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Lipase metagenomic

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https://www.readbyqxmd.com/read/28887679/microbiological-studies-of-hot-springs-in-india-a-review
#1
REVIEW
Abhijit Poddar, Subrata K Das
The earliest microbiological studies on hot springs in India date from 2003, a much later date compared to global attention in this striking field of study. As of today, 28 out of 400 geothermal springs have been explored following both culturable and non-culturable approaches. The temperatures and pH of the springs are 37-99 °C and 6.8-10, respectively. Several studies have been performed on the description of novel genera and species, characterization of different bio-resources, metagenomics of hot spring microbiome and whole genome analysis of few isolates...
September 8, 2017: Archives of Microbiology
https://www.readbyqxmd.com/read/28532384/in-silico-approach-to-designing-rational-metagenomic-libraries-for-functional-studies
#2
Anna Kusnezowa, Lars I Leichert
BACKGROUND: With the development of Next Generation Sequencing technologies, the number of predicted proteins from entire (meta-) genomes has risen exponentially. While for some of these sequences protein functions can be inferred from homology, an experimental characterization is still a requirement for the determination of protein function. However, functional characterization of proteins cannot keep pace with our capabilities to generate more and more sequence data. RESULTS: Here, we present an approach to reduce the number of proteins from entire (meta-) genomes to a reasonably small number for further experimental characterization without loss of important information...
May 22, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28523377/mining-lipolytic-enzymes-in-community-dna-from-high-andean-soils-using-a-targeted-approach
#3
Daniel Borda-Molina, José Salvador Montaña, María Mercedes Zambrano, Sandra Baena
Microbial enrichments cultures are a useful strategy to speed up the search for enzymes that can be employed in industrial processes. Lipases have gained special attention because they show unique properties such as: broad substrate specificity, enantio- and regio-selectivity and stability in organic solvents. A major goal is to identify novel lipolytic enzymes from microorganisms living in cold extreme environments such as high Andean soils, of relevance to our study being their capability be used in industrial processes...
August 2017: Antonie Van Leeuwenhoek
https://www.readbyqxmd.com/read/28459005/discovery-and-characterizaton-of-a-novel-lipase-with-transesterification-activity-from-hot-spring-metagenomic-library
#4
Wei Yan, Furong Li, Li Wang, Yaxin Zhu, Zhiyang Dong, Linhan Bai
A new gene encoding a lipase (designated as Lip-1) was identified from a metagenomic bacterial artificial chromosome(BAC) library prepared from a concentrated water sample collected from a hot spring field in Niujie, Eryuan of Yunnan province in China. The open reading frame of this gene encoded 622 amino acid residues. It was cloned, fused with the oleosin gene and over expressed in Escherichia coli to prepare immobilized lipase artificial oil body AOB-sole-lip-1. The monomeric Sole-lip-1 fusion protein presented a molecular mass of 102...
March 2017: Biotechnology Reports
https://www.readbyqxmd.com/read/28457984/screening-and-characterization-of-a-novel-thermostable-lipase-with-detergent-additive-potential-from-the-metagenomic-library-of-a-mangrove-soil
#5
Leli Tang, Yulin Xia, Xiuling Wu, Xiaoli Chen, Xueling Zhang, He Li
One clone (Lip906) exhibiting lipase activity was screened from a metagenomic library by using a medium containing tricaprylin. A novel lipase gene from the inserted fragment of Lip906 was obtained by sequencing. The phylogenetic analysis of Lip906 lipase exhibited 34% and 32% homologue to lipases from Streptomyces sp. MspMP-M5 and Rhodopirellula europaea. This gene was expressed in Escherichia coli (E. coli) BL21 (DE3), and the recombinant protein was purified and characterized. The best substrate of the recombinant Lip906 lipase was p-nitrophenyl myristate (C14)...
