ExtremophilesPub Date : 2023-07-23DOI: 10.1007/s00792-023-01304-4
Michael T Madigan, Kelly S Bender, Sophia A Sanguedolce, Mary N Parenteau, Marisa H Mayer, Yukihiro Kimura, Zheng-Yu Wang-Otomo, W Matthew Sattley
{"title":"Genomic basis for the unique phenotype of the alkaliphilic purple nonsulfur bacterium Rhodobaca bogoriensis.","authors":"Michael T Madigan, Kelly S Bender, Sophia A Sanguedolce, Mary N Parenteau, Marisa H Mayer, Yukihiro Kimura, Zheng-Yu Wang-Otomo, W Matthew Sattley","doi":"10.1007/s00792-023-01304-4","DOIUrl":"10.1007/s00792-023-01304-4","url":null,"abstract":"<p><p>Although several species of purple sulfur bacteria inhabit soda lakes, Rhodobaca bogoriensis is the first purple nonsulfur bacterium cultured from such highly alkaline environments. Rhodobaca bogoriensis strain LBB1<sup>T</sup> was isolated from Lake Bogoria, a soda lake in the African Rift Valley. The phenotype of Rhodobaca bogoriensis is unique among purple bacteria; the organism is alkaliphilic but not halophilic, produces carotenoids absent from other purple nonsulfur bacteria, and is unable to grow autotrophically or fix molecular nitrogen. Here we analyze the draft genome sequence of Rhodobaca bogoriensis to gain further insight into the biology of this extremophilic purple bacterium. The strain LBB1<sup>T</sup> genome consists of 3.91 Mbp with no plasmids. The genome sequence supports the defining characteristics of strain LBB1<sup>T</sup>, including its (1) production of a light-harvesting 1-reaction center (LH1-RC) complex but lack of a peripheral (LH2) complex, (2) ability to synthesize unusual carotenoids, (3) capacity for both phototrophic (anoxic/light) and chemotrophic (oxic/dark) energy metabolisms, (4) utilization of a wide variety of organic compounds (including acetate in the absence of a glyoxylate cycle), (5) ability to oxidize both sulfide and thiosulfate despite lacking the capacity for autotrophic growth, and (6) absence of a functional nitrogen-fixation system for diazotrophic growth. The assortment of properties in Rhodobaca bogoriensis has no precedent among phototrophic purple bacteria, and the results are discussed in relation to the organism's soda lake habitat and evolutionary history.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"19"},"PeriodicalIF":2.9,"publicationDate":"2023-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9877045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-07-23DOI: 10.1007/s00792-023-01306-2
Aveepsa Sengupta, Kunwali Das, Nidhi Jha, Yusuf Akhter, Ashutosh Kumar
{"title":"Molecular evolution steered structural adaptations in the DNA polymerase III α subunit of halophilic bacterium Salinibacter ruber.","authors":"Aveepsa Sengupta, Kunwali Das, Nidhi Jha, Yusuf Akhter, Ashutosh Kumar","doi":"10.1007/s00792-023-01306-2","DOIUrl":"https://doi.org/10.1007/s00792-023-01306-2","url":null,"abstract":"<p><p>A significant portion of the earth has a salty environment, and the literature on bacterial survival mechanisms in salty environments is limited. During molecular evolution, halophiles increase acidic amino acid residues on their protein surfaces which leads to a negatively charged surface potential that helps them to maintain the protein integrity and protect them from denaturation by competing with salt ions. Through protein family analysis, we have investigated the molecular-level adaptive features of DNA polymerase III's catalytic subunit (alpha) and its structure-function relationship. This study throws light on the novel understanding of halophilic bacterial replication and the molecular basis of salt adaptation. Comparisons of the amino acid contents and electronegativity of halophilic and mesophilic bacterial proteins revealed adaptations that allow halophilic bacteria to thrive in high salt concentrations. A significantly lower isoelectric point of halophilic bacterial proteins indicates the acidic nature. Also, an abundance of disordered regions in halophiles suggests the requirement of the salt ions that play a crucial role in their stable protein folding. Despite having similar topology, mesophilic and halophilic proteins, a set of very prominent molecular modifications was observed in the alpha subunit of halophiles.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"20"},"PeriodicalIF":2.9,"publicationDate":"2023-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9877046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-07-10DOI: 10.1007/s00792-023-01299-y
