Pub Date : 2024-01-01Epub Date: 2024-01-29DOI: 10.1159/000536491
Yi-Na Park, Hyun Ju Lee, Seung-Ho Ohk
Introduction: The current study investigated the antioxidant and anti-inflammatory effects of ethanol extracts from Lindera glauca twig (LGT) and leaf/stem (LGLS).
Methods: The antioxidant activities were measured by total content of polyphenol and flavonoid, DPPH radical scavenging, and ABTS+ radical scavenging activity. To evaluate the anti-inflammatory effect in the LPS-induced RAW 264.7 cells, protein and mRNA expression of major inflammatory factors were analyzed using Western blot analysis and RT-PCR.
Results: The total polyphenol content of LGT and LGLS was 88.45 ± 11.74 and 115.75 ± 7.87 GA mg/g, respectively. The total flavonoid content was 66 ± 2.89 and 74.33 ± 2.89 QE mg/g. Both LGT and LGLS showed high DPPH and ABTS+ radical scavenging activities. Neither LGT nor LGLS was cytotoxic to RAW 264.7 cells. The anti-inflammatory activities were measured by LPS-induced RAW 264.7 cells. LGT and LGLS showed inhibition of the LPS-induced production of nitric oxide (NO), inducible NO synthase, cyclooxygenase-2 at the protein and mRNA levels, as determined by Western blotting and RT-PCR, respectively. In addition, the release of tumor necrosis factor-α and interleukin-6 mRNA expression levels of these cytokines was reduced by LGT and LGLS.
Conclusion: These results suggest that LGT and LGLS extracts have potential for use as a functional antioxidant and anti-inflammatory ingredient in cosmetic industry.
引言本研究探讨了 Lindera glauca 枝和叶/茎乙醇提取物的抗氧化和抗炎作用:抗氧化活性通过多酚和类黄酮的总含量、DPPH 自由基清除和 ABTS+自由基清除活性进行测定。为了评估在 LPS 诱导的 RAW 264.7 细胞中的抗炎作用,采用 Western 印迹分析和 RT-PCR 分析了主要炎症因子的蛋白质和 mRNA 表达:LGT 和 LGLS 的总多酚含量分别为 88.45±11.74、115.75±7.87 GA mg/g。总黄酮含量分别为 66±2.89、74.33±2.89 QE mg/g。LGT 和 LGLS 均具有较高的 DPPH 和 ABTS+自由基清除活性。LGT 和 LGLS 对 RAW 264.7 细胞均无细胞毒性。抗炎活性是通过 LPS 诱导的 RAW 264.7 细胞进行测定的。经 Western 印迹和 RT-PCR 检测,LGT 和 LGLS 分别在蛋白和 mRNA 水平上抑制了 LPS 诱导的一氧化氮(NO)、诱导型一氧化氮合酶(iNOS)和环氧化酶-2(COX-2)的产生。此外,LGT 和 LGLS 还降低了肿瘤坏死因子-α(TNF- α)和白细胞介素-6(IL-6)的 mRNA 表达水平:这些结果表明,Lindera glauca 树枝和叶/茎提取物具有在化妆品行业用作功能性抗氧化剂和抗炎成分的潜力。
{"title":"Antioxidant and Anti-Inflammatory Activities of Lindera glauca Extracts.","authors":"Yi-Na Park, Hyun Ju Lee, Seung-Ho Ohk","doi":"10.1159/000536491","DOIUrl":"10.1159/000536491","url":null,"abstract":"<p><strong>Introduction: </strong>The current study investigated the antioxidant and anti-inflammatory effects of ethanol extracts from Lindera glauca twig (LGT) and leaf/stem (LGLS).</p><p><strong>Methods: </strong>The antioxidant activities were measured by total content of polyphenol and flavonoid, DPPH radical scavenging, and ABTS+ radical scavenging activity. To evaluate the anti-inflammatory effect in the LPS-induced RAW 264.7 cells, protein and mRNA expression of major inflammatory factors were analyzed using Western blot analysis and RT-PCR.</p><p><strong>Results: </strong>The total polyphenol content of LGT and LGLS was 88.45 ± 11.74 and 115.75 ± 7.87 GA mg/g, respectively. The total flavonoid content was 66 ± 2.89 and 74.33 ± 2.89 QE mg/g. Both LGT and LGLS showed high DPPH and ABTS+ radical scavenging activities. Neither LGT nor LGLS was cytotoxic to RAW 264.7 cells. The anti-inflammatory activities were measured by LPS-induced RAW 264.7 cells. LGT and LGLS showed inhibition of the LPS-induced production of nitric oxide (NO), inducible NO synthase, cyclooxygenase-2 at the protein and mRNA levels, as determined by Western blotting and RT-PCR, respectively. In addition, the release of tumor necrosis factor-α and interleukin-6 mRNA expression levels of these cytokines was reduced by LGT and LGLS.</p><p><strong>Conclusion: </strong>These results suggest that LGT and LGLS extracts have potential for use as a functional antioxidant and anti-inflammatory ingredient in cosmetic industry.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139575970","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-01Epub Date: 2024-01-23DOI: 10.1159/000536327
Urte Clausen, Sören-Tobias Vital, Pia Lambertus, Martina Gehler, Sabine Scheve, Lars Wöhlbrand, Ralf Rabus
Introduction: Phocaeicola vulgatus (formerly Bacteroides vulgatus) is a prevalent member of human and animal guts, where it influences by its dietary-fiber-fueled, fermentative metabolism the microbial community as well as the host health. Moreover, the fermentative metabolism of P. vulgatus bears potential for a sustainable production of bulk chemicals. The aim of the present study was to refine the current understanding of the P. vulgatus physiology.
