Cellulases from Thermophiles Found by Metagenomics

Escuder-Rodríguez, Juan-José; DeCastro, María-Eugenia; Cerdán, M. Esperanza; Rodrı́guez-Belmonte, Esther; Becerra, Manuel; González-Siso, María-Isabel

doi:10.3390/microorganisms6030066

Cited by 50 publications

(32 citation statements)

References 177 publications

(138 reference statements)

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“…Cellulases are a group of enzymes catalyzing cellulose polymers into simpler sugars [59]. This enzyme is useful for the paper industry, cotton processing, and detergents [60].…”

Section: Cellulasesmentioning

confidence: 99%

Metagenomic applications in exploration and development of novel enzymes from nature: a review

Prayogo

Budiharjo

Kusumaningrum

et al. 2020

Journal of Genetic Engineering and Biotechnology

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Background: Microbial community has an essential role in various fields, especially the industrial sector. Microbes produce metabolites in the form of enzymes, which are one of the essential compounds for industrial processes. Unfortunately, there are still numerous microbes that cannot be identified and cultivated because of the limitations of the culture-based method. The metagenomic approach is a solution for researchers to overcome these problems. The main body of the abstract: Metagenomics is a strategy used to analyze the genomes of microbial communities in the environment directly. Metagenomics application used to explore novel enzymes is essential because it allows researchers to obtain data on microbial diversity, reaching of 99% and various types of genes encoding an enzyme that has not yet been identified. Basic methods in metagenomics have been developed and are commonly used in various studies. A basic understanding of metagenomics for researchers is needed, especially young researchers to support the success of the research. Short conclusion: Therefore, this review was done in order to provide a deep understanding of metagenomics. It also discussed the application and basic methods of metagenomics in the exploration of novel enzymes, especially in the latest research. Several types of enzymes, such as cellulases, proteases, and lipases, which have been explored using metagenomics, were reviewed in this article.

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“…Cellulases are a group of enzymes catalyzing cellulose polymers into simpler sugars [59]. This enzyme is useful for the paper industry, cotton processing, and detergents [60].…”

Section: Cellulasesmentioning

confidence: 99%

Metagenomic applications in exploration and development of novel enzymes from nature: a review

Prayogo

Budiharjo

Kusumaningrum

et al. 2020

Journal of Genetic Engineering and Biotechnology

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“…Xylanases are of interest in the food and feed industries, paper and pulp technology, as well as in textile and biofuel production 2 . Hot springs are an interesting source of novel enzymes with potential biotechnological applications because extremely hot environments are inhabited by specialized microorganisms adapted to high temperatures and extreme pHs 4 . Many hot spring-derived xylanases have been recovered from cultured microorganisms 5,6 .…”

Section: Introductionmentioning

confidence: 99%

Microbial diversity analysis and screening for novel xylanase enzymes from the sediment of the Lobios Hot Spring in Spain

Knapik

Becerra

González-Siso

2019

Sci Rep

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Here, we describe the metagenome composition of a microbial community in a hot spring sediment as well as a sequence-based and function-based screening of the metagenome for identification of novel xylanases. The sediment was collected from the Lobios Hot Spring located in the province of Ourense (Spain). Environmental DNA was extracted and sequenced using Illumina technology, and a total of 3.6 Gbp of clean paired reads was produced. A taxonomic classification that was obtained by comparison to the NCBI protein nr database revealed a dominance of Bacteria (93%), followed by Archaea (6%). The most abundant bacterial phylum was Acidobacteria (25%), while Thaumarchaeota (5%) was the main archaeal phylum. Reads were assembled into contigs. Open reading frames (ORFs) predicted on these contigs were searched by BLAST against the CAZy database to retrieve xylanase encoding ORFs. A metagenomic fosmid library of approximately 150,000 clones was constructed to identify functional genes encoding thermostable xylanase enzymes. Function-based screening revealed a novel xylanase-encoding gene ( XynA3 ), which was successfully expressed in E . coli BL21. The resulting protein (41 kDa), a member of glycoside hydrolase family 11 was purified and biochemically characterized. The highest activity was measured at 80 °C and pH 6.5. The protein was extremely thermostable and showed 94% remaining activity after incubation at 60 °C for 24 h and over 70% remaining activity after incubation at 70 °C for 24 h. Xylanolytic activity of the XynA3 enzyme was stimulated in the presence of β-mercaptoethanol, dithiothreitol and Fe 3+ ions. HPLC analysis showed that XynA3 hydrolyzes xylan forming xylobiose with lower proportion of xylotriose and xylose. Specific activity of the enzyme was 9080 U/mg for oat arabinoxylan and 5080 U/mg for beechwood xylan, respectively, without cellulase activity.

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“…The temperature optima of cellulases in currently available commercial enzyme cocktails are typically around 50°C [7], i.e., not particularly high and likely not optimal, for example considering the risk of microbial contamination. The introduction of thermostable enzymes could be beneficial since this would allow the use of higher temperatures, resulting in increased substrate solubility, lower viscosity and reduced microbial growth [8]. Furthermore, the use of thermostable cellulases can simplify process design by minimizing or eliminating cooling periods between stages that require different temperatures (e.g.…”

Section: Introductionmentioning

confidence: 99%

Identification and characterization of a hyperthermophilic GH9 cellulase from the Arctic Mid-Ocean Ridge vent field

et al. 2019

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A novel GH9 cellulase (AMOR_GH9A) was discovered by sequence-based mining of a unique metagenomic dataset collected at the Jan Mayen hydrothermal vent field. AMOR_GH9A comprises a signal peptide, a catalytic domain and a CBM3 cellulose-binding module. AMOR_GH9A is an exceptionally stable enzyme with a temperature optimum around 100°C and an apparent melting temperature of 105°C. The novel cellulase retains 64% of its activity after 4 hours of incubation at 95°C. The closest characterized homolog of AMOR_GH9A is Tf Cel9A, a processive endocellulase from the model thermophilic bacterium Thermobifida fusca (64.2% sequence identity). Direct comparison of AMOR_GH9A and Tf Cel9A revealed that AMOR_GH9A possesses higher activity on soluble and amorphous substrates (phosphoric acid swollen cellulose, konjac glucomannan) and has an ability to hydrolyse xylan that is lacking in Tf Cel9A.

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Cellulases from Thermophiles Found by Metagenomics

Cited by 50 publications

References 177 publications

Metagenomic applications in exploration and development of novel enzymes from nature: a review

Metagenomic applications in exploration and development of novel enzymes from nature: a review

Microbial diversity analysis and screening for novel xylanase enzymes from the sediment of the Lobios Hot Spring in Spain

Identification and characterization of a hyperthermophilic GH9 cellulase from the Arctic Mid-Ocean Ridge vent field

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