August 20, 2017: Gene
https://www.readbyqxmd.com/read/28331945/from-a-metagenomic-source-to-a-high-resolution-structure-of-a-novel-alkaline-esterase
#6
Mariana Rangel Pereira, Thaís Carvalho Maester, Gustavo Fernando Mercaldi, Eliana Gertrudes de Macedo Lemos, Marko Hyvönen, Andrea Balan
Esterases catalyze the cleavage and formation of ester bonds and are members of the diverse family of α/β hydrolase fold. They are useful in industries from different sectors, such as food, detergent, fine chemicals, and biofuel production. In a previous work, 30 positive clones for lipolytic activity were identified from a metagenomic library of a microbial consortium specialized in diesel oil degradation. In this study, a putative gene encoding an esterase/lipase, denominated est8, has been cloned and the corresponding protein expressed recombinantly, purified to homogeneity and characterized functional and structurally...
March 22, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28272521/activity-screening-of-environmental-metagenomic-libraries-reveals-novel-carboxylesterase-families
#7
Ana Popovic, Tran Hai, Anatoly Tchigvintsev, Mahbod Hajighasemi, Boguslaw Nocek, Anna N Khusnutdinova, Greg Brown, Julia Glinos, Robert Flick, Tatiana Skarina, Tatyana N Chernikova, Veronica Yim, Thomas Brüls, Denis Le Paslier, Michail M Yakimov, Andrzej Joachimiak, Manuel Ferrer, Olga V Golyshina, Alexei Savchenko, Peter N Golyshin, Alexander F Yakunin
Metagenomics has made accessible an enormous reserve of global biochemical diversity. To tap into this vast resource of novel enzymes, we have screened over one million clones from metagenome DNA libraries derived from sixteen different environments for carboxylesterase activity and identified 714 positive hits. We have validated the esterase activity of 80 selected genes, which belong to 17 different protein families including unknown and cyclase-like proteins. Three metagenomic enzymes exhibited lipase activity, and seven proteins showed polyester depolymerization activity against polylactic acid and polycaprolactone...
March 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28044227/a-novel-versatile-family-iv-carboxylesterase-exhibits-high-stability-and-activity-in-a-broad-ph-spectrum
#8
Amélie Dukunde, Dominik Schneider, Mingji Lu, Silja Brady, Rolf Daniel
OBJECTIVES: To investigate the properties of a novel metagenome-derived member of the hormone-sensitive lipase family of lipolytic enzymes. RESULTS: A forest soil metagenome-derived gene encoding an esterase (Est06) belonging to the hormone-sensitive lipase family of lipolytic enzymes was subcloned, heterologously expressed and characterized. Est06 is a polypeptide of 295 amino acids with a molecular mass of 31 kDa. The deduced protein sequence shares 61% similarity with a hypothetical protein from the marine symbiont Candidatus Entotheonella sp...
April 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/27984150/enantiomeric-separation-of-pharmaceutically-important-drug-intermediates-using-a-metagenomic-lipase-and-optimization-of-its-large-scale-production
#9
Rakesh Kumar, Linga Banoth, Uttam Chand Banerjee, Jagdeep Kaur
In the present study, efficient enzymatic methods were developed using a recombinant metagenomic lipase (LipR1) for the synthesis of corresponding esters by the transesterification of five different pharmaceutically important secondary alcohols. The recombinant lipase (specific activity=87m6U/mg) showed maximum conversion in presence of ionic liquid with Naphthyl-ethanol (eeP=99%), Indanol and Methyl-4 pyridine methanol (eeS of 98% and 99%) respectively in 1h. Vinyl acetate was found as suitable acyl donor in transesterification reactions...
February 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/27896615/bioprospecting-hot-spring-metagenome-lipase-for-the-production-of-biodiesel
#10
Rajesh Kumar Sahoo, Mohit Kumar, Lala Behari Sukla, Enketeswara Subudhi
Screening of metagenomic library from Taptapani Hot Spring (Odisha) yielded a positive lipase clone (pUC-lip479). Sequence analysis showed an ORF (RK-lip479) of 416 amino acid residues which was overexpressed in Escherichia coli BL21 (DE3). Optimum pH and temperature of purified lipase RK-lip479 were 8.0 and 65 °C, respectively, and found to be stable over a pH range of 7.0-9.0 and temperatures 55-75 °C. RK-lip479 could hydrolyse a wide range of 4-nitrophenyl esters (4-nitrophenyoctanoate, 4-nitrophenyldodecanoate, 4-nitrophenylpalmitate, 4-nitrophenylmyristate and 4-nitrophenylstearate), and maximum activity was observed with 4-nitrophenyldodecanoate...