Methods: P. vulgatus was adapted to anaerobic growth with 14 different carbohydrates, ranging from hexoses, pentoses, hemicellulose, via an uronic acid to deoxy sugars. These substrate-adapted cells formed the basis to define the growth stoichiometries by quantifying growth/fermentation parameters and to reconstruct the catabolic network by applying differential proteomics.
Results: The determination of growth performance revealed, e.g., doubling times (h) from 1.39 (arabinose) to 14.26 (glucuronate), biomass yields (gCDW/mmolS) from 0.01 (fucose) to 0.27 (α-cyclodextrin), and ATP yields (mMATP/mMC) from 0.21 (rhamnose) to 0.60 (glucuronate/xylan). Furthermore, fermentation product spectra were determined, ranging from broad and balanced (with xylan: acetate, succinate, formate, and propanoate) to rather one sided (with rhamnose or fucose: mainly propane-1,2-diol). The fermentation network serving all tested compounds is composed of 56 proteins (all identified), with several peripheral reaction sequences formed with high substrate specificity (e.g., conversion of arabinose to d-xylulose-3-phosphate) implicating a fine-tuned regulation. By contrast, central modules (e.g., glycolysis or the reaction sequence from PEP to succinate) were constitutively formed. Extensive formation of propane-1,2-diol from rhamnose and fucose involves rhamnulokinase (RhaB), rhamnulose-1-phosphate kinase (RhaD), and lactaldehyde reductase (FucO). Furthermore, Sus-like systems are apparently the most relevant uptake systems and a complex array of transmembrane electron-transfer systems (e.g., Na+-pumping Rnf and Nqr complexes, fumarate reductase) as well as F- and V-type ATP-synthases were detected.
Conclusions: The present study provides insights into the potential contribution of P. vulgatus to the gut metabolome and into the strain's biotechnological potential for sustainable production of short-chain fatty acids and alcohols.
Phocaeicola vulgatus(前身为 Bacteroides vulgatus)是人类和动物肠道中的一种常见成员,它以膳食纤维为燃料,通过发酵代谢影响微生物群落和宿主的健康。为了完善目前对 P. vulgatus 生理机能的了解,我们选择了 14 种不同的生长支持碳水化合物(从己糖、戊糖、半纤维素到尿酸再到脱氧糖),以基质适应性细胞为基础,进行了两项主要研究。首先,对生长性能的范围进行了定量评估,结果显示,例如,倍增时间[h]从 1.39(阿拉伯糖)到 14.26(葡萄糖醛酸)不等,生物量产量[gCDW/mmolS]从 0.01(岩藻糖)到 0.27(α-环糊精)不等,ATP 产量[mMATP/mMC]从 0.21(鼠李糖)到 0.60(葡萄糖醛酸/木糖)不等。此外,还测定了发酵产物光谱,其范围从广泛而均衡(木聚糖:乙酸盐、琥珀酸盐、甲酸盐和丙酸盐)到相当片面(鼠李糖或岩藻糖:主要是丙烷-1,2-二醇)不等。其次,根据蛋白质基因组分析,重建了服务于所有测试化合物的发酵网络。该网络由 56 个蛋白质(均已鉴定)组成,其中几个外围反应序列具有高度的底物特异性(如将阿拉伯糖转化为 D-木酮糖-3-磷酸),表明存在微调调节。相比之下,中心模块(如糖酵解或从 PEP 到琥珀酸的反应序列)是组成型形成的。鼠李糖和岩藻糖广泛形成的丙烷-1,2-二醇涉及鼠李糖激酶(RhaB)、鼠李糖-1-磷酸激酶(RhaD)和乳醛还原酶(FucO)。此外,sus-like 系统显然是最相关的吸收系统,还检测到一系列复杂的跨膜电子传递系统(如 Na+ 泵 Rnf 和 Nqr 复合物、富马酸还原酶)以及 F 型和 V 型 ATP 合成酶。综上所述,本研究揭示了P. vulgatus对肠道代谢组的潜在贡献,以及该菌株在可持续生产短链脂肪酸和酒精方面的生物技术潜力。
{"title":"Catabolic Network of the Fermentative Gut Bacterium Phocaeicola vulgatus (Phylum Bacteroidota) from a Physiologic-Proteomic Perspective.","authors":"Urte Clausen, Sören-Tobias Vital, Pia Lambertus, Martina Gehler, Sabine Scheve, Lars Wöhlbrand, Ralf Rabus","doi":"10.1159/000536327","DOIUrl":"10.1159/000536327","url":null,"abstract":"<p><strong>Introduction: </strong>Phocaeicola vulgatus (formerly Bacteroides vulgatus) is a prevalent member of human and animal guts, where it influences by its dietary-fiber-fueled, fermentative metabolism the microbial community as well as the host health. Moreover, the fermentative metabolism of P. vulgatus bears potential for a sustainable production of bulk chemicals. The aim of the present study was to refine the current understanding of the P. vulgatus physiology.