Improvement of Trichoderma reesei xylanase II thermal stability by serine to threonine surface mutations

Ayadi, Dorra; Sayari, Aida Hmida; Hlima, Hajer Ben; Mabrouk, Sameh; Mezghani, Monia; Béjar, Samír

doi:10.1016/j.ijbiomac.2014.08.014

Cited by 32 publications

(10 citation statements)

References 50 publications

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“…The previous research found that the mutation from Ser to Arg and Asn to Ser were the most common mutations. So far as the consequences of these mutations, GH11 xylanases enhanced their catalytic activity and thermostability (Satyanarayana, 2013; Ayadi et al, 2015). Apart from that, the mutations (T11Y, N12H, N13D, F15Y, Y16F, S62T, S138T, S144C, N198D, A217V, and L49P) were discovered to be involved in conferring thermostability and catalytic activity on xylanases, including SoxB, SlxB, AnxA, AnxB, and BaxA (Zhang et al, 2010; Wang et al, 2012, 2015; Xu et al, 2016).…”

Section: Resultsmentioning

confidence: 99%

Laboratory Evolution of GH11 Endoxylanase Through DNA Shuffling: Effects of Distal Residue Substitution on Catalytic Activity and Active Site Architecture

Liu

Rehman

et al. 2019

Front. Bioeng. Biotechnol.

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Endoxylanase with high specific activity, thermostability, and broad pH adaptability is in huge demand. The mutant library of GH11 endoxylanase was constructed via DNA shuffling by using the catalytic domain of Bacillus amyloliquefaciens xylanase A (BaxA) and Thermomonospora fusca TF xylanase A (TfxA) as parents. A total of 2,250 colonies were collected and 756 of them were sequenced. Three novel mutants (DS153: N29S, DS241: S31R and DS428: I51V) were identified and characterized in detail. For these mutants, three residues of BaxA were substituted by the corresponding one of TfxA_CD. The specific activity of DS153, DS241, and DS428 in the optimal condition was 4.54, 4.35, and 3.9 times compared with the recombinant BaxA (reBaxA), respectively. The optimum temperature of the three mutants was 50°C. The optimum pH for DS153, DS241, and DS428 was 6.0, 7.0, and 6.0, respectively. The catalytic efficiency of DS153, DS241, and DS428 enhanced as well, while their sensitivity to recombinant rice xylanase inhibitor (RIXI) was lower than that of reBaxA. Three mutants have identical hydrolytic function as reBaxA, which released xylobiose–xylopentaose from oat spelt, birchwood, and beechwood xylan. Furthermore, molecular dynamics simulations were performed on BaxA and three mutants to explore the precise impact of gain-of-function on xylanase activity. The tertiary structure of BaxA was not altered under the substitution of distal residues (N29S, S31R, and I51V); it induced slightly changes in active site architecture. The distal impact rescued the BaxA from native conformation (“closed state”) through weakening interactions between “gate” residues (R112, N35 in DS241 and DS428; W9, P116 in DS153) and active site residues (E78, E172, Y69, and Y80), favoring conformations with an “open state” and providing improved activity. The current findings would provide a better and more in-depth understanding of how distal single residue substitution improved the catalytic activity of xylanase at the atomic level.

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Section: Resultsmentioning

confidence: 99%

Laboratory Evolution of GH11 Endoxylanase Through DNA Shuffling: Effects of Distal Residue Substitution on Catalytic Activity and Active Site Architecture

Liu

Rehman

et al. 2019

Front. Bioeng. Biotechnol.

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“…Similarly, four cumulative mutations of serine/threonine with arginine residues have significantly enhanced the thermostability of metagenomic xylanase at 90 • C . The introduction of at least five arginine residues in endoxylanase of Trichoderma reesei resulted in a shift in optimum temperature as well as pH (Turunen et al, 2002), while Ayadi et al (2015) obtained significant improvement in thermostability by replacing serine with threonine residues. Molecular modeling revealed that such mutations (S80T and S149T) assisted in hydrogen bonding and exhibited a packing effect (Ayadi et al, 2015).…”

Section: Molecular Attributes To Improve Extremophilic Endoxylanasesmentioning

confidence: 99%

Extremophilic Prokaryotic Endoxylanases: Diversity, Applicability, and Molecular Insights

Verma

2021

Front. Microbiol.

