Cloning, expression and characterization of a cold-adapted endo-1, 4-β-glucanase from Citrobacter farmeri A1, a symbiotic bacterium of Reticulitermes labralis

Bai, Xi; Yuan, Xianjun; Wen, Aiyou; Li, Junfeng; Bai, Yunfeng; Shao, Tao

doi:10.7717/peerj.2679

Cited by 13 publications

(10 citation statements)

References 24 publications

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“…The optimum temperature of EG-PY2 was lower than that of CelX isolated from a deep sea-bacterium Pseudoalteromonas sp. and of EglC isolated from a symbiotic bacterium Citrobacter farmeri (Zeng et al 2006;Bai et al 2016). The cold activity pattern of EG-PY2 did not change markedly between 20 and 50 o C, which is similar to those reported for reasons of the endoglucanases Cel9P, EG5C and PgluE8 from Paenibacillus sp.…”

Section: Properties Of Eg-py2supporting

confidence: 70%

A cold-active acidophilic endoglucanase of Paenibacillus sp. Y2 isolated from soil in an alpine region

Lee

Seo

et al. 2017

JABC

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A cellulolytic strain Y2 was isolated from soil obtained in the Canadian Alpine region. The isolate was identified as Paenibacillus sp. Y2 by 16S rRNA sequencing. When grown in LB medium supplemented with carboxymethyl-cellulose (CMC), CMCase production increased to 122.0% of that observed in LB without CMC. Culture supernatant was concentrated by ultrafiltration and 80% ammonium sulfate precipitates were separated by HiTrap Q and CHT-II chromatography. The purified enzyme (EG-PY2) showed a homogeneous single band and the molecular mass was estimated to be 38 kDa by SDS-PAGE. Optimum pH and temperature of the enzyme were 4.5 and 30 o C, respectively. The half-life of enzyme activity at 50 was 140.7 min, but the enzyme was drastically inactivated within 5 min at 55 , and EDTA. The enzyme was activated to 211.5% in the presence of 0.5 M of NaCl and greatly tolerant to 3.15 M of NaCl. The enzyme showed 2.98 times higher β-glucanase activity than CMCase activity. Based on these results, it can be concluded that EG-PY2 is an acidophilic, cold-active, and halotolerant endoglucanase. The authors suggest it is considered to be useful for various industrial applications, such as, fruit juice clarification, acidic deinking processes, high-salt food processing, textile and pulp industries, and for biofuel production from seaweeds.

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Section: Properties Of Eg-py2supporting

confidence: 70%

A cold-active acidophilic endoglucanase of Paenibacillus sp. Y2 isolated from soil in an alpine region

Lee

Seo

et al. 2017

JABC

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“…Further, we conducted the SDS-PAGE analysis under denaturing conditions, and the molecular weight of the purified recombinant BpEG was estimated to be 60 kDa ( Figure 3A). The molecular weights of the endo-1,4-β-glucanases from other species, namely Rhizopus oryzae (Murashima et al, 2002), Citrobacter farmeri A1 (Bai et al, 2016), and Ganoderma lucidum (Liu et al, 2016), were previously reported.…”

Section: Heterologous Expression Of Recombinant Bpegmentioning

confidence: 74%

“…The sequencing analysis of the above-obtained recombinant BpEG fragments using an automatic protein sequencer showed us the conserved domains AIRVYLWAGM and VPGLGVTLLP. These conservative fragments exhibited high similarity with the endo-1,4-β-glucanases sequences of E. coli (Park and Yun, 1999) and Citrobacter farmeri A1 (Bai et al, 2016), which belong to GH 8.…”

Section: Western Blot Analysis and Identification Of The Partial Peptmentioning

confidence: 92%

“…The optimum pH of the recombinant BpEG protein showed comparable results with the previously reported endo-β-1,4 glucanases from Rhizopus oryzae (Murashima et al, 2002), Eisenia foetida (Ueda et al, 2014), and G. lucidum (Liu et al, 2016), respectively. However, another study reported that endo-1,4-β-glucanases isolated from the Citrobacter farmeri A1 showed an optimum pH of 3.5-7.5 (Bai et al, 2016). The activity of the recombinant BpEG was also determined at various temperatures, ranging from 10-60 • C at an optimum pH of 6.0.…”

Section: Properties Of Recombinant Bpegmentioning

confidence: 99%

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A Novel Cold-Adaptive Endo-1,4-β-Glucanase From Burkholderia pyrrocinia JK-SH007: Gene Expression and Characterization of the Enzyme and Mode of Action

Chen

Kameshwar

et al. 2020

Front. Microbiol.

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The efficient industrial conversion of plant-derived cellulose to simple sugars and other value-added chemicals requires various highly stable and reactive enzymes. Industrial processes especially synchronous saccharification and fermentation (SSF)based production of cellulosic bio-ethanol require enzymes that are active at lower temperatures. In this study, we have identified, characterized, and expressed the cold-adaptive endo-1,4-β-glucanase (BpEG) isolated from the Burkholderia pyrrocinia JK-SH007. The analysis of the predicted amino acid sequence indicated that BpEG belongs to GH family 8. The BpEG without the signal peptide was cloned into the expression vector pET32a and significantly expressed in Escherichia coli BL21 (DE3) competent cells. The SDS-PAGE and Western blot analysis of BpEG revealed that the recombinant BpEG was approximately 60 kDa. Purified recombinant BpEG exhibited hydrolytic activity against carboxymethyl cellulose (CMC) and phosphoric acid swollen cellulose (PASC), but not crystalline cellulose and xylan substrates. High performance, anion exchange, chromatography-pulsed amperometric detector (HPAEC-PAD) analysis of the enzymatic products obtained from depolymerization of 1,4-β-linked biopolymers of different lengths revealed an interesting cutting mechanism employed by endoglucanases. The recombinant BpEG exhibited 6.0 of optimum pH and 35 • C of optimum temperature, when cultured with CMC substrate. The BpEG enzyme exhibited stable activity between pH 5.0 and 9.0 at 35 • C. Interestingly, BpEG retained about 42% of its enzymatic activity at 10 • C compared to its optimal temperature. This new cold-adaptive cellulase could potentially achieve synchronous saccharification and fermentation (SSF) making BpEG a promising candidate in the fields of biofuel, biorefining, food and pharmaceutical industries.

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“…Some of the cellulases previously reported and applied on the market were mesophilic and thermophilic enzymes [5][6][7][8][9][10][11][12], while a few cold-adapted cellulases have been studied, such as CaCel from Cryptopygus antarctica [13], CelG from Fibrobacter succinogenes S85 [14], CelM of Pseudomonas sp. MM15 [15], etc. Compared with medium-temperature cellulase, cold-active enzymes have a unique role in industrial production and food processing [16].…”

Section: Introductionmentioning

confidence: 99%

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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Cloning, expression and characterization of a cold-adapted endo-1, 4-β-glucanase from Citrobacter farmeri A1, a symbiotic bacterium of Reticulitermes labralis

Cited by 13 publications

References 24 publications

A cold-active acidophilic endoglucanase of Paenibacillus sp. Y2 isolated from soil in an alpine region

A cold-active acidophilic endoglucanase of Paenibacillus sp. Y2 isolated from soil in an alpine region

A Novel Cold-Adaptive Endo-1,4-β-Glucanase From Burkholderia pyrrocinia JK-SH007: Gene Expression and Characterization of the Enzyme and Mode of Action

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

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