</p><p><strong>Methods: </strong>P. vulgatus was adapted to anaerobic growth with 14 different carbohydrates, ranging from hexoses, pentoses, hemicellulose, via an uronic acid to deoxy sugars. These substrate-adapted cells formed the basis to define the growth stoichiometries by quantifying growth/fermentation parameters and to reconstruct the catabolic network by applying differential proteomics.</p><p><strong>Results: </strong>The determination of growth performance revealed, e.g., doubling times (h) from 1.39 (arabinose) to 14.26 (glucuronate), biomass yields (gCDW/mmolS) from 0.01 (fucose) to 0.27 (α-cyclodextrin), and ATP yields (m<sc>M</sc>ATP/m<sc>M</sc>C) from 0.21 (rhamnose) to 0.60 (glucuronate/xylan). Furthermore, fermentation product spectra were determined, ranging from broad and balanced (with xylan: acetate, succinate, formate, and propanoate) to rather one sided (with rhamnose or fucose: mainly propane-1,2-diol). The fermentation network serving all tested compounds is composed of 56 proteins (all identified), with several peripheral reaction sequences formed with high substrate specificity (e.g., conversion of arabinose to <sc>d</sc>-xylulose-3-phosphate) implicating a fine-tuned regulation. By contrast, central modules (e.g., glycolysis or the reaction sequence from PEP to succinate) were constitutively formed. Extensive formation of propane-1,2-diol from rhamnose and fucose involves rhamnulokinase (RhaB), rhamnulose-1-phosphate kinase (RhaD), and lactaldehyde reductase (FucO). Furthermore, Sus-like systems are apparently the most relevant uptake systems and a complex array of transmembrane electron-transfer systems (e.g., Na+-pumping Rnf and Nqr complexes, fumarate reductase) as well as F- and V-type ATP-synthases were detected.</p><p><strong>Conclusions: </strong>The present study provides insights into the potential contribution of P. vulgatus to the gut metabolome and into the strain's biotechnological potential for sustainable production of short-chain fatty acids and alcohols.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139542428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2023-01-10DOI: 10.1159/000529038
Alaska Pokhrel, Hue Dinh, Liping Li, Karl A Hassan, Amy K Cain, Ian T Paulsen
l-cysteine biosynthesis from inorganic sulfur represents a major mechanism by which reduced sulfur is incorporated into organic compounds. Cysteine biosynthesis and regulation is well characterized in Escherichia coli. However, the regulation of sulfur metabolism in Acinetobacter baumannii is only partly understood, with the LysR-type regulator, GigC known to control some aspects of sulfur reduction. In this study, we have used transcriptomics and bioinformatic analyses to characterize a novel LysR-type transcriptional regulator encoded by ABUW_1016 (cbl), in a highly multidrug resistant and virulent isolate of A. baumannii. We have shown that Cbl is involved in controlling expression of the genes required for uptake and reduction of various sulfur sources in A. baumannii. Collectively, we have identified the global regulon of Cbl and proposed a model of cysteine biosynthesis and its regulation by Cbl and GigC in A. baumannii.