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Extremophilic endoxylanases grabbed attention in recent years due to their applicability under harsh conditions of several industrial processes. Thermophilic, alkaliphilic, and acidophilic endoxylanases found their employability in bio-bleaching of paper pulp, bioconversion of lignocellulosic biomass into xylooligosaccharides, bioethanol production, and improving the nutritious value of bread and other bakery products. Xylanases obtained from extremophilic bacteria and archaea are considered better than fungal sources for several reasons. For example, enzymatic activity under broad pH and temperature range, low molecular weight, cellulase-free activity, and longer stability under extreme conditions of prokaryotic derived xylanases make them a good choice. In addition, a short life span, easy cultivation/harvesting methods, higher yield, and rapid DNA manipulations of bacterial and archaeal cells further reduces the overall cost of the product. This review focuses on the diversity of prokaryotic endoxylanases, their characteristics, and their functional attributes. Besides, the molecular mechanisms of their extreme behavior have also been presented here.

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“…It utilizes the error-prone PCR or DNA shuffling technique in combination with the high-throughput screening method to continuously accumulate the dominant mutations with improved characteristics of the enzyme [5,11,12,13,14,15]. Rational design is conducted based on the understanding of the catalytic mechanism or the enzyme structure in which the stereo-structure can sometimes be predicted by protein homology modeling technique [16,17,18,19,20,21]. The key amino acids that may affect the enzyme properties can be chosen for site-directed mutagenesis, which includes the single site-directed mutation, multiple site-directed mutations, and saturation mutation.…”

Section: Introductionmentioning

confidence: 99%

Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes

et al. 2018

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The β-glycoside hydrolases (LXYL-P1−1 and LXYL-P1−2) from Lentinula edodes (strain M95.33) can specifically hydrolyze 7-β-xylosyl-10-deacetyltaxol (XDT) to form 10-deacetyltaxol for the semi-synthesis of Taxol. Our previous study showed that both the I368T mutation in LXYL-P1−1 and the T368E mutation in LXYL-P1−2 could increase the enzyme activity, which prompted us to investigate the effect of the I368E mutation on LXYL-P1−1 activity. In this study, the β-xylosidase and β-glucosidase activities of LXYL-P1−1I368E were 1.5 and 2.2 times higher than those of LXYL-P1−1. Most importantly, combination of I368E and V91S exerted the cumulative effects on the improvement of the enzyme activities and catalytic efficiency. The β-xylosidase and β-glucosidase activities of the double mutant LXYL-P1−1V91S/I368E were 3.2 and 1.7-fold higher than those of LXYL-P1−1I368E. Similarly, the catalytic efficiency of LXYL-P1−1V91S/I368E on 7-β-xylosyl-10-deacetyltaxol was 1.8-fold higher than that of LXYL-P1−1I368E due to the dramatic increase in the substrate affinity. Molecular docking results suggest that the V91S and I368E mutation might positively promote the interaction between enzyme and substrate through altering the loop conformation near XDT and increasing the hydrogen bonds among Ser91, Trp301, and XDT. This study lays the foundation for exploring the relationship between the structure and function of the β-glycoside hydrolases.

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Improvement of Trichoderma reesei xylanase II thermal stability by serine to threonine surface mutations

Cited by 32 publications

References 50 publications

Laboratory Evolution of GH11 Endoxylanase Through DNA Shuffling: Effects of Distal Residue Substitution on Catalytic Activity and Active Site Architecture

Laboratory Evolution of GH11 Endoxylanase Through DNA Shuffling: Effects of Distal Residue Substitution on Catalytic Activity and Active Site Architecture

Extremophilic Prokaryotic Endoxylanases: Diversity, Applicability, and Molecular Insights

Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes

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