Purification, biochemical characterization, and genetic cloning of the phytase produced by &lt;i&gt;Burkholderia&lt;/i&gt; sp. strain a13

Graminho, Eduardo Rezende; Takaya, Naoki; Nakamura, Akira; Hoshino, Takayuki

doi:10.2323/jgam.61.15

Cited by 10 publications

(4 citation statements)

References 37 publications

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“…To date, there are limited reports on phytate-specific phytases from Lactobacilli. Broad-specific phosphatases were characterized from Lactobacillus spp., L. sanfranciscensis , and L. plantarum with higher specific activities of 120.2 U/mg and 0.815 U/mg for p -nitrophenyl phosphate as compared to 96.8 U/mg and 0.463 U/mg obtained for sodium phytate (Graminho et al 2015 ; Escobin-Mopera et al 2012 ).…”

Section: Resultsmentioning

confidence: 99%

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Characterization of a putative metal-dependent PTP-like phosphatase from Lactobacillus helveticus 2126

Priyodip

Balaji

2023

Int Microbiol

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To date, there are very limited reports on sequence analysis and structure-based molecular modeling of phosphatases produced by probiotic bacteria. Therefore, a novel protein tyrosine-like phosphatase was characterized from L. helveticus 2126 in this study. The purified bacterial phosphatase was subjected to mass spectrometric analysis, and the identity of constructed sequence was analyzed using peptide mass fingerprint. The 3-D structure of protein was elucidated using homology modeling, while its stability was assessed using Ramachandran plot, VERIFY 3D, and PROCHECK. The bacterium produced an extracellular phosphatase of zone diameter 15 ± 0.8 mm on screening medium within 24 h of incubation. This bacterial phosphatase was highly specific towards sodium phytate as it yielded the lowest Km value of 299.50 ± 4.95 μM compared to other phosphorylated substrates. The activity was effectively stimulated in the presence of zinc, magnesium, and manganese ions thereby showing its PTP-like behavior. The phosphatase showed a molecular mass of 43 kDa, and the corresponding M/Z ratio data yielded 46% query coverage to Bacillus subtilis (3QY7). This showed a 61.1% sequence similarity to Ligilactobacillus ruminis (WP_046923835.1). The final sequence construct based on these bacteria showed a conserved motif “HCHILPGIDD” in their active site. In addition, homology modeling showed a distorted Tim barrel structure with a trinuclear metal center. The final model after energy minimization showed 90.9% of the residues in the favorable region of Ramachandran’s plot. This structural information can be used in genetic engineering for improving the overall stability and catalytic efficiency of probiotic bacterial phosphatases.

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Section: Resultsmentioning

confidence: 99%

“…Phytate was also used as the limiting nutrient for Burkholderia sp. (Graminho et al 2015 ), Klebsiella pneumoniae (Escobin-Mopera et al 2012 ), and Bacillus subtilis (Kerovuo et al 1998 ), showing specific phytase activities of 1.6 U/mg, 5.64 U/mg, and 8 U/mg, respectively.…”

Section: Resultsmentioning

confidence: 99%

Characterization of a putative metal-dependent PTP-like phosphatase from Lactobacillus helveticus 2126

Priyodip

Balaji

2023

Int Microbiol

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“…Many enzymes with phytase activity can be obtained or expressed in plants, animals, and microorganisms [ 18 , 36 , 37 ]. They are classified into the following four groups according to their catalytic mechanisms: histidine acid phosphatases (HAPs), PAPs, Cys or β -helix phosphatases, and protein tyrosine phosphatase-like enzymes (PTP) [ 38 ].…”

Section: Characterization Of Phytasementioning

confidence: 99%

Characterization of the Catalytic Structure of Plant Phytase, Protein Tyrosine Phosphatase-Like Phytase, and Histidine Acid Phytases and Their Biotechnological Applications

et al. 2018

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Phytase plays a prominent role in monogastric animal nutrition due to its ability to improve phytic acid digestion in the gastrointestinal tract, releasing phosphorus and other micronutrients that are important for animal development. Moreover, phytase decreases the amounts of phytic acid and phosphate excreted in feces. Bioinformatics approaches can contribute to the understanding of the catalytic structure of phytase. Analysis of the catalytic structure can reveal enzymatic stability and the polarization and hydrophobicity of amino acids. One important aspect of this type of analysis is the estimation of the number of β-sheets and α-helices in the enzymatic structure. Fermentative processes or genetic engineering methods are employed for phytase production in transgenic plants or microorganisms. To this end, phytase genes are inserted in transgenic crops to improve the bioavailability of phosphorus. This promising technology aims to improve agricultural efficiency and productivity. Thus, the aim of this review is to present the characterization of the catalytic structure of plant and microbial phytases, phytase genes used in transgenic plants and microorganisms, and their biotechnological applications in animal nutrition, which do not impact negatively on environmental degradation.

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“…Phytases -are a class of phosphatases that sequentially hydrolyze phytic acid to less phosphorylated myo-inositol, with the release of inorganic phosphate [2]. Monogastric animals such as pigs, poultry and fish cannot effectively use phytate-bound phosphorus in plant-based feeds due to the limited activity of endogenous intestinal phytates.…”

Section: Introductionmentioning

confidence: 99%

Expression of Bacillus ginsengihumi M2.11 bacterial phytase by recombinant Pichia pastoris strains

2021

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Phytic acid is the main storage form of organic phosphorus. Due to its structural features, phosphorus in phytate is inaccessible for assimilation by animals. Moreover, remaining inaccessible reservoir of phosphorus for animal nutrition, phytic acid is capable of forming insoluble complex salts, which lead to soil and water pollution. Мicrobial enzymes - phytases, capable of decomposing phytic acid to organic phosphorus are being used as feed additives in animal nutrition to solve this problem. Thus, search and development of technologies for the production of enzymes on an industrial scale are the most urgent. Methylotrophic yeast P. pastoris are widely used in biotechnology, as an efficient system for the recombinant proteins expression. They have many advantages, including rapid growth on inexpensive media, a wide range of molecular tools for genetic manipulation in optimizing production processes, they are safe for humans and animals, carry-out many post-translational modifications and produce recombinant proteins intracellularly or extracellularly within a short period of time. It was found that the recombinant P. pastoris strains pPINK-LC-α-MF -phyC, pPINK-HC-α-amyl -phyC, pPINK-LC-α-amyl -phyC, pPINK-HC-α-MF -phyC are able to produce and to secrete B. ginsengihumi bacterial phytase M 2.11 phyC. The maximum activity was observed in the pPINK-LC-α-MF strain – 2.6 (U / mg). Recombinant B. ginsengihumi M 2.11 phytases exhibited high activity in a wide pH range from 2.5 to 9.0. The MF-phyC-HC construction is pH stable. The temperature optimum of all recombinant phytases corresponds to 37 ° C; recombinant phytases retain their activity in the range from -80 to 90C.

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Purification, biochemical characterization, and genetic cloning of the phytase produced by <i>Burkholderia</i> sp. strain a13

Cited by 10 publications

References 37 publications

Characterization of a putative metal-dependent PTP-like phosphatase from Lactobacillus helveticus 2126

Characterization of a putative metal-dependent PTP-like phosphatase from Lactobacillus helveticus 2126

Characterization of the Catalytic Structure of Plant Phytase, Protein Tyrosine Phosphatase-Like Phytase, and Histidine Acid Phytases and Their Biotechnological Applications

Expression of Bacillus ginsengihumi M2.11 bacterial phytase by recombinant Pichia pastoris strains

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