{"title":"Identification of a Novel LysR Family Transcriptional Regulator Controlling Acquisition of Sulfur Sources in Acinetobacter baumannii.","authors":"Alaska Pokhrel, Hue Dinh, Liping Li, Karl A Hassan, Amy K Cain, Ian T Paulsen","doi":"10.1159/000529038","DOIUrl":"10.1159/000529038","url":null,"abstract":"<p><p><sc>l</sc>-cysteine biosynthesis from inorganic sulfur represents a major mechanism by which reduced sulfur is incorporated into organic compounds. Cysteine biosynthesis and regulation is well characterized in Escherichia coli. However, the regulation of sulfur metabolism in Acinetobacter baumannii is only partly understood, with the LysR-type regulator, GigC known to control some aspects of sulfur reduction. In this study, we have used transcriptomics and bioinformatic analyses to characterize a novel LysR-type transcriptional regulator encoded by ABUW_1016 (cbl), in a highly multidrug resistant and virulent isolate of A. baumannii. We have shown that Cbl is involved in controlling expression of the genes required for uptake and reduction of various sulfur sources in A. baumannii. Collectively, we have identified the global regulon of Cbl and proposed a model of cysteine biosynthesis and its regulation by Cbl and GigC in A. baumannii.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10515810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The cultural parameters of Streptomyces sp. for pectinase production were optimized using the Box-Behnken design. The maximum pectinase production was obtained after 58 h at 35°C and pH 7 upon submerged fermentation in yeast extract-containing media. The enzymes were partially purified with acetone precipitation, and the analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram revealed that Streptomyces sp. produced two pectinases protein with molecular weights of about 25 and 75 kDa. The pectinase activity was detected in a wide range of temperatures (30°C-80°C) and pH (3-9) with maximum pectinase activities observed at 70°C and pH 5 and 9. The enzymes retained about 30-40% of their activities even after incubating the enzyme at different temperatures for 120 min. The pectinase activities of Streptomyces sp. were enhanced in the media containing 1.5% pectin, 1% casein as a nitrogen source, 0.5 mM MgSO4, and 5 mM NaCl. Further, the addition of Tween-20, amino acids, and vitamins to the media also enhanced the pectinase activity. Moreover, the bacterium illustrated the ability to decolorize crystal violet dye efficiently. The decolorization rate ranged from 39.29 to 53.75%, showing the highest bacterial decolorization in the media containing 2 mg/mL crystal violet at 144 h. Therefore, the bacterium has the potential in treating wastewater produced by industries like textile industries.
{"title":"Optimization of Cultural Conditions for Pectinase Production by Streptomyces sp. and Characterization of Partially Purified Enzymes.","authors":"Sarita Shrestha, Chonlong Chio, Janak Raj Khatiwada, Aristide Laurel Mokale Kognou, Xuantong Chen, Wensheng Qin","doi":"10.1159/000528257","DOIUrl":"10.1159/000528257","url":null,"abstract":"<p><p>The cultural parameters of Streptomyces sp. for pectinase production were optimized using the Box-Behnken design. The maximum pectinase production was obtained after 58 h at 35°C and pH 7 upon submerged fermentation in yeast extract-containing media. The enzymes were partially purified with acetone precipitation, and the analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram revealed that Streptomyces sp. produced two pectinases protein with molecular weights of about 25 and 75 kDa. The pectinase activity was detected in a wide range of temperatures (30°C-80°C) and pH (3-9) with maximum pectinase activities observed at 70°C and pH 5 and 9. The enzymes retained about 30-40% of their activities even after incubating the enzyme at different temperatures for 120 min. The pectinase activities of Streptomyces sp. were enhanced in the media containing 1.5% pectin, 1% casein as a nitrogen source, 0.5 mM MgSO4, and 5 mM NaCl. Further, the addition of Tween-20, amino acids, and vitamins to the media also enhanced the pectinase activity. Moreover, the bacterium illustrated the ability to decolorize crystal violet dye efficiently. The decolorization rate ranged from 39.29 to 53.75%, showing the highest bacterial decolorization in the media containing 2 mg/mL crystal violet at 144 h. Therefore, the bacterium has the potential in treating wastewater produced by industries like textile industries.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40701873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2022-08-30DOI: 10.1159/000526662
Jennie C Hildenbrand, Georg A Sprenger, Attila Teleki, Ralf Takors, Dieter Jendrossek
Polyphosphate kinases (PPKs) catalyze the reversible transfer of the γ-phosphate moiety of ATP (or of another nucleoside triphosphate) to a growing chain of polyphosphate (polyP). In this study, we describe that PPKs of various sources are additionally able to phosphorylate thiamine diphosphate (ThP2) to produce thiamine triphosphate (ThP3) and even thiamine tetraphosphate in vitro using polyP as phosphate donor. Furthermore, all tested PPK2s, but not PPK1s, were able to phosphorylate thiamine monophosphate (ThP1) to ThP2 and ThP3 although at low efficiency. The predicted masses and identities of the mono- and oligo-phosphorylated thiamine metabolites were identified by high-performance liquid chromatography tandem mass spectrometry. Moreover, the biological activity of ThP2, that was synthesized by phosphorylation of ThP1 with polyP and PPK, as a cofactor of ThP2-dependent enzymes (here transketolase TktA from Escherichia coli) was confirmed in a coupled enzyme assay. Our study shows that PPKs are promiscuous enzymes in vitro that could be involved in the formation of a variety of phosphorylated metabolites in vivo.