Rodrigo Cortés-Antiquera, Sebastián L Márquez, Giannina Espina, Jorge Sánchez-SanMartín, Jenny M Blamey
{"title":"Recombinant expression and characterization of a new laccase, bioinformatically identified, from the Antarctic thermophilic bacterium Geobacillus sp. ID17.","authors":"Rodrigo Cortés-Antiquera, Sebastián L Márquez, Giannina Espina, Jorge Sánchez-SanMartín, Jenny M Blamey","doi":"10.1007/s00792-023-01299-y","DOIUrl":"https://doi.org/10.1007/s00792-023-01299-y","url":null,"abstract":"<p><p>Geobacillus sp. ID17 is a gram-positive thermophilic bacterium isolated from Deception Island, Antarctica, which has shown to exhibit remarkable laccase activity in crude extract at high temperatures. A bioinformatic search using local databases led to the identification of three putative multicopper oxidase sequences in the genome of this microorganism. Sequence analysis revealed that one of those sequences contains the four-essential copper-binding sites present in other well characterized laccases. The gene encoding this sequence was cloned and overexpressed in Escherichia coli, partially purified and preliminary biochemically characterized. The resulting recombinant enzyme was recovered in active and soluble form, exhibiting optimum copper-dependent laccase activity at 55 °C, pH 6.5 with syringaldazine substrate, retaining over 60% of its activity after 1 h at 55 and 60 °C. In addition, this thermophilic enzyme is not affected by common inhibitors SDS, NaCl and L-cysteine. Furthermore, biodecolorization assays revealed that this laccase is capable of degrading 60% of malachite green, 54% of Congo red, and 52% of Remazol Brilliant Blue R, after 6 h at 55 °C with aid of ABTS as redox mediator. The observed properties of this enzyme and the relatively straightforward overexpression and partial purification of it could be of great interest for future biotechnology applications.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"18"},"PeriodicalIF":2.9,"publicationDate":"2023-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9866583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-07-07DOI: 10.1007/s00792-023-01302-6
Aurélien Daussin, Pauline Vannier, Émilien Mater, Tina Šantl-Temkiv, Charles Cockell, Viggó Þór Marteinsson
{"title":"Survival of Icelandic airborne microbes towards simulated atmospheric stress factors.","authors":"Aurélien Daussin, Pauline Vannier, Émilien Mater, Tina Šantl-Temkiv, Charles Cockell, Viggó Þór Marteinsson","doi":"10.1007/s00792-023-01302-6","DOIUrl":"https://doi.org/10.1007/s00792-023-01302-6","url":null,"abstract":"<p><p>Surface microbes are aerosolized into the atmosphere by wind and events such as dust storms, wildland fires, and volcano eruptions. Only microbial cells that survive the various atmospheric stressors during their transportation will deposit and colonize new environments. These stressors include desiccation, oxidative stress, solar radiation, osmotic shock, and freeze-thaw cycles. In this paper, we specifically studied the survival of representative microbial model strains isolated from the atmosphere over pristine volcanic landscapes to understand their potential to successfully disperse to novel terrestrial environments. In line with previous studies, we found that the most stringent selection factors were the freeze-thaw and osmotic shock cycles and that the strains affiliated with Proteobacteria and Ascomycota were the best to survive simulated atmospheric stresses. Specifically, isolates belonging to Paracoccus marinus, Janthinobacterium rivuli, and Sarocladium kiliense exhibited the highest levels of resistance to atmospheric stress. However, the number of strains tested in our study was limited and caution should be taken when generalizing these findings.