Characterization of a GH family 8 β-1,3-1,4-glucanase with distinctive broad substrate specificity from Paenibacillus sp. X4

Na, Han Beur; Jung, Won Kyeong; Jeong, Younhee; Kim, Hee Jung; Kim, Sung Kyum; Kim, Jungho; Yun, Han Dae; Lee, Jung-Kul; Kim, Hoon

doi:10.1007/s10529-014-1724-x

Cited by 23 publications

(15 citation statements)

References 29 publications

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“…These Burkholderia 1,4-β-glucanases all belonging to the glycoside hydrolase (GH) family 8 suggest that BpEG might also belong to the GH 8 family. Six strictly conserved residues of GH8 Glu 83, Asp144, Pro209, Trp230, Glu271, and Arg278 (Figure 1) were found in the catalytic module of BpEG according to the multiple sequence alignment (Figure 2) as described by Adachi et al (2004) and Na et al (2015). The amino acids sequencing analysis of BpEG using the ClustalW program revealed the conservative motif ASDADLWIAYALVEAGRLW (Figure 2).…”

Section: Cloning and Sequence Analysis Of Bpegmentioning

confidence: 99%

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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Section: Cloning and Sequence Analysis Of Bpegmentioning

confidence: 99%

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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“…Following that, PgluE8 showed 14.89, 3.68 and 5–25 % amino acid sequence identity with GH8 β-1,3-1,4-glucanase from Paenibacillus sp. X4 (Na et al 2015 ), GH16 β-1,3-1,4-glucanase from Paenibacillus sp. S09 (Cheng et al 2014 ), and with other endoglucanase, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Cellulose is considered to be the most abundant renewable resource (Xiang et al 2014 ). β-Glucans, which are homopolymers of d -glucose linked by β-glycosidic bonds, commonly exist as cellulose in plants, the bran of cereal grains, and the cell walls of yeast, fungi, and bacteria (Na et al 2015 ). Cellulose is solubilized by cellulases, which can be classified into the following three categories per catalytic mechanism: endoglucanase (E.C.3.2.1.4), exoglucanase (E.C.3.2.1.91) and β-glucosidase (E.C.3.2.1.21) (Javed et al 2009 ).…”

Section: Introductionmentioning

confidence: 99%

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NaCl-, protease-tolerant and cold-active endoglucanase from Paenibacillus sp. YD236 isolated from the feces of Bos frontalis

et al. 2016

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Bos frontalis, which consumes bamboo and weeds, may have evolved unique gastrointestinal microorganisms that digest cellulase. A Paenibacillus sp. YD236 strain was isolated from B. frontalis feces, from which a GH8 endoglucanase gene, pglue8 (1107 bp, 54.5 % GC content), encoding a 368-residue polypeptide (PgluE8, 40.4 kDa) was cloned. PgluE8 efficiently hydrolyzed barley-β-d-glucan followed by CMC-Na, soluble starch, laminarin, and glucan from black yeast optimally at pH 5.5 and 50 °C, and retained 78.6, 41.6, and 34.5 % maximum activity when assayed at 20, 10, and 0 °C, respectively. Enzyme activity remained above 176.6 % after treatment with 10.0 mM β-mercaptoethanol, and was 83.0, 78, and 56 % after pre-incubation in 30 % (w/v) NaCl, 16.67 mg/mL trypsin, and 160.0 mg/mL protease K, respectively. Cys23 and Cys364 residues were critical for PgluE8 activity. pglue8, identified from B. frontalis feces for the first time in this study, is a potential alternative for applications including food processing, washing, and animal feed preparation.

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“…The soil was obtained near Takakkaw Falls in the Canadian Alpine region (Na et al 2015). After serial dilutions of soil, supernatants were spread on Luria-Bertani (LB) agar plates.…”

Section: Isolation Of Cellulolytic Bacteria From Soilmentioning

confidence: 99%

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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Characterization of a GH family 8 β-1,3-1,4-glucanase with distinctive broad substrate specificity from Paenibacillus sp. X4

Cited by 23 publications

References 29 publications

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

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

NaCl-, protease-tolerant and cold-active endoglucanase from Paenibacillus sp. YD236 isolated from the feces of Bos frontalis

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

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