{"title":"Polyphosphate Kinases Phosphorylate Thiamine Phosphates.","authors":"Jennie C Hildenbrand, Georg A Sprenger, Attila Teleki, Ralf Takors, Dieter Jendrossek","doi":"10.1159/000526662","DOIUrl":"10.1159/000526662","url":null,"abstract":"<p><p>Polyphosphate kinases (PPKs) catalyze the reversible transfer of the γ-phosphate moiety of ATP (or of another nucleoside triphosphate) to a growing chain of polyphosphate (polyP). In this study, we describe that PPKs of various sources are additionally able to phosphorylate thiamine diphosphate (ThP2) to produce thiamine triphosphate (ThP3) and even thiamine tetraphosphate in vitro using polyP as phosphate donor. Furthermore, all tested PPK2s, but not PPK1s, were able to phosphorylate thiamine monophosphate (ThP1) to ThP2 and ThP3 although at low efficiency. The predicted masses and identities of the mono- and oligo-phosphorylated thiamine metabolites were identified by high-performance liquid chromatography tandem mass spectrometry. Moreover, the biological activity of ThP2, that was synthesized by phosphorylation of ThP1 with polyP and PPK, as a cofactor of ThP2-dependent enzymes (here transketolase TktA from Escherichia coli) was confirmed in a coupled enzyme assay. Our study shows that PPKs are promiscuous enzymes in vitro that could be involved in the formation of a variety of phosphorylated metabolites in vivo.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40331484","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2023-09-30DOI: 10.1159/000532088
Gunnar Sturm, Mohammad Mojarrad, Anne-Kristin Kaster
To date, the vast majority of prokaryotic organisms escapes detailed characterization because they cannot be isolated in axenic cultures. These organisms are referred to as microbial dark matter. Targeted labelling and sorting of these microorganisms pave the way for single-cell, enrichment, or cultivation approaches. In this review, we describe an array of different methods ranging from labeling-free to specific labelling techniques. In addition, different cell sorting methods and their combinations with targeting strategies are summarized and downstream applications like sequencing and cultivation are reviewed. Recent advances, challenges, and limitations of the particular methods are discussed with respect to cell viability, genome integrity as well as throughput, in order to help researchers select the most suitable methods for their specific research questions.