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"17"},"PeriodicalIF":2.9,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9923122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Selection of Antarctic yeasts as gray mold biocontrol agents in strawberry.","authors":"Eskálath Morganna Silva Ferreira, Gabriela Garmendia, Vívian Nicolau Gonçalves, Juliana Fonseca Moreira da Silva, Luiz Henrique Rosa, Silvana Vero, Raphael Sanzio Pimenta","doi":"10.1007/s00792-023-01298-z","DOIUrl":"https://doi.org/10.1007/s00792-023-01298-z","url":null,"abstract":"<p><p>The postharvest disease popularly known as gray mold is considered one of the most limiting factors strawberry fruit production. The most effective way to control this disease is still the use of chemical fungicides. However, other alternative sources of control are being explored. Among these, psychrophilic yeasts adapted to extreme conditions, such as those found in the Antarctic region, may have great potential for use as biocontrol agents. Thus, the present study aimed to select psychrotolerant yeasts obtained from Antarctic region and to evaluate their potential for biocontrol under gray mold, caused by Botrytis cinerea in strawberries stored at low temperature. For this, 20 potential antagonist yeasts were evaluated in vitro (thermotolerance and enzymatic) assays. Debaryomyces hansenii, Rhodotorula mucilaginosa and Dioszegia hungarica were selected for growing in strawberry juice. However, only D. hansenii was selected for in vivo studies and showed a reduction in the incidence of gray mold by 82% for the tests performed on injury and 86% for the tests on non-injured fruits treated by immersion bath. Thus, demonstrating that the selection of this cold-adapted Antarctic yeast can be a promising strategy as a biocontrol agent used to curb the development of gray mold in strawberry fruits.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"16"},"PeriodicalIF":2.9,"publicationDate":"2023-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9871793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-07-03DOI: 10.1007/s00792-023-01303-5
Xin-Xin Li, Shun Tan, Mu Cheng, Yao Hu, Xue Ma, Jing Hou, Heng-Lin Cui
{"title":"Salinilacihabitans rarus gen. nov., sp. nov., Natrononativus amylolyticus gen. nov., sp. nov., Natronobeatus ordinarius gen. nov., sp. nov., and Halovivax gelatinilyticus sp. nov., halophilic archaea, isolated from a salt lake and soda lakes.","authors":"Xin-Xin Li, Shun Tan, Mu Cheng, Yao Hu, Xue Ma, Jing Hou, Heng-Lin Cui","doi":"10.1007/s00792-023-01303-5","DOIUrl":"https://doi.org/10.1007/s00792-023-01303-5","url":null,"abstract":"<p><p>Four halophilic archaea strains, AD-4<sup>T</sup>, CGA30<sup>T</sup>, CGA73<sup>T</sup>, and WLHSJ27<sup>T</sup>, were isolated from a salt lake and two soda lakes located in different regions of China. The 16S rRNA and rpoB' gene sequence similarities among strains AD-4<sup>T</sup>, CGA30<sup>T</sup>, CGA73<sup>T</sup>, WLHSJ27<sup>T</sup>, and the current species of the family Natrialbaceae were 90.9-97.5% and 83.1-91.8%, respectively. The phylogenetic and phylogenomic analyses revealed that these four strains separated from existing genera in the family Natrialbaceae and formed distant branches. The ANI, isDDH, and AAI values among these four strains and the current members of the family Natrialbaceae were 72-79%, 20-25%, and 63-73%, respectively, much lower than the threshold values for species demarcation. Strains AD-4<sup>T</sup>, CGA73<sup>T</sup>, and WLHSJ27<sup>T</sup> may represent three novel genera of the family Natrialbaceae according to the cutoff value of AAI (≤ 76%) proposed to differentiate genera within the family Natrialbaceae. These four strains could be distinguished from the related genera according to differential phenotypic characteristics. The major phospholipids of these four strains were identical while their glycolipid profiles were diverse. DGD-1 is a major glycolipid found in strain AD-4<sup>T</sup>, trace glycolipids, DGD-1, and S-DGD-1, and (or) S-TGD-1 was found in the other three strains. The major respiratory quinones detected in the four strains were menaquinone MK-8 and MK-8(H<sub>2</sub>). This polyphasic classification indicated that strains AD-4<sup>T</sup>, CGA73<sup>T</sup>, and WLHSJ27<sup>T</sup> represent three novel species of three new genera with the family Natrialbaceae, and strain CGA30<sup>T</sup> represents a novel species of Halovivax.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"15"},"PeriodicalIF":2.9,"publicationDate":"2023-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9753691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-06-24DOI: 10.1007/s00792-023-01301-7
Hajra Kayani, Muhammad Asif Rasheed, Wadi B Alonazi, Farrukh Jamil, Annam Hussain, ChangHui Yan, Raza Ahmed, Muhammad Ibrahim
