Purification, characterization, and cloning of a novel phytase with low pH optimum and strong proteolysis resistance from Aspergillus ficuum NTG-23

Zhang, G.Q.; Dong, Xiaofang; Wang, Z.H.; Zhang, Q.; Wang, H.X.; Tong, Jun

doi:10.1016/j.biortech.2010.01.001

Cited by 75 publications

(69 citation statements)

References 40 publications

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“…Phytase activity was measured using a modified ferrous sulfate-molybdenum blue assay [2]. In brief, 25 μ L enzyme solution was incubated with 475 μ L of 5 mM sodium phytate in 50 mM Tris-HCl buffer (pH 7.0) at 37°C for 15 min.…”

Section: Methodsmentioning

confidence: 99%

“…Phytic acid ( myo -inositol 1, 2, 3, 4, 5, 6-hexakisphosphoric acid) is the primary storage form of phosphate in plants and the prime concern for human nutrition and health management [1, 2]. As an antinutritional component, phytic acid can strongly chelate with cations such as calcium, kalium, magnesium, iron, copper, zinc, and as well as proteins [3].…”

Section: Introductionmentioning

confidence: 99%

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A Phytase Characterized by Relatively High pH Tolerance and Thermostability from the Shiitake MushroomLentinus edodes

Zhang

et al. 2013

BioMed Research International

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A monomeric phytase with a molecular mass of 14 kDa was acquired from fresh fruiting bodies of the shiitake mushroom Lentinus edodes. The isolation procedure involved chromatography on DEAE-cellulose, CM-cellulose, Q-Sepharose, Affi-gel blue gel, and a final fast protein liquid chromatography-gel filtration on Superdex 75. The purified phytase demonstrated the unique N-terminal amino acid sequence DPKRTDQVN, which exhibited no sequence similarity with those of other phytases previously reported. It expressed its maximal activity at pH 5.0 and 37°C. Phytase activity manifested less than 20% change in activity over the pH range of 3.0–9.0, considerable thermostability with more than 60% residual activity at 70°C, and about 40% residual activity at 95°C. It displayed a wide substrate specificity on a variety of phosphorylated compounds with the following ranking: ATP > fructose-6-phosphate > AMP > glucose-6-phosphate > ADP > sodium phytate > β-glycerophosphate. The phytase activity was moderately stimulated by Ca2+, but inhibited by Al3+, Mn2+, Zn2+, and Cu2+ at a tested concentration of 5 mM.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Phytase Characterized by Relatively High pH Tolerance and Thermostability from the Shiitake MushroomLentinus edodes

Zhang

et al. 2013

BioMed Research International

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“…In PhypA, such Ca 2ϩ -dependent activation was not produced. Chelating agents such as EDTA generally have a positive effect ranging from a 10 to a 50% increase in activity (18,19). This was not the case for bifidobacterial phytases, where a slight negative effect of EDTA on activity was monitored (Table 1).…”

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confidence: 99%

Novel Phytases from Bifidobacterium pseudocatenulatum ATCC 27919 and Bifidobacterium longum subsp. infantis ATCC 15697

Tamayo-Ramos

Sanz‐Penella

Yebra

et al. 2012

Appl Environ Microbiol

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Two novel phytases have been characterized from Bifidobacterium pseudocatenulatum and Bifidobacterium longum subsp. infantis. The enzymes belong to a new subclass within the histidine acid phytases, are highly specific for the hydrolysis of phytate, and render myo-inositol triphosphate as the final hydrolysis product. They represent the first phytases characterized from this group of probiotic microorganisms, opening the possibilities for their use in the processing of high-phytate-content foods. Phytases are a special class of phosphatases that catalyze the sequential hydrolysis of myo-inositol hexakis phosphate (InsP 6 ) or phytate, the major form of phosphorous storage in plants, to lower myo-inositol phosphate derivatives. Based on biochemical properties and amino acid sequence, phytases can be categorized into two major classes, the histidine acid phytases and the alkaline phytases (9). The use of commercial phytases of fungal origin (usually Trichoderma and Aspergillus strains) is a common practice in animal feeding for improving the bioaccessibility of minerals (zinc, iron, calcium, and magnesium) that are otherwise chelated by InsP 6 and InsP 5 and insolubilized (4, 13). The presence of phytase activity has recently been reported to occur in the human intestinal Bifidobacterium pseudocatenulatum ATCC 27919 and Bifidobacterium longum subsp. infantis ATCC 15697 strains (5, 7). These bacteria were included in the production process of whole-wheat bread, showing their potential to reduce phytate content in fiber-rich products for human consumption (15, 16). However, no genetic information about the corresponding enzymes was available.Identification and purification of bifidobacterial phytases. When the genomes from B. pseudocatenulatum ATCC 27919 (GenBank accession no. NZ_ABXX02000003) and B. longum subsp. infantis ATCC 15697 (GenBank accession no. CP001095) were being searched for the presence of genes encoding putative phosphatases, two genes (BIFPSEUDO_03792 and BLON_0263, designated hereinafter PhypA and PhylA, respectively) attracted our attention. These genes encoded two proteins with 59% amino acid identity which possessed histidine acid phosphatase (HAP) domains (PF00328). Furthermore, PhypA and PhylA displayed N-terminal signal peptides for secretion and a predicted C-terminal sortase-dependent cell wall-anchoring domain. Initial characterization of the phytase activity in both bifidobacterial strains revealed that it was mainly located at the cell wall/membrane fractions (data not shown). Hence, the identified genes were good candidates for encoding the bifidobacterial phytases.Sequence analysis additionally showed that the typical HAP signature sequence of the histidine-containing catalytic site (RH GXRXP) was replaced by RHGSRGL in PhypA and PhylA and that the aspartate of the C-terminal HD domain was replaced by alanine. Remarkably, the putative HAP homologues closer to PhylA and PhypA found in databases, belonging to the Actinobacteria, Betaproteobacteria, and Gammaproteobacteria, show the same...