{"title":"Targeted Cell Labeling and Sorting of Prokaryotes for Cultivation and Omics Approaches.","authors":"Gunnar Sturm, Mohammad Mojarrad, Anne-Kristin Kaster","doi":"10.1159/000532088","DOIUrl":"10.1159/000532088","url":null,"abstract":"<p><p>To date, the vast majority of prokaryotic organisms escapes detailed characterization because they cannot be isolated in axenic cultures. These organisms are referred to as microbial dark matter. Targeted labelling and sorting of these microorganisms pave the way for single-cell, enrichment, or cultivation approaches. In this review, we describe an array of different methods ranging from labeling-free to specific labelling techniques. In addition, different cell sorting methods and their combinations with targeting strategies are summarized and downstream applications like sequencing and cultivation are reviewed. Recent advances, challenges, and limitations of the particular methods are discussed with respect to cell viability, genome integrity as well as throughput, in order to help researchers select the most suitable methods for their specific research questions.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41141966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2023-06-15DOI: 10.1159/000531468
Kevin J Hendargo, Ashay O Patel, Onyeka S Chukwudozie, Gabriel Moreno-Hagelsieb, J Andres Christen, Arturo Medrano-Soto, Milton H Saier
Members of the Piezo family of mechanically activated cation channels are involved in multiple physiological processes in higher eukaryotes, including vascular development, cell differentiation, touch perception, hearing, and more, but they are also common in single-celled eukaryotic microorganisms. Mutations in these proteins in humans are associated with a variety of diseases, such as colorectal adenomatous polyposis, dehydrated hereditary stomatocytosis, and hereditary xerocytosis. Available 3D structures for Piezo proteins show nine regions of four transmembrane segments each that have the same fold. Despite the remarkable similarity among the nine characteristic structural repeats in the family, no significant sequence similarity among them has been reported. Using bioinformatics approaches and the Transporter Classification Database (TCDB) as reference, we reliably identified sequence similarity among repeats based on four lines of evidence: (1) hidden Markov model-profile similarities across repeats at the family level, (2) pairwise sequence similarities between different repeats across Piezo homologs, (3) Piezo-specific conserved sequence signatures that consistently identify the same regions across repeats, and (4) conserved residues that maintain the same orientation and location in 3D space.
{"title":"Sequence Similarity among Structural Repeats in the Piezo Family of Mechanosensitive Ion Channels.","authors":"Kevin J Hendargo, Ashay O Patel, Onyeka S Chukwudozie, Gabriel Moreno-Hagelsieb, J Andres Christen, Arturo Medrano-Soto, Milton H Saier","doi":"10.1159/000531468","DOIUrl":"10.1159/000531468","url":null,"abstract":"<p><p>Members of the Piezo family of mechanically activated cation channels are involved in multiple physiological processes in higher eukaryotes, including vascular development, cell differentiation, touch perception, hearing, and more, but they are also common in single-celled eukaryotic microorganisms. Mutations in these proteins in humans are associated with a variety of diseases, such as colorectal adenomatous polyposis, dehydrated hereditary stomatocytosis, and hereditary xerocytosis. Available 3D structures for Piezo proteins show nine regions of four transmembrane segments each that have the same fold. Despite the remarkable similarity among the nine characteristic structural repeats in the family, no significant sequence similarity among them has been reported. Using bioinformatics approaches and the Transporter Classification Database (TCDB) as reference, we reliably identified sequence similarity among repeats based on four lines of evidence: (1) hidden Markov model-profile similarities across repeats at the family level, (2) pairwise sequence similarities between different repeats across Piezo homologs, (3) Piezo-specific conserved sequence signatures that consistently identify the same regions across repeats, and (4) conserved residues that maintain the same orientation and location in 3D space.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":0.1,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11283329/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9639934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2023-03-21DOI: 10.1159/000530228
Aristide Laurel Mokale Kognou, Chonlong Chio, Janak Raj Khatiwada, Sarita Shrestha, Xuantong Chen, Yuen Zhu, Rosalie Anne Ngono Ngane, Gabriel Agbor Agbor, Zi-Hua Jiang, Chunbao Charles Xu, Wensheng Qin
Soil bacteria participate in self-immobilization processes for survival, persistence, and production of virulence factors in some niches or hosts through their capacities for autoaggregation, cell surface hydrophobicity, biofilm formation, and antibiotic and heavy metal resistance. This study investigated potential virulence, antibiotic and heavy metal resistance, solvent adhesion, and biofilm-forming capabilities of six cellulolytic bacteria isolated from soil samples: Paenarthrobacter sp. MKAL1, Hymenobacter sp. MKAL2, Mycobacterium sp. MKAL3, Stenotrophomonas sp. MKAL4, Chryseobacterium sp. MKAL5, and Bacillus sp. MKAL6. Strains were subjected to phenotypic methods, including heavy metal and antibiotic susceptibility and virulence factors (protease, lipase, capsule production, autoaggregation, hydrophobicity, and biofilm formation). The effect of ciprofloxacin was also investigated on bacterial susceptibility over time, cell membrane, and biofilm formation. Strains MKAL2, MKAL5, and MKAL6 exhibited protease and lipase activities, while only MKAL6 produced capsules. All strains were capable of aggregating, forming biofilm, and adhering to solvents. Strains tolerated high amounts of chromium, lead, zinc, nickel, and manganese and were resistant to lincomycin. Ciprofloxacin exhibited bactericidal activity against these strains. Although the phenotypic evaluation of virulence factors of bacteria can indicate their pathogenic nature, an in-depth genetic study of virulence, antibiotic and heavy metal resistance genes is required.
