-7.5, respectively. Both peroxidases showed the same thermal stability range 20-50°C. GCP2 was more resistant against the effect of metal ions than GCP1. GCP2 showed high ability to remove of phenol and p-chlorophenol from effluent compared to GCP1. Practical applications Generally, garden cress is used as a test plant to conduct biomonitoring of pollution in urban soil on a wide scale because of its simplicity, sensitivity, and cost-effectiveness. Peroxidase is an important antioxidant enzyme, which elevated when plant subjected to pollution. Recently, we reported that the increase of peroxidase activity was strongly correlated with high phenolic content and antioxidant activity during the germination of garden cress. In the present study, anionic peroxidase GCP2 may play an important role in lignification process and removal of phenol and p-chlorophenol from polluted soil/wastewater as well as resisted the harmful effect of heavy metals. Cationic peroxidase GCP1, as a natural scavenger, had high affinity toward H 2 O 2 coupled to oxidation of some plant phenolic compounds suggesting its role in consuming of excess H 2 O 2 .

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Ficus sycomorus latex: An efficient alternative Egyptian source for horseradish peroxidase in labeling with antibodies for immunodiagnostic kits

Cited by 12 publications

References 27 publications

Enhanced protein secretion in reduced genome strains ofStreptomyces lividans

Enhanced protein secretion in reduced genome strains ofStreptomyces lividans

A potent peroxidase from solid cell culture of Ocimum basilicum with high sensitivity for blood glucose determination

Purification and characterization of peroxidases from garden cress sprouts and their roles in lignification and removal of phenol and p ‐chlorophenol

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