{"title":"Identification and genome-wide analysis provide insights into the genetic diversity and biotechnological potentials of novel cold-adapted Acinetobacter strain.","authors":"Hajra Kayani, Muhammad Asif Rasheed, Wadi B Alonazi, Farrukh Jamil, Annam Hussain, ChangHui Yan, Raza Ahmed, Muhammad Ibrahim","doi":"10.1007/s00792-023-01301-7","DOIUrl":"https://doi.org/10.1007/s00792-023-01301-7","url":null,"abstract":"<p><p>Extreme cold environments, such as polar regions or high-altitude mountains, are known for their challenging conditions including low temperatures, high salinity, and limited nutrient availability. Microbes that thrive in these environments have evolved specialized strategies to survive and function under such harsh conditions. The study aims to identify, sequence the genome, perform genome assembly, and conduct a comparative genome-wide analysis of Acinetobacter sp. strain P1, which was isolated from the Batura glacier regions of Pakistan. A basic local alignment search tool of NCBI using 16 s RNA gene sequence confirmed the strain Acinetobacter following phylogenetic analysis revealed that strain P1 clustered with Acinetobacter sp. strain AcBz01. The high-throughput Genome sequencing was done by the NovaSeq 6000 sequencing system following de novo genome assembly reported 23 contigs, a genome size of 3,732,502 bp containing approximately 3489 genes and 63 RNAs (60 tRNA, 3 rRNA). The comparative genome analysis revealed that Acinetobacter sp. strain P1 exhibited the highest homology with the Acinetobacter baumannii ATCC 17978 genome and encompassed 1668 indispensable genes, 1280 conserved genes 1821 specific genes suggesting high genomic plasticity and evolutionary diversity. The genes with functional assignments include exopolysaccharide phosphotransferase enzyme, cold-shock proteins, T6SS, membrane modifications, antibiotic resistance, and set of genes related to a wide range of metabolic characteristics such as exopolysaccharides were also present. Moreover, the structural prediction analysis of EPS proteins reveals that structural flexibility allows for conformational modifications during catalysis, which boosts or increases the catalytic effectiveness at lower temperatures. Overall, the identification of Acinetobacter, a cold-adapted bacterium, offers promising applications in bioremediation, enzyme production, food preservation, pharmaceutical development, and astrobiology. Further research and exploration of these microorganisms can unlock their full biotechnological potential and contribute to various industries and scientific endeavors.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"14"},"PeriodicalIF":2.9,"publicationDate":"2023-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9686132","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-06-22DOI: 10.1007/s00792-023-01300-8
Seema Prabhudev Rodge, Kundalik Shivaji Shende, Niranjan Prakashrao Patil
{"title":"Polyhydroxyalkanoate biosynthesis and optimisation of thermophilic Geobacillus stearothermophilus strain K4E3_SPR_NPP.","authors":"Seema Prabhudev Rodge, Kundalik Shivaji Shende, Niranjan Prakashrao Patil","doi":"10.1007/s00792-023-01300-8","DOIUrl":"https://doi.org/10.1007/s00792-023-01300-8","url":null,"abstract":"<p><p>Polyhydroxyalkanoates (PHA) can be used to combat the challenges associated with plastic because it is biodegradable and can be produced from renewable resources. Extremophiles are considered to be potential PHA producers. An initial screening for the PHA synthesizing ability of a thermophilic bacteria Geobacillus stearothermophilus strain K4E3_SPR_NPP was carried out using Sudan black B staining. Nile red viable colony staining was used to further verify that the isolates produced PHA. Crotonic acid assays were used to determine the concentrations of PHA. The bacteria showed 31% PHA accumulation per dry cell weight (PHA/DCW) when glucose was used as a carbon source for growth. The molecule was identified to be medium chain length PHA, A copolymer of PHA containing poly(3-hydroxybutyrate)-poly(3-hydroxyvalerate)-poly(3-hydroxyhexanoate) (PHB-PHV-PHH<sub>X</sub>) using 1H-NMR. Six carbon sources and four nitrogen sources were screened for the synthesis of maximum PHA content, of which lactose and ammonium nitrate showed 45% and 53% PHA/DCW respectively. The important factors in the experiment are identified using the Plackett-Burman design, and optimization is performed using the response surface method. Response surface methodology was used to optimize the three important factors, and the maximum biomass and PHA productions were discovered. Optimal concentrations yielded a maximum of 0.48 g/l biomass and 0.32 g/l PHA, measuring 66.66% PHA accumulation. Dairy industry effluent was employed for the synthesis of PHA, yielding 0.73 g/l biomass and 0.33 g/l PHA, measuring 45% PHA accumulation. These findings add credibility to the possibility of adopting thermophilic isolates for PHA production using low-cost substrates.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"13"},"PeriodicalIF":2.9,"publicationDate":"2023-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9738986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-05-13DOI: 10.1007/s00792-023-01297-0
