A novel pH-stable, endoglucanase (JqCel5A) isolated from a salt-lake microorganism, Jonesia quinghaiensis

Lin, Ling; Liu, Xiaozhou; Zhou, Yating; Guan, Linyan; He, Jiajia; Huang, Weixiong

doi:10.1016/j.ejbt.2016.09.004

Cited by 16 publications

(16 citation statements)

References 32 publications

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“…Thus the endoglucanase CelRH5 is more active in slightly acidic conditions but is also quite active and stable under slightly alkaline conditions. While a number of other metagenome-derived endoglucanases from GH5 have also shown highest activity in pH range 4.5–7.0 (Feng et al, 2007; Alvarez et al, 2013; Martin et al, 2014; Lin et al, 2016), there are fewer examples of endoglucanases exhibiting activity over such a broad pH range (Lee et al, 2014; Garg et al, 2016; Lin et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a Novel Monospecific Endo-β-1,4-Glucanase Belonging to GH Family 5 From a Rhizosphere Metagenomic Library

et al. 2019

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Cellulases have a broad range of different industrial applications, ranging from food and beverages to pulp and paper and the biofuels area. Here a metagenomics based strategy was used to identify the cellulolytic enzyme CelRH5 from the rhizosphere. CelRH5 is a novel monospecific endo-β-1,4-glucanase belonging to the glycosyl hydrolase family 5 (GH5). Structural based modeling analysis indicated that CelRH5 is related to endo-β-1,4-glucanases derived from thermophilic microorganisms such as Thermotoga maritima , Fervidobacterium nodosum , and Ruminiclostridium thermocellum sharing 30-40% amino acid sequence identity. The molecular weight of the enzyme was determined as 40.5 kDa. Biochemical analyses revealed that the enzyme displayed good activity with soluble forms of cellulose as a substrate such as ostazin brilliant red hydroxyethyl cellulose (OBR-HEC), carboxymethylcellulose (CMC), hydroxyethyl cellulose (HEC), and insoluble azurine cross-linked hydroxyethylcellulose (AZCL-HEC). The enzyme shows highest enzymatic activity at pH 6.5 with high pH tolerance, remaining stable in the pH range 4.5–8.5. Highest activity was observed at 40°C, but CelRH5 is psychrotolerant being active and stable at temperatures below 30°C. The presence of the final products of cellulose hydrolysis (glucose and cellobiose) or metal ions such as Na + , K + , Li + , and Mg 2+ , as well as ethylenediaminetetraacetic acid (EDTA), urea, dithiothreitol (DTT), dimethyl sulfoxide (DMSO), 2-mercaptoethanol (2-ME) or glycerol, did not have a marked effect on CelRH5 activity. However, the enzyme is quite sensitive to the presence of 10 mM ions Zn 2+ , Ni 2+ , Co 2+ , Fe 3+ and reagents such as 1 M guanidine HCl, 0.1% sodium dodecyl sulfate (SDS) and 20% ethanol. Given that it is psychrotolerant and retains activity in the presence of final cellulose degradation products, metal ions and various reagents, which are common in many technological processes; CelRH5 may be potential suitability for a variety of different biotechnological applications.

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Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a Novel Monospecific Endo-β-1,4-Glucanase Belonging to GH Family 5 From a Rhizosphere Metagenomic Library

et al. 2019

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“…After that, the mixture was mixed thoroughly with 800 lL H 2 O and the absorbance of the mixture was measured at 540 nm. With glucose solution as the standard solution, an enzyme unit is defined as the amount of enzyme required to produce 1 lmol glucose per minute at 25 C [4].…”

Section: Characterization Of Enzyme Activitymentioning

confidence: 99%

“…Eight to nine ligand bonds involved in this mechanism can be linked to multiple secondary structures or protein domains to increase the stability of substrate binding to enzymes. It has also been reported that the mechanism underlying the reduction of the enzyme activity in the presence of Cu 2þ lies in its ability to bind the mercaptan groups of some amino acids or to interact with the carboxyl groups of some amino acids [4].…”

