holosiivsky国家自然公园化学有机营养细菌的生理生态特性和水解活性

Q4 Biochemistry, Genetics and Molecular Biology
G. Gladka, N. Borzova, O. Gudzenko, V. Hovorukha, О. Havryliuk, O. Shabliĭ, L. Yastremska, O. Tashyrev
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Meat-peptone agar was used to cultivate bacteria. Bacterial UV irradiation was performed with a BUF-15 lamp (λ=254 nm) in the range of 30—1350 J/m2. The temperature range of growth and halotolerance of microorganisms was determined in the range of 1—42°C and 0.1—150 g NaCl/L, respectively. Bacterial isolates were cultivated in submerged conditions at 28°C for 4 days. Synthetic p-nitrophenyl substrates, soluble starch, and guar galactomannan were used to determine glycosidase activity. To study proteolytic activity, casein, elastin, and gelatin were used. Results. The study of 14 soil and plant samples revealed the number of bacteria detected from 9.3×104 to 4.8×105 CFU/g in winter, and 4.8×105 to 4.2×106 CFU/g in summer. The microorganisms were represented by 1—4 morphotypes. There were isolated 37 isolates of aerobic chemoorganotrophic microorganisms, and 69% of them were represented by gram-positive rods. Th e dominance of pigmented isolates was not detected. 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引用次数: 0

摘要

任何自然生态系统都包含一定范围的微生物。人为影响可引起土壤和根际微生物群生长条件的变化,影响微生物的数量和生理特性。本研究旨在分离乌克兰Holosiivskyi国家自然公园(Holosiivskyi National Nature Park, Ukraine)陆地生态系统中未暴露于极端因素的代表性微生物,研究其生态生理特性(抗紫外线辐射、脱水、高盐、高温),并研究其胞外糖苷和蛋白水解活性。方法。研究了在30°C条件下从Holosiivskyi国家自然公园土壤和植物群落中分离出的好氧化能有机营养细菌。采用肉蛋白胨琼脂培养细菌。采用BUF-15灯(λ=254 nm)进行细菌紫外照射,照射范围为30-1350 J/m2。测定微生物的生长温度范围为1 ~ 42℃,耐盐温度范围为0.1 ~ 150 g NaCl/L。分离的细菌在28°C的水浸条件下培养4天。采用合成对硝基苯底物、可溶性淀粉和瓜尔胶半乳甘露聚糖测定糖苷酶活性。用酪蛋白、弹性蛋白和明胶研究蛋白水解活性。结果。对14份土壤和植物样品的研究表明,冬季检测到的细菌数量为9.3×104 ~ 4.8×105 CFU/g,夏季为4.8×105 ~ 4.2×106 CFU/g。微生物有1-4种形态类型。共分离到37株好氧化有机营养微生物,其中69%为革兰氏阳性杆状体。未检测到色素分离株的优势性。研究的大多数微生物是耐寒和中等嗜盐菌。从黑土和深灰色土壤中分离得到的3g3、8g1、8g2、8g3对紫外辐射具有较高的抗性。LD99.99范围为800 ~ 1100 J/m2。褐藓、桦树苔、沙土绿苔和橡树苔(1g、4g2、9g1、14g2)的分离株具有中等抗性。LD99.99为280 ~ 650 J/m2。抗紫外线能力与色素沉着无关。它与脱水有关。对这种紫外线辐射和脱水的抵抗现象可能表明存在DNA损伤的主动修复机制。所有菌株均具有纤维素和半纤维素降解活性以及酪蛋白水解活性。分离物9g1表现出较高的β-木糖苷酶活性。结论。非适应微生物对紫外线辐射和脱水的高抗性以及广泛的外水解酶活性表明,自然生态系统中微生物具有广泛的适应能力,超越了周围因素的影响。在现有文献中没有数据存在定义水解活性和乌克兰温带地区的微生物对极端因素的抗性。获得的实验数据将有助于更好地了解温带地区微生物对极端因素的抗性水平。结果,分离出了具有高降解活性的新细菌。所研究的分离物需要进一步的表征和分析以用于生物技术应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Ecophysiological Properties and Hydrolytic Activity of Chemoorganotrophic Bacteria from Holosiivskyi National Nature Park
Any natural ecosystem contains a specific range of microorganisms. The anthropogenic impact can cause a change in the growth conditions of soil and rhizospheric microbiome and affect the number and the physiological properties of microorganisms. The aim of the study was to isolate the representative microorganisms from terrestrial ecosystems of Holosiivskyi National Nature Park (Ukraine) that are not exposed to extreme factors, to study their ecophysiological properties (resistance to UV radiation, dehydration, hypersalinity, temperature), and to study the extracellular glycoside and proteolytic activities. Methods. Aerobic chemoorganotrophic bacteria isolated at 30°C from soil and phytocenoses of Holosiivskyi National Nature Park were studied. Meat-peptone agar was used to cultivate bacteria. Bacterial UV irradiation was performed with a BUF-15 lamp (λ=254 nm) in the range of 30—1350 J/m2. The temperature range of growth and halotolerance of microorganisms was determined in the range of 1—42°C and 0.1—150 g NaCl/L, respectively. Bacterial isolates were cultivated in submerged conditions at 28°C for 4 days. Synthetic p-nitrophenyl substrates, soluble starch, and guar galactomannan were used to determine glycosidase activity. To study proteolytic activity, casein, elastin, and gelatin were used. Results. The study of 14 soil and plant samples revealed the number of bacteria detected from 9.3×104 to 4.8×105 CFU/g in winter, and 4.8×105 to 4.2×106 CFU/g in summer. The microorganisms were represented by 1—4 morphotypes. There were isolated 37 isolates of aerobic chemoorganotrophic microorganisms, and 69% of them were represented by gram-positive rods. Th e dominance of pigmented isolates was not detected. Most of the microorganisms studied were psychrotolerant and moderate halophiles. The isolates 3g3, 8g1, 8g2, 8g3 from chornozem and dark gray soil showed high resistance to UV radiation. Th e LD99.99 ranged from 800 to 1100 J/m2. The isolates from chornozem, birch moss, green moss with sand and soil, and green moss from oak (1g, 4g2, 9g1, 14g2) were moderately resistant. The LD99.99 was 280—650 J/m2. The UV resistance was shown to be independent of pigmentation. It correlated with dehydration. The phenomenon of resistance to such UV radiation and dehydration may indicate the presence of active reparation mechanisms of DNA damage. All isolates showed cellulose and hemicellulose degrading activities as well as caseinolytic activity. Isolate 9g1 showed high β-xylosidase activity. Conclusions. The high resistance to UV radiation and dehydration of non-adapted microorganisms as well as the wide range of exohydrolase activity indicate the wide adaptive capacity of microorganisms from natural ecosystems, which goes beyond the influence of surrounding factors. No data existed in the available literature defining hydrolytic activity and resistance of microorganisms of the temperate region of Ukraine to extreme factors. The obtained experimental data will allow for a better understanding of the resistance level of microorganisms of temperate regions to extreme factors. As a result of the work, new bacteria with high degrading activity were isolated. The studied isolates require further characterization and analysis for biotechnological applications.
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Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
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