M A Khomyakova, A Yu Merkel, V S Segliuk, A I Slobodkin
{"title":"Desulfatitalea alkaliphila sp. nov., an alkalipilic sulfate- and arsenate- reducing bacterium isolated from a terrestrial mud volcano.","authors":"M A Khomyakova, A Yu Merkel, V S Segliuk, A I Slobodkin","doi":"10.1007/s00792-023-01297-0","DOIUrl":"https://doi.org/10.1007/s00792-023-01297-0","url":null,"abstract":"<p><p>A novel alkaliphilic sulfate-reducing bacterium, strain M08but<sup>T</sup>, was isolated from a salsa lake of terrestrial mud volcano (Taman Peninsula, Russia). Cells were rod-shaped, motile and Gram-stain-negative. The temperature range for growth was 15-42 °C (optimum at 30 °C). The pH range for growth was 7.0-11.0, with an optimum at pH 8.5-9.0 Strain M08but<sup>T</sup> used sulfate, thiosulfate, sulfite, dimethyl sulfoxide and arsenate as electron acceptors. Acetate, formate, butyrate, fumarate, succinate, glycerol and pyruvate were utilized as electron donors with sulfate. Fermentative growth was observed with fumarate, pyruvate, crotonate. Strain M08but<sup>T</sup> grew chemolithoautotrophically with H<sub>2</sub> and CO<sub>2</sub>. The G + C content of the genomic DNA was 60.1%. The fatty acid profile of strain M08but<sup>T</sup> was characterized by the presence of anteiso-C<sub>15:0</sub> as the major component (68.8%). The closest phylogenetic relative of strain M08but<sup>T</sup> was Desulfatitalea tepidiphila (the order Desulfobacterales) with 96.3% 16S rRNA gene sequence similarity. Based on the phenotypic, genotypic and phylogenetic characteristics of the isolate, strain M08but<sup>T</sup> is considered to represent a novel species of the genus Desulfatitalea, with proposed name Desulfatitalea alkaliphila sp. nov. The type strain of Desulfatitalea alkaliphila is M08but<sup>T</sup> (= KCTC 25382<sup>T</sup> = VKM B-3560<sup>T</sup> = DSM 113909<sup>T</sup> = JCM 39202<sup>T</sup> = UQM 41473<sup>T</sup>).</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"12"},"PeriodicalIF":2.9,"publicationDate":"2023-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9516473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ExtremophilesPub Date : 2023-05-13DOI: 10.1007/s00792-023-01296-1
Wenhao Hu, Jin Wang, Juan Li, Mengxia Yang, Zhixing Li, Xuning Zhang, Fang Wu, Yaqi Zhang, Zhidan Luo, Henghao Xu
{"title":"Improvement of duplex-specific nuclease salt tolerance by fusing DNA-binding domain of DNase from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix.","authors":"Wenhao Hu, Jin Wang, Juan Li, Mengxia Yang, Zhixing Li, Xuning Zhang, Fang Wu, Yaqi Zhang, Zhidan Luo, Henghao Xu","doi":"10.1007/s00792-023-01296-1","DOIUrl":"https://doi.org/10.1007/s00792-023-01296-1","url":null,"abstract":"<p><p>Salt tolerance is an important property of duplex-specific nuclease (DSN). DSN with high salt tolerance can be more widely used in genetic engineering, especially in the production of nucleic acid drugs. To improve the salt tolerance of DSN, we selected five DNA-binding domains from extremophilic organisms, which have been shown the ability to improve salt tolerance of DNA polymerases and nucleases. The experimental results demonstrated that the fusion protein TK-DSN produced by fusing a N-terminal DNA-binding domain, which comprised two HhH (helix-hairpin-helix) motifs domain from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix, has a significantly improved salt tolerance. TK-DSN can tolerate the concentration of NaCl up to 800 mM; in addition, the ability of digesting DNA was also enhanced during in vitro transcription and RNA purification. This strategy provides the method for the personalized customization of biological tool enzymes for different applications.</p>","PeriodicalId":12302,"journal":{"name":"Extremophiles","volume":"27 2","pages":"11"},"PeriodicalIF":2.9,"publicationDate":"2023-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9464144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}