Screening and Expression of Genes from Metagenomes

Leis, Benedikt; Angelov, Angel; Liebl, Wolfgang

doi:10.1016/b978-0-12-407678-5.00001-5

Cited by 49 publications

(39 citation statements)

References 238 publications

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“…The metagenome contains a large number of functional genes. Therefore, metagenomic analyses facilitate the discovery of novel genes in biological communities by overcoming the limitations of cultivation-dependent approaches (Guazzaroni et al 2015;Leis et al 2013). Kellner et al analyzed the diversity of bacterial laccase genes in samples from forests and grasslands and found 16 distinct sequence-type clades of bacterial laccase (Kellner et al 2008).…”

Section: Open Accessmentioning

confidence: 99%

Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge

Yue

Yang

Zhao

et al. 2017

Bioresour. Bioprocess.

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Background:The metagenome contains plenty of genetic resources and can be used to search for the novel gene or mutant. Results:In this study, the bacterial laccase gene (cueO) with single or multiple mutations was directly cloned based on the metagenome of a chemical plant sludge. An interesting mutation (G276R) was identified from those cloned mutants. The other mutants (G276N, G276Y, and G276K) with improved catalytic efficiency were identified by the saturation mutagenesis on residue G276. The optimal temperature for wild-type CueO enzyme activity was about 70 °C, compared to 60 °C, 50 °C, 50 °C, and 30 °C for the G276R, G276N, G276Y, and G276K mutant enzymes, respectively. The catalytic efficiency (k cat /K m ) with 8 mmol Cu 2+ of the G276R, G276N, G276Y, and G276K mutants was 1.2-, 2.7-, 1.3-, and 2.7-fold, respectively, compared to the wild-type enzyme. In addition, the mutants G276R, G276N, G276Y, and G276K oxidized the carcinogen benzo[α]pyrene more efficiently compared to the wild-type enzyme. Conclusion:All of the results indicate that G276 of CueO plays an important role in enzyme activity, and the useful mutants can be identified based on the metagenome.

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Section: Open Accessmentioning

confidence: 99%

Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge

Yue

Yang

Zhao

et al. 2017

Bioresour. Bioprocess.

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“…Bias in sampling may be introduced by any kind of treatment of an environmental sample prior to sequencing (Leis et al, 2013). In some cases, bias is desired, as discussed in Section 3.2, to enrich for desirable microorganisms and limit the metagenomic diversity to be analyzed.…”

Section: Capturing Diversitymentioning

confidence: 99%

“…A recent review (Leis, Angelov, & Liebl, 2013) has addressed these issues in some detail so they are discussed only briefly here.…”

Section: Introductionmentioning

confidence: 99%

Bioprospecting in the Genomic Age

Hicks¹,

Prather²

2014

Advances in Applied Microbiology

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“…This approach has been extensively employed for mining new biocatalysts to improve many industrially relevant processes in biotechnology and pharmaceutics (Leis et al 2013). Several metagenome-derived cellulase genes have been identified in metagenomic libraries made from natural microbial communities such as cow rumen, termite gut, soil, and plant-or algae-associated microorganisms Bhat et al 2013;Duan et al 2009;Martin et al 2014;Ni and Tokuda 2013;Voget et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Improved ethanol production from biomass by a rumen metagenomic DNA fragment expressed in Escherichia coli MS04 during fermentation

Loaces

Amarelle

Muñoz-Gutiérrez

et al. 2015

Appl Microbiol Biotechnol

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With the aim of improving current ethanologenic Escherichia coli strains, we screened a metagenomic library from bovine ruminal fluid for cellulolytic enzymes. We isolated one fosmid, termed Csd4, which was able to confer to E. coli the ability to grow on complex cellulosic material as the sole carbon source such as avicel, carboxymethyl cellulose, filter paper, pretreated sugarcane bagasse, and xylan. Glucanolytic activity obtained from E. coli transformed with Csd4 was maximal at 24 h of incubation and was inhibited when glucose or xylose were present in the media. The 34,406-bp DNA fragment of Csd4 was completely sequenced, and a putative endoglucanase, a xylosidase/arabinosidase, and a laccase gene were identified. Comparison analysis revealed that Csd4 derived from an organism closely related to Prevotella ruminicola, but no homologies were found with any of the genomes already sequenced. Csd4 was introduced into the ethanologenic E. coli MS04 strain and ethanol production from CMC, avicel, sugarcane bagasse, or filter paper was observed. Exogenously expressed β-glucosidase had a positie effect on cell growth in agreement with the fact that no putative β-glucosidase was found in Csd4. Ethanol production from sugarcane bagasse was improved threefold by Csd4 after saccharification by commercial Trichoderma reesei cellulases underlining the ability of Csd4 to act as a saccharification enhancer to reduce the enzymatic load and time required for cellulose deconstruction.

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Screening and Expression of Genes from Metagenomes

Cited by 49 publications

References 238 publications

Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge

Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge

Bioprospecting in the Genomic Age

Improved ethanol production from biomass by a rumen metagenomic DNA fragment expressed in Escherichia coli MS04 during fermentation

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