Gene cloning, expression, immobilization and characterization of endo-xylanase from Geobacillus sp. TF16 and investigation of its industrial applications

Cakmak, Ummuhan; Ertunga, Nagihan Saglam

doi:10.1016/j.molcatb.2017.01.016

Cited by 23 publications

(15 citation statements)

References 46 publications

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“…XynDZ5 exhibits a preference toward oatspelt xylan compared to different xylans, with K M and k cat values equal to 25 g/L and 36 s −1 , respectively. Although the K M values for birchwood and beechwood xylans were relatively higher they remained within the range of 25 to 40 g/L, magnitude of value that is usually attributed to fungal xylanases (Liao et al, 2014), but are significantly higher than the K M values reported for thermophilic xylanases of bacterial origin (Cakmak and Ertunga, 2016).…”

Section: -80mentioning

confidence: 68%

XynDZ5: A New Thermostable GH10 Xylanase

et al. 2020

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Xylanolytic enzymes have a broad range of applications in industrial biotechnology as biocatalytic components of various processes and products, such as food additives, bakery products, coffee extraction, agricultural silage and functional foods. An increasing market demand has driven the growing interest for the discovery of xylanases with specific industrially relevant characteristics, such as stability at elevated temperatures and in the presence of other denaturing factors, which will facilitate their incorporation into industrial processes. In this work, we report the discovery and biochemical characterization of a new thermostable GH10 xylanase, termed XynDZ5, exhibiting only 26% amino acid sequence identity to the closest characterized xylanolytic enzyme. This new enzyme was discovered in an Icelandic hot spring enrichment culture of a Thermoanaerobacterium species using a recently developed bioinformatic analysis platform. XynDZ5 was produced recombinantly in Escherichia coli, purified and characterized biochemically. This analysis revealed that it acts as an endo-1,4β-xylanase that performs optimally at 65-75 • C and pH 7.5. The enzyme is capable of retaining high levels of catalytic efficiency after several hours of incubation at high temperatures, as well as in the presence of significant concentrations of a range of metal ions and denaturing agents. Interestingly, the XynDZ5 biochemical profile was found to be atypical, as it also exhibits significant exo-activity. Computational modeling of its three-dimensional structure predicted a (β/α) 8 TIM barrel fold, which is very frequently encountered among family GH10 enzymes. This modeled structure has provided clues about structural features that may explain aspects of its catalytic performance. Our results suggest that XynDZ5 represents a promising new candidate biocatalyst appropriate for several high-temperature biotechnological applications in the pulp, paper, baking, animal-feed and biofuel industries.

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Section: -80mentioning

confidence: 68%

XynDZ5: A New Thermostable GH10 Xylanase

et al. 2020

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“…TF16 displayed an increase in optimum temperature from 55 to 65°C and shift in the pH value from 6.0 to 8.5 as compared to the free enzyme. The enzyme was found to be effective in release of the reducing sugars from juice and poultry feed and oven spring in bakery [99]. The β-xylosidase of the extremely thermophilic G. thermodenitrificans is optimally active at 60°C and pH 7.0 and it catalyzes transxylosylation reactions in the presence of alcohols as acceptors.…”

Section: Production and Applications Of Thermostable Enzymes Of Geobamentioning

confidence: 99%

GeobacillusBacteria: Potential Commercial Applications in Industry, Bioremediation, and Bioenergy Production

Новик¹,

Savich²,

Meerovskaya³

2019

Growing and Handling of Bacterial Cultures

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The genus Geobacillus is represented by obligately thermophilic bacteria able to grow in the temperature range of 35-75°C. They are modest bacteria isolated from various sources on routine media such as nutrient agar. Originally classified as representatives of Bacillus, the species of Geobacillus were established in 2001 as a new genus. However, sequence similarity between all species indicates that at least some species need to be reclassified at the genus level. In addition to 16S rRNA, housekeeping genes, 16S-23S rRNA gene internal transcribed spacer, and repetitive sequences can be used in classification and identification of thermophilic bacteria. The ability to survive and grow at high temperatures as well as utilization and synthesis of a wide range of compounds makes these bacteria and their products attractive for use in various spheres: food, paper, biotechnology industries, medicine, bioremediation, etc. A broad spectrum of applications arouses increased interest in the study of physiological and biochemical characteristics and triggers emergence of new usage areas for Geobacillus, such as bioenergy. The growing demand for energy leads to the development of alternative technologic options. Geobacillus species demonstrated the ability to generate or enhance productivity of important sources of bioenergy such as ethanol, isobutanol, 2,3-butanediol, biodiesel, and biogas.

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“…12 All experiments were performed in triplicate and the results presented are the mean of three values. The MINITAB version 16 statistical software was used for all analysis 49 and the standard deviation was within ± 5%.…”

Section: Effect Of Some Metal Ions and Organic Solventsmentioning

confidence: 99%

Site-directed mutagenesis and characterization of recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4

Tuncay¹,

Çolak²,

Akatin³

et al. 2019

Rev.Roum.Chim.

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Many toxic insecticides used worldwide are organophosphates (OPs) derivatives. Phosphotriesterase (PTE) has been rewarding to protect against OP poisoning in vivo or in vitro, associated with advanced catalytic efficiency and stereoselectivity toward the hydrolysis of OPs. Phosphotriesterase homology protein (PHP) exhibits high sequence identity and similarity to PTE. In this study, site-directed mutagenesis on recombinant Geobacillus caldoxylosilyticus TK4 PHP (TK4PHP) was performed for improving the existing esterase activity, even gaining a new PTE activity. After eliminating the deficiencies in the recombinant TK4PHP gene, mutant proteins were purified and characterized biochemically. Considering all the data obtained, it was determined that the major sequence differences between PTE and TK4PHP were removed by three site-directed mutations. However the mutant TK4PHPs did not have PTE activity, it was informed that mutant esterases were more resistance to some metal ions and organic solvents and more thermal stable when it was compared with recombinant type.

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Gene cloning, expression, immobilization and characterization of endo-xylanase from Geobacillus sp. TF16 and investigation of its industrial applications

Cited by 23 publications

References 46 publications

XynDZ5: A New Thermostable GH10 Xylanase

XynDZ5: A New Thermostable GH10 Xylanase

GeobacillusBacteria: Potential Commercial Applications in Industry, Bioremediation, and Bioenergy Production

Site-directed mutagenesis and characterization of recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4

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