Endoglucanases are increasingly applied in agricultural and industrial applications as a key biocatalyst for cellulose biodegradation. However, the low performance in extreme conditions seriously challenges the enzyme’s commercial utilization. To obtain endoglucanases with substantially improved activity and thermostability, structure-based rational design was carried out based on the Chaetomium thermophilum β-1,4-endoglucanase CTendo45. In this study, five mutant enzymes were constructed by substitution of conserved and noncatalytic residues using site-directed mutagenesis. Mutants were constitutively expressed in Pichia pastoris, purified, and ultimately tested for enzymatic characteristics. Two single mutants, Y30F and Y173F, increased the enzyme’s specific activity 1.35- and 1.87-fold using carboxymethylcellulose sodium (CMC-Na) as a substrate, respectively. Furthermore, CTendo45 and mutants exhibited higher activity towards β-D-glucan than that of CMC-Na, and activities of Y173F and Y30F were also increased obviously against β-D-glucan. In addition, Y173F significantly improved the enzyme’s heat resistance at 80 °C and 90 °C. More interestingly, the double mutant Y30F/Y173F obtained considerably higher stability at elevated temperatures but failed to inherit the increased catalytic efficiency of its single mutant counterparts. This work gives an initial insight into the biological function of conserved and noncatalytic residues of thermostable endoglucanases and proposes a feasible path for the improvement of enzyme redesign proposals.
Information on seed germination and emergence ecology of Aegilops tauschii is scant, despite it being a widespread invasive weed in China. We conducted this study to determine the effects of various factors on seed germination and seedling emergence in three A. tauschii populations. Seeds germinated across a wide range of temperatures (5–35°C), with germination of over 90% at 15–20°C. Germination was completely inhibited when dry seeds were exposed to a temperature of 160°C for 5 min; a similar response was observed for pre‐soaked seeds at 100°C. Light was neither required for nor inhibited germination. Germination was not significantly affected by pH. Aegilops tauschii was relatively tolerant to low osmotic potential and high salt stress: over 80% of seeds germinated at −0.3 MPa, and all three populations germinated in the presence of 400 mM salt (NaCl) although salt tolerance varied among the populations. Seeds buried at depths of 1–3 cm emerged well, but emergence was completely inhibited at depths greater than 8 cm. The addition of maize straw caused a linear reduction in seedling emergence, although the rate of reduction varied among the populations. The results of this study have contributed to understanding the requirements of A. tauschii germination and emergence and optimising an integrated management system for this weed in Huang–Huai–Hai Plain of China. In addition, our study provides data for development of models to predict the geographical distribution of this weed.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.
Copyright © 2025 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.