{"title":"Characterization of Potential Virulence, Resistance to Antibiotics and Heavy Metals, and Biofilm-Forming Capabilities of Soil Lignocellulolytic Bacteria.","authors":"Aristide Laurel Mokale Kognou, Chonlong Chio, Janak Raj Khatiwada, Sarita Shrestha, Xuantong Chen, Yuen Zhu, Rosalie Anne Ngono Ngane, Gabriel Agbor Agbor, Zi-Hua Jiang, Chunbao Charles Xu, Wensheng Qin","doi":"10.1159/000530228","DOIUrl":"10.1159/000530228","url":null,"abstract":"<p><p>Soil bacteria participate in self-immobilization processes for survival, persistence, and production of virulence factors in some niches or hosts through their capacities for autoaggregation, cell surface hydrophobicity, biofilm formation, and antibiotic and heavy metal resistance. This study investigated potential virulence, antibiotic and heavy metal resistance, solvent adhesion, and biofilm-forming capabilities of six cellulolytic bacteria isolated from soil samples: Paenarthrobacter sp. MKAL1, Hymenobacter sp. MKAL2, Mycobacterium sp. MKAL3, Stenotrophomonas sp. MKAL4, Chryseobacterium sp. MKAL5, and Bacillus sp. MKAL6. Strains were subjected to phenotypic methods, including heavy metal and antibiotic susceptibility and virulence factors (protease, lipase, capsule production, autoaggregation, hydrophobicity, and biofilm formation). The effect of ciprofloxacin was also investigated on bacterial susceptibility over time, cell membrane, and biofilm formation. Strains MKAL2, MKAL5, and MKAL6 exhibited protease and lipase activities, while only MKAL6 produced capsules. All strains were capable of aggregating, forming biofilm, and adhering to solvents. Strains tolerated high amounts of chromium, lead, zinc, nickel, and manganese and were resistant to lincomycin. Ciprofloxacin exhibited bactericidal activity against these strains. Although the phenotypic evaluation of virulence factors of bacteria can indicate their pathogenic nature, an in-depth genetic study of virulence, antibiotic and heavy metal resistance genes is required.</p>","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9156717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhaoyang Liu, Bingcong Ji, Yaqing Zhang, Xunli Liu
Jujube is an important economic crop in the Xinjiang Uygur Autonomous Region. Microbial diversity in the rhizosphere is essential for plant quality; however, soil bacterial diversity and community structure in the jujube rhizosphere have not been characterized in this region. In this study, we used pyrosequencing to analyze bacterial diversity and community structure at different growth stages in the jujube rhizosphere in Hetian, Kashi, and Aksu prefectures. These results revealed a greater bacterial diversity in the 8-year jujube rhizosphere as compared with the 3-year-old rhizosphere taken from the same sampling area. Moreover, samples obtained from Kashi prefecture showed the largest diversity among the different areas. The most abundant phyla across all soil samples were Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, and Firmicutes. Dominant phyla in the 8-year jujube rhizosphere accounted for the increased observed diversity. Furthermore, comparative analysis of the bacterial communities with respect to rhizosphere age and sampling areas revealed a significant correlation between soil properties and phyla diversity. To the best of our knowledge, this is the first study of jujube rhizosphere bacterial diversity and community structure in the southern Xinjiang Uygur Autonomous Region, and we hope that our research provides a reference for future studies.
