非生物因素对枯草芽孢杆菌IMV B-7023植酸酶活性的影响

Q4 Biochemistry, Genetics and Molecular Biology Mikrobiolohichnyi zhurnal Pub Date : 2023-02-28 DOI:10.15407/microbiolj84.06.003
N. Chuiko, A. Chobotarov, I. Kurdish
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The influence of abiotic factors on Bacillus subtilis IMV B-7023 phytase activity has not previously been studied, so this was the aim of our research. Methods. The phytase activity of bacteria was studied by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction, and the nanomaterials’ influence on growth — by cultivation methods. Results. The highest B. subtilis IMV B-7023 phytase activity was observed at 28°C. Also, there was no B. subtilis IMV B-7023 phytase activity at pH 4—6. However, this activity increased at pH 7 and did not change significantly with increasing the buffer system pH to 12. Silicon dioxide influence on the B. subtilis IMV B-7023 growth activity during cultivation in a media with phytate as a phosphorus source depended on the nanomaterial concentration. Thus, at 0.05 and 0.5 g/L of silicon dioxide in the medium, this strain growth activity increased by 8—18%, and at 5.0 g/L of these nanoparticles, bacteria growth inhibition by 19% was observed. At the same time, clay mineral bentonite did not affect the B. subtilis IMV B-7023 growth under the studied cultivation conditions. In addition, silicon dioxide and bentonite stimulated B. subtilis IMV B-7023 phytase activity at all studied concentrations. So, phytase activity increased by 1.82—3.34 times upon adding silicon dioxide and by 2.54—5.83 times upon adding bentonite into the medium. Since the optimal values for phytase activity of most genus Bacillus bacteria are within neutral pH values and temperatures within 50—55°C, a property of B. subtilis IMV B-7023 to show maximum phytase activity at alkaline pH and lower temperatures (28°C) and also stimulation of this activity by soil minerals increases competitiveness of this strain as a component of a bacterial preparation for crop production. Conclusions. Abiotic environmental factors influence the B. subtilis IMV B-7023 supernagrowth and phytase activity. Optimal physicochemical factors for the phytase activity of these bacteria are temperature 28°C and pH 7—12. The concentrations 0.05, 0.5, and 5.0 g/L of silicon dioxide and bentonite increase B. subtilis IMV B-7023 phytase activity. The effect of these nanoscale minerals on the B. subtilis IMV B-7023 growth depends on their type and concentration during cultivation in the medium with phytate as a phosphorus source. 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引用次数: 0

