Screening, cloning, enzymatic properties of a novel thermostable cellulase enzyme, and its potential application on water hyacinth utilization

Abstract

Cellulose is the cheapest, natural, renewable organic substance that is used as a carbon source in various elds. Water hyacinth, an aquatic plant rich in cellulose, is often used as a raw material in fuel production. However, natural cellulase can be hardly used in industrial production on account of its low thermal stability and activity. In this study, a metagenomic library was constructed. Then a new cellulase gene, cel1029, was screened by Congo red staining and expressed in the prokaryotic system. Enzymatic properties of Cel1029 were explored, including optimum temperature and pH, thermal and pH stability, and tolerance against organic solvents, metal ions and salt solutions. Finally, its ability of degrading water hyacinth was identi ed and evaluated. Cel1029 displayed high homology with endoglucanase in the glycoside hydrolase family 5 (GH5) and had high stability across a broad temperature range. More than 86% of its enzymatic activities were retained between 4 and 60°C after 24 h of incubation. Singlefactor analysis and orthogonal design were further conducted to determine the optimal conditions for the highest reducing sugar yield of water hyacinth. Interestingly, Cel1029 e ciently transformed water hyacinth with a reducing sugar yield of 430.39 mg/g in 22 h. These ndings may open the door for signi cant industrial applications of a novel GH5 cellulase (NCBI Reference Sequence: MK051001, Cel1029) and help identify more e cient methods to degrade cellulose-rich plants.

Effects of novel cellulase (Cel 906) and probiotic yeast fermentation on antioxidant and anti-inflammatory activities of vine tea (Ampelopsis grossedentata)

“…Cellulase can hydrolyze the walls of plant cells, increase the extraction rate of effective substances in plant cells and improve the utilization rate ( Rostami, and Gharibzahedi, 2017 ; Zhao et al, 2021 ). As a result, more effective substances enter the fermentation system and can convert macromolecular substances into small-molecular substances that can be easily absorbed by the skin ( Sun et al, 2020 ).…”

Expression, characterization, and immobilization of a novel SGNH esterase Est882 and its potential for pyrethroid degradation

“…A new esterase gene was isolated from the soil metagenomic library in Junggar Basin, Xinjiang ( Zhao et al, 2021 ). The esterase gene is in full length of 882 bp and encodes a protein of 294 amino acids (named Est882).…”

“…The sample was taken from 5 to 10 cm under the surface, sealed in a sterile bag, and preserved at −20°C until DNA extraction. Total genomic DNA from the soil sample was extracted according to a reported method ( Zhao et al, 2021 ). In the metagenomic library of soil from Junggar Basin, the gene that express the protein with esterase activity was successfully extracted using a genomic DNA extraction kit (Omega, USA).…”

Pretreatment and composting technology of agricultural organic waste for sustainable agricultural development

“…The components in lignocellulose form a lignocellulosic matrix through covalent or non-covalent bonds, which gives it a strong natural resistance to hydrolysis by hydrolytic enzymes [ 43 ]. This disobedience needs to be overcome by some pretreatment to promote the hydrolysis of lignocellulose [ 44 ]. The primary purpose of pretreatment is to promote lignin hydrolysis, disrupt cellulose crystal structure, increase its surface area, and improve hydrolysis efficiency [ 45 ].…”