February 2017: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/27770744/computational-modelling-and-protein-ligand-interaction-studies-of-smlipa-lipase-cloned-from-forest-metagenome
#11
Mahejibin Khan, Amit Kumar
The understanding of the 3-dimensional enzyme structure is important for the point of protein engineering and applications. Computer-based molecular modelling is a vital tool for theoretical predication of enzyme activities and finding their substrates and inhibitors. SMlipA lipase was cloned from forest soil metagenome and characterized as broad spectrum enzyme with high stability in various organic solvents. In the present study, to understand the mechanism of SMlipA lipase and to identify the key residues involved in enzyme-substrate interaction, three dimensional-computational model of SMlipA has been generated and validated for stereo-chemical and amino-acid environment quality using appropriate programs, and further validation of the active-site architecture was achieved by performing docking studies with different ligand...
November 2016: Journal of Molecular Graphics & Modelling
https://www.readbyqxmd.com/read/27753534/new-member-of-the-hormone-sensitive-lipase-family-from-the-permafrost-microbial-community
#12
Lada E Petrovskaya, Ksenia A Novototskaya-Vlasova, Sultan Sh Gapizov, Elena V Spirina, Ekaterina V Durdenko, Elizaveta M Rivkina
Siberian permafrost is a unique environment inhabited with diverse groups of microorganisms. Among them, there are numerous producers of biotechnologically relevant enzymes including lipases and esterases. Recently, we have constructed a metagenomic library from a permafrost sample and identified in it several genes coding for potential lipolytic enzymes. In the current work, properties of the recombinant esterases obtained from this library are compared with the previously characterized lipase from Psychrobacter cryohalolentis and other representatives of the hormone-sensitive lipase family...
July 4, 2017: Bioengineered
https://www.readbyqxmd.com/read/27584951/multi-enzyme-screening-using-a-high-throughput-genetic-enzyme-screening-system
#13
Haseong Kim, Kil Koang Kwon, Wonjae Seong, Seung-Goo Lee
The recent development of a high-throughput single-cell assay technique enables the screening of novel enzymes based on functional activities from a large-scale metagenomic library(1). We previously proposed a genetic enzyme screening system (GESS) that uses dimethylphenol regulator activated by phenol or p-nitrophenol. Since a vast amount of natural enzymatic reactions produce these phenolic compounds from phenol deriving substrates, this single genetic screening system can be theoretically applied to screen over 200 different enzymes in the BRENDA database...
August 8, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27328716/structural-insights-of-a-hormone-sensitive-lipase-homologue-est22
#14
Jing Huang, Ying-Yi Huo, Rui Ji, Siyun Kuang, Chaoneng Ji, Xue-Wei Xu, Jixi Li
Hormone sensitive lipase (HSL) catalyzes the hydrolysis of triacylglycerols into fatty acids and glycerol, thus playing key roles in energy homeostasis. However, the application of HSL serving as a pharmaceutical target and an industrial biocatalyst is largely hampered due to the lack of high-resolution structural information. Here we report biochemical properties and crystal structures of a novel HSL homologue esterase Est22 from a deep-sea metagenomic library. Est22 prefers short acyl chain esters and has a very high activity with substrate p-nitrophenyl butyrate...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27271534/metagenomic-discovery-of-novel-enzymes-and-biosurfactants-in-a-slaughterhouse-biofilm-microbial-community
#15
Stephan Thies, Sonja Christina Rausch, Filip Kovacic, Alexandra Schmidt-Thaler, Susanne Wilhelm, Frank Rosenau, Rolf Daniel, Wolfgang Streit, Jörg Pietruszka, Karl-Erich Jaeger
DNA derived from environmental samples is a rich source of novel bioactive molecules. The choice of the habitat to be sampled predefines the properties of the biomolecules to be discovered due to the physiological adaptation of the microbial community to the prevailing environmental conditions. We have constructed a metagenomic library in Escherichia coli DH10b with environmental DNA (eDNA) isolated from the microbial community of a slaughterhouse drain biofilm consisting mainly of species from the family Flavobacteriaceae...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27263017/integrative-computational-approach-for-genome-based-study-of-microbial-lipid-degrading-enzymes
#16
REVIEW
Tayvich Vorapreeda, Chinae Thammarongtham, Kobkul Laoteng
Lipid-degrading or lipolytic enzymes have gained enormous attention in academic and industrial sectors. Several efforts are underway to discover new lipase enzymes from a variety of microorganisms with particular catalytic properties to be used for extensive applications. In addition, various tools and strategies have been implemented to unravel the functional relevance of the versatile lipid-degrading enzymes for special purposes. This review highlights the study of microbial lipid-degrading enzymes through an integrative computational approach...