Section: Effect Of Temperature Ph Metal Ions and Some Reagents On Tmentioning

confidence: 99%

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Enzymatic identification and functional sites study of a novel cold-active cellulase (MkCel5) from Microbacterium kitamiensea

Wang

Zhou

et al. 2019

Biotechnology & Biotechnological Equipment

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A strain of Microbacterium kitamiense capable of decomposing cellulose was obtained from the soil of Shigatse region in Tibet. Low temperature and cellulase gene MkCel5 was cloned and sequenced. The size of the open-reading frame was 1449 bp; it was predicted to encode a polypeptide with 482 residues with a molecular weight of 51.38 kDa. Sequence analysis indicated that it belongs to the glycoside hydrolase family 5 (GH5), which has high homology with reported endoglucanases. The predicted protein structure had a typical carbohydrate-binding domain and a catalytic domain of cellulase. Site mutagenesis experiments showed that the two glutamates at positions 312 and 404 were crucial in the catalytic process. MkCel5 was identified as a cold-adaptive and neutral enzyme with a maximum activity at 25 C and pH 7.0. These properties suggested that MkCel5 has potential for application in the cellulose processing industry requiring low temperature.

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“…However, if, in the future, ethanol from lignocellulosic biomass through the enzymatic route becomes a major transportation fuel, these enzymes may occupy the largest volume of industrial enzymes (SINGHANIA et al, 2013). In particular, endoglucanases have a great potential application in textile and detergent industries, paper recycling, juice extraction, animal feed additives and renewable energy (LIN et al, 2016). The main disadvantage is that cellulases have high production costs.…”

Section: Introductionmentioning

confidence: 99%

Endoglucanase production by endophytic fungi isolated from Vitis labrusca L. with peanut hull and sawdust as substrates

Felber¹,

Specian²,

Orlandelli³

et al. 2019

Biosci. J.

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Endoglucanases are enzymes widely employed in different industrial fields, albeit with high production costs. Studies on new microbial sources and low-cost substrates are highly relevant, including those on agro-industrial. Current analysis evaluates peanut hull (PH) and sawdust (SD) as substrates for submerged cultures of 14 endophytic fungi isolated from grapevine (Vitis labrusca L.) cultivars Bordô and Concord. Endophytes were grown on a carboxymethylcellulose (CMC) medium and the cup plate assay showed that eight strains (belonging to genera Cochliobolus, Diaporthe, Fusarium and Phoma) had positive results: enzymatic halos ranged from 10.8±0.02 to 15.5±0.07 mm in diameter. Diaporthe sp. strains (GenBank accession codes KM362392, KM362368 and KM362378) and Fusarium culmorum KM362384 were highlighted as the most promising sources. Further, PH and SD as substrates for the fermentation of these fungi were evaluated by the cup plate assay and endoglucanase activity assay. Highest halo diameters were obtained for Diaporthe sp. KM362392: 16.1±0.01 mm (CMC), 14.5±0.01 mm (PH) and 14.7±0.03 mm (SD). The fungus also presented the highest levels of endoglucanase activity: analysis of variance revealed that CMC (3.52±0.98 µmol/min), PH (2.93±0.23 µmol/min) and SD (3.26±0.38 µmol/min) were similarly efficient as substrates. Results deepen knowledge on V. labrusca endophytes that may be endoglucanase sources, even though further optimizations in submerged cultures with PH and SD should be undertaken to increase the enzymatic production from these wastes.

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A novel pH-stable, endoglucanase (JqCel5A) isolated from a salt-lake microorganism, Jonesia quinghaiensis

Cited by 16 publications

References 32 publications

Biochemical Characterization of a Novel Monospecific Endo-β-1,4-Glucanase Belonging to GH Family 5 From a Rhizosphere Metagenomic Library

Biochemical Characterization of a Novel Monospecific Endo-β-1,4-Glucanase Belonging to GH Family 5 From a Rhizosphere Metagenomic Library

Enzymatic identification and functional sites study of a novel cold-active cellulase (MkCel5) from Microbacterium kitamiensea

Endoglucanase production by endophytic fungi isolated from Vitis labrusca L. with peanut hull and sawdust as substrates

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