Physiological roles of histidine acid phytase from Pantoea sp. 3.5.1.

Microbial phytases represent a potential alternative way to produce myo-inositol phosphate isomers with therapeutic properties, whereas the chemical synthesis of these compounds is inefficient and costly. The aim of this work is to study the physiological role of histidine acid phytase in Pantoea sp 3.5.1 cells. Promoter region of the phytase gene was studied and potential binding sites for transcription factors RpoD15 and MetJ were identified by the methods of bioinformatic analysis. Genomic locus carrying the phytase gene was characterised and compared with the same genomic loci of other enterobacteria. It was found that the isolated and purified phytase from Pantoea sp. 3.5.1 refers to the Agp- group of histidine acid phytases and has dual physiological role in the bacterial cell. As glucose-1-phosphatase it is involved in glycolysis processes. The data obtained by stereospecifity analysis helped to identify that phytase from Pantoea sp. 3.5.1 is the enzyme that carries out phytate hydrolysis by the second path and forms D/L-myo-inositol-1,2,4,5,6-pentakisphosphate as the end product

Cloning of Phytase Genes from Pantoea Sp. 3.5.1 and Bacillus ginsengihumi M2.11 in Pichia pastoris

© 2018, Springer Science+Business Media, LLC, part of Springer Nature. Phytases (myo-inositol hexakisphosphate hydrolase) catalyze the hydrolysis of phytate to inorganic phosphate and less phosphorylated myo-inositol derivatives and are widely used as feed additives in animal nutrition. Nevertheless, nowadays, there is a constant search for new phytases and new expression systems for better production of these important enzymes. In this study, we report cloning of two novel bacterial phytases belonging to the different enzyme classes and having different properties in the methylotrophic yeast Pichia pastoris. Sequences of agpP and phyC genes, encoding histidine acid phytase from Pantoea sp. 3.5.1 and β-propeller phytase from Bacillus ginsengihumi M2.11, respectively, were optimized and chemically synthesized according to the P. pastoris codon usage bias. The optimized genes were cloned into the yeast vectors pPINK-HC and pPINK-LC under the control of the inducible promoter AOX1 and two different signal peptides—signal sequence of α-amylase gene from Aspergillus niger and presequence of inulinase gene from Kluyveromyces maxianus. PCR analysis, restriction analysis, and DNA sequencing confirmed correct integration of agpP and phyC genes into P. pastoris genome. As a result, recombinant strains of P. pastoris with codon-optimized bacterial phytase genes (agpP and phyC), encoding histidine acid and β-propeller phytases, integrated into the genome under the alcohol oxidase promoter AOX1 and two different signal peptides, were obtained. Recombinant phytase AgpP was stably expressed and secreted into the culture medium of yeasts, whereas the expression of phyC gene was only confirmed at transcription level

Heterologous expression of Bacillus ginsengihumi phytase gene

Phytate is the most abundant reservoir of phosphorus, which can potentially be used by microorganisms, plants and animals. Microbial phytases, capable of hydrolysing insoluble phytates, can be used as animal feed additives. In the present study, we isolated new phytase-producing bacteria - Bacillus ginsengihumi M2.11, which was identified using Microbial Identification System and 16S rRNA sequencing. We first cloned and sequenced the 1149 bp open reading frame encoding Bacillus ginsengihumi M2.11 phytase into the expression vector pET-46 Ek/LIC by the LIC-cloning method (cloning without ligase). The recombinant E.coli strain expressing Bacillus ginsengihumi M2.11 phytase was obtained. The molecular weight of phytase was 41 kDa

Cloning and Heterologous Expression of Phytase Gene from Pantoea sp. 3.5.1

© 2016, Springer Science+Business Media New York.Phytases (myo-inositol hexakisphosphate phosphohydrolase) catalyzes the stepwise hydrolysis of phosphate groups from phytic acid (myo-inositol hexakisphosphate) or its salt phytate. These enzymes could be potentially used for the stereospecific and efficient production of different myo-inositol phosphate isomers with therapeutic features. In the present study, we cloned the 1728 bp open reading frame encoding Pantoea sp. 3.5.1 phytase into the expression vector pET28a. The recombinant Escherichia coli BL21 pLysS pET28a/agpP strain expressing Pantoea sp. 3.5.1 AgpP phytase was obtained

Digitalization of Energy: from Process Automation to Digital Transformation of the Industry

В данной статье поднимается важный и актуальный на сегодняшний день вопрос - цифровизация энергетики. В данной работе подробно разобраны те отрасли топливо-энергетического комплекса, в которых уже происходит активное внедрение цифровых и интеллектуальных технологий. Перечислен ряд нововведений, который уже используется в энергетике России. Выделены основные недостатки цифровизации.This article raises an important and relevant issue today - the digitalization of energy. In this paper, we analyze in detail those sectors of the fuel and energy complex in which there is already an active introduction of digital and intelligent technologies. A number of innovations that are already being used in the Russian energy sector are listed. The main disadvantages of digitalization are highlighted

The association between <i>Candida albicans</i> sensitivity to antimycotic drugs and the architecture of their microbial community in the oropharynx of HIV infected patients

