Characterization of two thermophilic cellulases from Talaromyces leycettanus JCM12802 and their synergistic action on cellulose hydrolysis

Gu, Yuan; Zheng, Fu‐Chun; Wang, Yuan; Su, Xiaoyun; Bai, Yingguo; Yao, Bin; Huang, Huoqing; Luo, Huiying

doi:10.1371/journal.pone.0224803

Cited by 15 publications

(9 citation statements)

References 49 publications

(55 reference statements)

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“…The GH16 elongating β-transglycosylase from Paecilomyces thermophila (PtBgt16A), which also performed hydrolysis of mixed linkages, showed a pH optimum of 5.5 and a temperature optimum of 60 • C [18]. Additionally, a recent study showed the GH5A enzyme from Talaromyces leycettanus JCM12802 to have a high specific activity on a mixed-linkage substrate [19]. The enzyme showed a pH optimum of 3 and a temperature optimum of 75 • C. In contrast, Exg-D showed a broad pH optimum from 5.5 to 7 and a temperature optimum from 40 to 60 • C. The broad pH and temperature optima of Exg-D enzyme make it a potential candidate for the hydrolysis of licheninor β-glucan-containing biomass.…”

Section: Discussionmentioning

confidence: 99%

“…This distinction between GH16 enzymes and Exg-D emphasised the novelty of the mechanism employed by Exg-D during the hydrolysis of mixed-linkage oligosaccharides/substrates. In addition, Exg-D was also highly active on cello-oligosaccharides, and it employed the well-studied processive mechanism of exoglucanases [19].…”

Section: Discussionmentioning

confidence: 99%

“…Exoglucanases are known for hydrolysing their substrates in a processive manner, meaning that the enzyme moves along the backbone of cellulose or glucan, releasing cellobiose [9,18,19]. The processivity of the exoglucanases is linked to their tunnel-like-cleft active site, as demonstrated by Qin et al, [18].…”

Section: Discussionmentioning

confidence: 99%

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A Novel Dimeric Exoglucanase (GH5_38): Biochemical and Structural Characterisation towards its Application in Alkyl Cellobioside Synthesis

et al. 2020

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An exoglucanase (Exg-D) from the glycoside hydrolase family 5 subfamily 38 (GH5_38) was heterologously expressed and structurally and biochemically characterised at a molecular level for its application in alkyl glycoside synthesis. The purified Exg-D existed in both dimeric and monomeric forms in solution, which showed highest activity on mixed-linked β-glucan (88.0 and 86.7 U/mg protein, respectively) and lichenin (24.5 and 23.7 U/mg protein, respectively). They displayed a broad optimum pH range from 5.5 to 7 and a temperature optimum from 40 to 60 • C. Kinetic studies demonstrated that Exg-D had a higher affinity towards β-glucan, with a K m of 7.9 mg/mL and a k cat of 117.2 s −1 , compared to lichenin which had a K m of 21.5 mg/mL and a k cat of 70.0 s −1 . The circular dichroism profile of Exg-D showed that its secondary structure consisted of 11% α-helices, 36% β-strands and 53% coils. Exg-D performed transglycosylation using p-nitrophenyl cellobioside as a glycosyl donor and several primary alcohols as acceptors to produce methyl-, ethyl-and propyl-cellobiosides. These products were identified and quantified via thin-layer chromatography (TLC) and liquid chromatography-mass spectrometry (LC-MS). We concluded that Exg-D is a novel and promising oligomeric glycoside hydrolase for the one-step synthesis of alkyl glycosides with more than one monosaccharide unit.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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A Novel Dimeric Exoglucanase (GH5_38): Biochemical and Structural Characterisation towards its Application in Alkyl Cellobioside Synthesis

et al. 2020

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“…The recombinant enzyme was successfully expressed by P. pastoris, with a main product that shows a molecular size of about 50 kDa. The recombinant Plasmid Cel6A selected was linearized with the enzyme BamHIO recombinant plasmid, pLIPI-TrCel6A, carrier of the T. r e e s e i Cel6A gene (TrCel6A) integrated into the genome of P. pastoris SMD1168H [50].…”