July 2016: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/27083848/point-mutation-gln121-arg-increased-temperature-optima-of-bacillus-lipase-1-4-subfamily-by-fifteen-degrees
#17
Shelly Goomber, Rakesh Kumar, Ranvir Singh, Neelima Mishra, Jagdeep Kaur
Small molecular weight Bacillus lipases are industrially attractive because of its alkaline optimum pH, broad substrate specificity and production in high yield by overexpression both in Escherichia coli and Bacillus subtilis. Its major limitation of being mesophilic in nature is constantly targeted by laboratory evolution studies. Herein metagenomically isolated Bacillus LipJ was randomly evolved by error prone PCR and library of variants were screened for enhanced thermostability. Point mutant Gln121Arg was extensively characterized and it showed dramatic shift of Temp...
July 2016: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/26929439/expression-and-characterization-of-a-new-esterase-with-gcsag-motif-from-a-permafrost-metagenomic-library
#18
Lada E Petrovskaya, Ksenia A Novototskaya-Vlasova, Elena V Spirina, Ekaterina V Durdenko, Galina Yu Lomakina, Maria G Zavialova, Evgeny N Nikolaev, Elizaveta M Rivkina
As a result of construction and screening of a metagenomic library prepared from a permafrost-derived microcosm, we have isolated a novel gene coding for a putative lipolytic enzyme that belongs to the hormone-sensitive lipase family. It encodes a polypeptide of 343 amino acid residues whose amino acid sequence displays maximum likelihood with uncharacterized proteins from Sphingomonas species. A putative catalytic serine residue of PMGL2 resides in a new variant of a recently discovered GTSAG sequence in which a Thr residue is replaced by a Cys residue (GCSAG)...
May 2016: FEMS Microbiology Ecology
https://www.readbyqxmd.com/read/26915995/characterization-of-est3-a-metagenome-derived-esterase-with-suitable-properties-for-biotechnological-applications
#19
Thaís Carvalho Maester, Mariana Rangel Pereira, E G Machado Sierra, Andrea Balan, Eliana Gertrudes de Macedo Lemos
Metagenomic libraries from diverse environments have been extensive sources of many lipases and esterases; nevertheless, most of these enzymes remain biochemically uncharacterized. We previously built a metagenomic fosmid library from a microbial consortium specialized for diesel oil degradation and tested it for lipolytic activity. In the present study, we identified the PL14.H10 clone that was subcloned and sequenced, which enabled the identification of the EST3 protein. This enzyme exhibited 74 % amino acid identity with the uncharacterized alpha/beta hydrolase from Parvibaculum lavamentivorans [GenBank: WP012110575...
July 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/26782782/characterization-of-a-cold-active-and-salt-tolerant-esterase-identified-by-functional-screening-of-arctic-metagenomic-libraries
#20
Concetta De Santi, Bjørn Altermark, Marcin Miroslaw Pierechod, Luca Ambrosino, Donatella de Pascale, Nils-Peder Willassen
BACKGROUND: The use of metagenomics in enzyme discovery constitutes a powerful approach to access to genomes of unculturable community of microorganisms and isolate novel valuable biocatalysts for use in a wide range of biotechnological and pharmaceutical fields. RESULTS: Here we present a novel esterase gene (lip3) identified by functional screening of three fosmid metagenomic libraries, constructed from three marine sediment samples. The sequenced positive fosmid revealed an enzyme of 281 amino acids with similarity to class 3 lipases...
January 19, 2016: BMC Biochemistry
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