Relevance. Candida infection remains relevant due to the wide spread of antimycotic-resistant strains of Candida fungi, especially among immunocompromised individuals. It was previously discovered that the Candida spp. microbial community in the biotope of the oropharynx of HIV infected patients is characterized by a certain architecture: they can be present in this biotope in the form of a monoculture or as association of co-isolates. It has been suggested that the architecture of the Candida microbial community may influence their resistance to antimycotic drugs. Purpose a comparative study of the association between the architecture of the C. albicans microbial community in the oropharynx of HIV infected patients with oropharyngeal candidiasis and their sensitivity to antimycotic drugs. Materials and methods. A microbiological study of 52 isolates of Candida fungi (C. albicans, C. glabrata, C. tropicalis and C. krusei) from the oropharynx of 31 HIV infected patients with clinical manifestations of oropharyngeal candidiasis aged 20 to 69 years with almost equal gender distribution was carried out. In the form of monocultures, 18 isolates were isolated, while 34 were identified as co-isolates, which formed 16 homogeneous communities that included strains of the same species, and 18 heterogeneous ones that consisted of fungi of various species. Results. It was found that heterogeneous communities of C. albicans were markedly distinguished by sensitivity to antimycotic drugs, in particular, by low sensitivity to imidazoles. Homogeneous communities practically did not differ from monocultural ones. The general properties of the C. non-albicans population were largely similar to those of C. albicans, but were characterized by lower heterogeneity in response to antimycotic drugs. Conclusion. The architecture of the community of C. albicans isolated from the oropharynx of HIV infected patients with clinical manifestations of oropharyngeal candidiasis affects their sensitivity to antimycotic drugs. When selecting effective antimycotic therapy for such patients, it is necessary to take into account the structure of the Candida spp. community in the oropharynx

Phytate-hydrolyzing rhizobacteria: abiotic stress tolerance and antimicrobial activity

© The Authors 2020. Phytate-hydrolyzing bacteria Pantoea sp. 3.1, 3.2, 3.5.2, 3.6.1 and Bacillus ginsengihumi M2.11 were previously isolated from the soil samples of the Republic of Tatarstan. The effect of cultivation conditions on the growth dynamics as well as antimicrobial activity was determined. All four Pantoea strains showed optimum growth at 26 °C and 28 °C and pH 6.0-7.0. The optimum conditions for the growth of B. ginsengihumi M2.11 strain was determined to be 26 °C, 28 °C and 37 °C and alkaline pH 7 and 8. Salt concentration in the range of 0 to 1000 mM did not significantly affect the growth of the strains. Antagonistic activity of Pantoea sp. 3.5.2 was studied against phytopathogenic micromycetes, identified as Alternaria alternata and Bipolaris sorokiniana. In the presence of bacterial isolate growth of A. alternata was inhibited by 57% and growth of B. sorokiniana - by 85%. Minor growth inhibition by Pantoea sp. 3.5.2 of gram-negative bacteria from Enterobacteriaceae family was observed. The presence of fungicidal activity in the Pantoea strain together with its ability to hydrolyze soil phytates and overcome abiotic stress factors in soil can possibly serve as the basis for the new fungicide of microbial origin

Physiological roles of histidine acid phytase from Pantoea sp. 3.5.1.

Microbial phytases represent a potential alternative way to produce myo-inositol phosphate isomers with therapeutic properties, whereas the chemical synthesis of these compounds is inefficient and costly. The aim of this work is to study the physiological role of histidine acid phytase in Pantoea sp 3.5.1 cells. Promoter region of the phytase gene was studied and potential binding sites for transcription factors RpoD15 and MetJ were identified by the methods of bioinformatic analysis. Genomic locus carrying the phytase gene was characterised and compared with the same genomic loci of other enterobacteria. It was found that the isolated and purified phytase from Pantoea sp. 3.5.1 refers to the Agp- group of histidine acid phytases and has dual physiological role in the bacterial cell. As glucose-1-phosphatase it is involved in glycolysis processes. The data obtained by stereospecifity analysis helped to identify that phytase from Pantoea sp. 3.5.1 is the enzyme that carries out phytate hydrolysis by the second path and forms D/L-myo-inositol-1,2,4,5,6-pentakisphosphate as the end product

Physiological roles of histidine acid phytase from Pantoea sp. 3.5.1.

Microbial phytases represent a potential alternative way to produce myo-inositol phosphate isomers with therapeutic properties, whereas the chemical synthesis of these compounds is inefficient and costly. The aim of this work is to study the physiological role of histidine acid phytase in Pantoea sp 3.5.1 cells. Promoter region of the phytase gene was studied and potential binding sites for transcription factors RpoD15 and MetJ were identified by the methods of bioinformatic analysis. Genomic locus carrying the phytase gene was characterised and compared with the same genomic loci of other enterobacteria. It was found that the isolated and purified phytase from Pantoea sp. 3.5.1 refers to the Agp- group of histidine acid phytases and has dual physiological role in the bacterial cell. As glucose-1-phosphatase it is involved in glycolysis processes. The data obtained by stereospecifity analysis helped to identify that phytase from Pantoea sp. 3.5.1 is the enzyme that carries out phytate hydrolysis by the second path and forms D/L-myo-inositol-1,2,4,5,6-pentakisphosphate as the end product