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“…In blood pH, it causes mineral deficiencies of metabolism by leading to insoluble mineral-phytate complex formation of zinc, iron, calcium, magnesium, manganese and copper metal ions which both act as enzyme cofactors and as different metabolic processes. Also, phytates completely prevent the formation of the phytate-protein complex of protein and amino acids (Kumar, Sinha, Makkar, & Becker, 2010;Zhang et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Immobilization of a thermostable phytase from Pinar melkior (Lactarius piperatus) onto magnetite chitosan nanoparticles

Önem

Çiçek

Nadaroğlu

2015

CyTA - Journal of Food

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Phytase enzyme was purified from the Pinar melkior (Lactarius piperatus) mushroom using ammonium sulfate precipitation and DEAEsephadex ion exchange chromatography techniques. First, the purified phytase enzyme was covalently bound to the surface of magnetite-CTS NPs in yield of 87%. Then, optimum pHs of the free and immobilized enzymes were determined as 5.0 and 4.0, respectively. Optimum temperature of free and immobilized enzymes was found as 60°C. Also, the effects of some metal ions on activity of free and immobilized phytase enzymes were investigated. Also, research was undertaken as to whether the purified free and immobilized phytase enzyme could hydrolyze the phytic acid in many cereal products or not. And, it was discovered that the immobilized phytase enzyme hydrolyzed the phytic acid at the highest rate (75.02% rate) in wheat. From the findings obtained, that immobilized enzyme was quite resistant to temperature, pH and metal ions.Se purificó la enzima fitasa de la seta Pinar melkior (Lactarius piperatus) utilizando precipitación de sulfato de amonio y técnicas cromatográficas de intercambio iónico DEAE con sephadex. Primeramente, la enzima fitasa purificada fue ligada de forma covalente a la superficie de CTS NP con magnetita con una eficiencia del 87%. Después, el pH óptimo de las enzimas libres e inmovilizadas fue determinado como 5,0 y 4,0 respectivamente. La temperatura óptima de las enzimas libres e inmovilizadas se encontró a 60°C. Además, se investigaron los efectos de algunos iones metálicos en la actividad de las enzimas fitasas libres e inmovilizadas. También, se estudió si las enzimas fitasas libres e inmovilizadas purificadas podrían hidrolizar el ácido fítico en varios productos que contengan cereales. Además, se determinó que las enzimas fitasa inmovilizadas, hidrolizaron el ácido fítico con el valor más alto (75,02%) en el trigo. A partir de estos resultados se encontró que las enzimas inmovilizadas eran muy resistentes a la temperatura, pH e iones metálicos.

show abstract

Purification, characterization, and cloning of a novel phytase with low pH optimum and strong proteolysis resistance from Aspergillus ficuum NTG-23

Cited by 75 publications

References 40 publications

A Phytase Characterized by Relatively High pH Tolerance and Thermostability from the Shiitake MushroomLentinus edodes

A Phytase Characterized by Relatively High pH Tolerance and Thermostability from the Shiitake MushroomLentinus edodes

Novel Phytases from Bifidobacterium pseudocatenulatum ATCC 27919 and Bifidobacterium longum subsp. infantis ATCC 15697

Immobilization of a thermostable phytase from Pinar melkior (Lactarius piperatus) onto magnetite chitosan nanoparticles

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