{"title":"Bacterial Diversity and Community Structure of the Jujube Rhizosphere in Southern Xinjiang Uygur Autonomous Region, China","authors":"Zhaoyang Liu, Bingcong Ji, Yaqing Zhang, Xunli Liu","doi":"10.1159/000525000","DOIUrl":"https://doi.org/10.1159/000525000","url":null,"abstract":"Jujube is an important economic crop in the Xinjiang Uygur Autonomous Region. Microbial diversity in the rhizosphere is essential for plant quality; however, soil bacterial diversity and community structure in the jujube rhizosphere have not been characterized in this region. In this study, we used pyrosequencing to analyze bacterial diversity and community structure at different growth stages in the jujube rhizosphere in Hetian, Kashi, and Aksu prefectures. These results revealed a greater bacterial diversity in the 8-year jujube rhizosphere as compared with the 3-year-old rhizosphere taken from the same sampling area. Moreover, samples obtained from Kashi prefecture showed the largest diversity among the different areas. The most abundant phyla across all soil samples were Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, and Firmicutes. Dominant phyla in the 8-year jujube rhizosphere accounted for the increased observed diversity. Furthermore, comparative analysis of the bacterial communities with respect to rhizosphere age and sampling areas revealed a significant correlation between soil properties and phyla diversity. To the best of our knowledge, this is the first study of jujube rhizosphere bacterial diversity and community structure in the southern Xinjiang Uygur Autonomous Region, and we hope that our research provides a reference for future studies.","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2022-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42646204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Arne Weiten, K. Kalvelage, Meina Neumann-Schaal, Ramona Buschen, Sabine Scheve, M. Winklhofer, R. Rabus
Phaeobacter inhibens DSM 17395 is a heterotrophic member of the ubiquitous, marine Roseobacter group and specializes in the aerobic utilization of carbohydrates and amino acids via pathways widespread among roseobacters. The in vivo responsiveness of P. inhibens DSM 17395 was studied with nonadapted cells (succinate-grown), which were exposed to a single pulse (100–0.01 µM) each of N-acetylglucosamine, mannitol, xylose, leucine, phenylalanine, or tryptophan (effectors). Responsiveness was then determined by time-resolved transcript analyses (quantitative reverse transcription-PCR) of “degradation” and “uptake” genes selected based on previously reported substrate-specific proteome profiles. The transcriptional response thresholds were: 50–100 nM for nagK (N-acetylglucosamine kinase), paaA (ring 1,2-phenylacetyl-CoA epoxidase), and kynA (tryptophan 2,3-dioxygenase), 10–50 nM for xylA (xylose isomerase), and around 10 nM for mtlK (mannitol 2-dehydrogenase). A threshold for leucine could not be determined due to the elevated intrinsic presence of leucine in the exometabolome of succinate-grown cells (no effector addition). Notably, the response thresholds for presumptive carbohydrate-binding proteins of ABC-transporters were in the same range or even lower: 0.1–1 µM for c27930 (N-acetylglucosamine) and even below 10 nM for c13210 (mannitol) and xylF (xylose). These results shed new light on the sensory/regulatory sensitivity of a well-studied roseobacter for recognizing potential substrates at low ambient concentrations and on the concentration threshold below which these might escape biodegradation (“emergent recalcitrance” concept of dissolved organic matter persistence).
{"title":"Nanomolar Responsiveness of Marine Phaeobacter inhibens DSM 17395 toward Carbohydrates and Amino Acids","authors":"Arne Weiten, K. Kalvelage, Meina Neumann-Schaal, Ramona Buschen, Sabine Scheve, M. Winklhofer, R. Rabus","doi":"10.1159/000524702","DOIUrl":"https://doi.org/10.1159/000524702","url":null,"abstract":"Phaeobacter inhibens DSM 17395 is a heterotrophic member of the ubiquitous, marine Roseobacter group and specializes in the aerobic utilization of carbohydrates and amino acids via pathways widespread among roseobacters. The in vivo responsiveness of P. inhibens DSM 17395 was studied with nonadapted cells (succinate-grown), which were exposed to a single pulse (100–0.01 µM) each of N-acetylglucosamine, mannitol, xylose, leucine, phenylalanine, or tryptophan (effectors). Responsiveness was then determined by time-resolved transcript analyses (quantitative reverse transcription-PCR) of “degradation” and “uptake” genes selected based on previously reported substrate-specific proteome profiles. The transcriptional response thresholds were: 50–100 nM for nagK (N-acetylglucosamine kinase), paaA (ring 1,2-phenylacetyl-CoA epoxidase), and kynA (tryptophan 2,3-dioxygenase), 10–50 nM for xylA (xylose isomerase), and around 10 nM for mtlK (mannitol 2-dehydrogenase). A threshold for leucine could not be determined due to the elevated intrinsic presence of leucine in the exometabolome of succinate-grown cells (no effector addition). Notably, the response thresholds for presumptive carbohydrate-binding proteins of ABC-transporters were in the same range or even lower: 0.1–1 µM for c27930 (N-acetylglucosamine) and even below 10 nM for c13210 (mannitol) and xylF (xylose). These results shed new light on the sensory/regulatory sensitivity of a well-studied roseobacter for recognizing potential substrates at low ambient concentrations and on the concentration threshold below which these might escape biodegradation (“emergent recalcitrance” concept of dissolved organic matter persistence).","PeriodicalId":18457,"journal":{"name":"Microbial Physiology","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2022-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49384769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}