Section: Recombinant Fungal Cellulases Produced By Different Expression Systemsmentioning

confidence: 99%

Recombinant Fungal Cellulases for the Saccharification of Sugarcane Bagasse

Benevides¹,

Assis²,

Vitor³

et al. 2022

Biodegradation Technology of Organic and Inorganic Pollutants

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Cellulases are important enzymes in cellulose degradation that occurs in nature, this degradation involves a system of extracellular multienzymes and have wide application. The construction of a high-quality system for the production of these enzymes is important for its application in the process of saccharification of biomass involved in the biofuel production process. Several species of fungi are capable of synthesizing and secreting high amounts of cellulase, most studies with fungal species use linearized plasmid, since these are encompassed to chromosomal DNA, improving its stability and expression efficiency. Advances in the production of recombinant enzymes focus on the search for industrially viable microorganisms capable of producing enzymes under various conditions, expressing them in a highly efficient manner, aiming at the synthesis of several copies of genes and a strong promoter. To resay these restrictions, molecular biology combined with recombinant DNA technology is a viable tool in enzymatic production. In subsequent topics, the production of endoglucanases, exoglucanases and β-glucosidase of fungi cloned in Escherichia coli, Pichia pastoris and other different expression systems will be addressed.

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“…Talaromyces leycettanus JCM12802 is a typical thermophilic fungus with an optimum temperature for growth at 40 ℃, producing a wide range of thermostable glycoside hydrolases, such as cellulase, β-glucosidase, and mannanase [ 23 – 25 ]. In this study, a novel thermostable glucoamylase Tl Ga15B from T. leycettanus JCM12802 was expressed and characterized.…”

Section: Introductionmentioning

confidence: 99%

Improvement of thermostability and catalytic efficiency of glucoamylase from Talaromyces leycettanus JCM12802 via site-directed mutagenesis to enhance industrial saccharification applications

Tong

Zheng

Wang

et al. 2021

Biotechnol Biofuels

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Background Glucoamylase is an important industrial enzyme in the saccharification of starch into glucose. However, its poor thermostability and low catalytic efficiency limit its industrial saccharification applications. Therefore, improving these properties of glucoamylase is of great significance for saccharification in the starch industry. Results In this study, a novel glucoamylase-encoding gene TlGa15B from the thermophilic fungus Talaromyces leycettanus JCM12802 was cloned and expressed in Pichia pastoris. The optimal temperature and pH of recombinant TlGa15B were 65 ℃ and 4.5, respectively. TlGa15B exhibited excellent thermostability at 60 ℃. To further improve thermostability without losing catalytic efficiency, TlGa15B-GA1 and TlGa15B-GA2 were designed by introducing disulfide bonds and optimizing residual charge–charge interactions in a region distant from the catalytic center. Compared with TlGa15B, mutants showed improved optimal temperature, melting temperature, specific activity, and catalytic efficiency. The mechanism underlying these improvements was elucidated through molecular dynamics simulation and dynamics cross-correlation matrices analysis. Besides, the performance of TlGa15B-GA2 was the same as that of the commercial glucoamylase during saccharification. Conclusions We provide an effective strategy to simultaneously improve both thermostability and catalytic efficiency of glucoamylase. The excellent thermostability and high catalytic efficiency of TlGa15B-GA2 make it a good candidate for industrial saccharification applications.

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Characterization of two thermophilic cellulases from Talaromyces leycettanus JCM12802 and their synergistic action on cellulose hydrolysis

Cited by 15 publications

References 49 publications

A Novel Dimeric Exoglucanase (GH5_38): Biochemical and Structural Characterisation towards its Application in Alkyl Cellobioside Synthesis

A Novel Dimeric Exoglucanase (GH5_38): Biochemical and Structural Characterisation towards its Application in Alkyl Cellobioside Synthesis

Recombinant Fungal Cellulases for the Saccharification of Sugarcane Bagasse

Improvement of thermostability and catalytic efficiency of glucoamylase from Talaromyces leycettanus JCM12802 via site-directed mutagenesis to enhance industrial saccharification applications

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