摘要

芽孢杆菌属的细菌可以合成特定的植酸酶。这一特性对土壤细菌尤为重要。它有助于矿化植酸和植酸盐,并为这些细菌和植物(在它们生活的根区)提供可用的磷。我们之前的研究表明枯草芽孢杆菌IMV B-7023菌株具有植酸酶活性,可以利用植酸盐作为营养来源。它是用于作物生产的“偶氮”复合细菌制剂的一个组成部分。众所周知,非生物环境因素会影响细菌的植酸酶活性。特别是在不同的pH和温度下,植酸酶活性变化显著。固体土壤颗粒,包括纳米级矿物质,也会影响细菌的酶活性。非生物因子对枯草芽孢杆菌IMV B-7023植酸酶活性的影响此前未见研究,因此这是我们的研究目的。方法。通过测定植酸钠在酶促反应中释放的磷酸盐量,以及纳米材料对细菌生长的影响,研究了细菌的植酸酶活性。结果。28℃时,枯草芽孢杆菌IMV B-7023植酸酶活性最高。pH值4 ~ 6时,枯草芽孢杆菌IMV B-7023植酸酶无活性。然而,当pH为7时,这种活性增加,当缓冲系统pH增加到12时,这种活性没有显著变化。在植酸为磷源的培养基中,二氧化硅对枯草芽孢杆菌IMV B-7023生长活性的影响取决于纳米材料浓度。因此,在培养基中添加0.05和0.5 g/L二氧化硅时,该菌株的生长活性提高了8-18%,在5.0 g/L二氧化硅纳米颗粒时,细菌的生长抑制率为19%。同时,黏土矿物膨润土对枯草芽孢杆菌IMV B-7023的生长没有影响。此外,二氧化硅和膨润土在所有研究浓度下都能刺激枯草芽孢杆菌IMV B-7023植酸酶活性。因此,添加二氧化硅后植酸酶活性提高了1.82 ~ 3.34倍,添加膨润土后植酸酶活性提高了2.54 ~ 5.83倍。由于大多数芽孢杆菌属细菌的植酸酶活性的最佳值是在中性pH值和50-55°C的温度范围内,枯草芽孢杆菌IMV B-7023在碱性pH值和较低温度(28°C)下显示出最大的植酸酶活性,并且土壤矿物质对这种活性的刺激增加了该菌株作为作物生产细菌制剂的组成部分的竞争力。结论。非生物环境因素对枯草芽孢杆菌IMV B-7023超生长和植酸酶活性的影响。温度28℃、pH 7 ~ 12是这些细菌培养植酸酶活性的最佳理化条件。0.05、0.5和5.0 g/L浓度的二氧化硅和膨润土均能提高枯草芽孢杆菌IMV B-7023植酸酶活性。这些纳米级矿物质对枯草芽孢杆菌IMV B-7023生长的影响取决于它们在以植酸盐为磷源的培养基中培养时的类型和浓度。结果表明,枯草芽孢杆菌IMV B-7023菌株具有在中性和碱性土壤中有效吸收植酸盐的潜在能力,特别是由于这些细菌与膨润土和二氧化硅纳米颗粒的相互作用。这扩大了在农业技术中使用枯草芽孢杆菌IMV B-7023的可能性。
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Abiotic Factors Influence on Bacillus subtilis IMV B-7023 Phytase Activity
Bacteria of the Bacillus genus can synthesize specific phytase enzymes. This property is especially important for soil bacteria. It helps to mineralize phytin and phytates and to provide these bacteria and plants (in the root zone of which they live) with the available phosphorus. Our previous studies have demonstrated that the Bacillus subtilis IMV B-7023 strain exhibits a phytase activity and can use phytate as a nutrition source. It is a component of the «Azogran» complex bacterial preparation for crop production. As known, abiotic environmental factors can influence the phytase activity of bacteria. In particular, the phytase activity changes significantly under different pH and temperatures. Solid soil particles, including nanosized minerals, can also influence bacteria’s enzymatic activity. The influence of abiotic factors on Bacillus subtilis IMV B-7023 phytase activity has not previously been studied, so this was the aim of our research. Methods. The phytase activity of bacteria was studied by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction, and the nanomaterials’ influence on growth — by cultivation methods. Results. The highest B. subtilis IMV B-7023 phytase activity was observed at 28°C. Also, there was no B. subtilis IMV B-7023 phytase activity at pH 4—6. However, this activity increased at pH 7 and did not change significantly with increasing the buffer system pH to 12. Silicon dioxide influence on the B. subtilis IMV B-7023 growth activity during cultivation in a media with phytate as a phosphorus source depended on the nanomaterial concentration. Thus, at 0.05 and 0.5 g/L of silicon dioxide in the medium, this strain growth activity increased by 8—18%, and at 5.0 g/L of these nanoparticles, bacteria growth inhibition by 19% was observed. At the same time, clay mineral bentonite did not affect the B. subtilis IMV B-7023 growth under the studied cultivation conditions. In addition, silicon dioxide and bentonite stimulated B. subtilis IMV B-7023 phytase activity at all studied concentrations. So, phytase activity increased by 1.82—3.34 times upon adding silicon dioxide and by 2.54—5.83 times upon adding bentonite into the medium. Since the optimal values for phytase activity of most genus Bacillus bacteria are within neutral pH values and temperatures within 50—55°C, a property of B. subtilis IMV B-7023 to show maximum phytase activity at alkaline pH and lower temperatures (28°C) and also stimulation of this activity by soil minerals increases competitiveness of this strain as a component of a bacterial preparation for crop production. Conclusions. Abiotic environmental factors influence the B. subtilis IMV B-7023 supernagrowth and phytase activity. Optimal physicochemical factors for the phytase activity of these bacteria are temperature 28°C and pH 7—12. The concentrations 0.05, 0.5, and 5.0 g/L of silicon dioxide and bentonite increase B. subtilis IMV B-7023 phytase activity. The effect of these nanoscale minerals on the B. subtilis IMV B-7023 growth depends on their type and concentration during cultivation in the medium with phytate as a phosphorus source. The obtained results indicate the potential ability of the B. subtilis IMV B-7023 strain to effectively assimilate phytates in neutral and alkaline soils, especially due to the interaction of these bacteria with bentonite and silicon dioxide nanoparticles. This expands the possibility of using B. subtilis IMV B-7023 in agricultural technologies.
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Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
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