Cellulase Assays

Zhang, Y. H. Percival; Hong, Jiong; Ye, Xinhao

doi:10.1007/978-1-60761-214-8_14

Cited by 166 publications

(107 citation statements)

References 20 publications

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“…FT-IR analysis was conducted using a Perkin-Elmer FTIR spectrum-400 spectrometer (Perkin-Elmer Inc., Wellesley, MA), and the spectra were obtained in the range of 500 to 4000 cm -1 . The endoglucanase assay was performed with slight modification of the method reported by Zhang et al (2009). The reaction mixture contained 200 µL of the crude enzyme preparation with 200 µL of 2% CMC (medium viscosity) in 0.05 M phosphate buffer (pH 7.0).…”

Section: Methodsmentioning

confidence: 99%

Effects of Pretreatment of Single and Mixed Lignocellulosic Substrates on Production of Endoglucanase by Bacillus aerius S5.2

Oke¹,

Ishola²,

Taherzadeh³

et al. 2016

BioResources

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A mixed substrate (MS) comprising oil palm empty fruit bunch (EFB), oil palm frond (OPF), and rice husk (RH) was evaluated for endoglucanase production by Bacillus aerius S5.2. Effects of sulphuric acid, sodium hydroxide, N-methylmorpholine-N-oxide (NMMO), and hydrothermal pretreatments on endoglucanase production were investigated. Endoglucanase production by B. aerius on the untreated (0.677 U/mL) and pretreated MS (0.305 -0.630 U/mL) was generally similar, except that the acid (0.305 U/mL) and hydrothermal (0.549 U/mL) pretreatments that were more severe consequently produced significantly lower titres. Alkali pretreatment supported the highest enzyme production (0.630 U/mL) among all pretreatments that were studied. When endoglucanase production on the alkali-pretreated MS and single substrates (SS) was compared, alkali-pretreated EFB produced a titre (0.655 U/mL) similar to the MS, and this was significantly higher than titres recorded on OPF (0.504 U/mL) and RH (0.525 U/mL). Lower enzyme production was found to be consistent with higher pretreatment severity and greater removal of amorphous regions in all the pretreatments. Furthermore, combining the SS showed no adverse effects on endoglucanase production.

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Section: Methodsmentioning

confidence: 99%

Effects of Pretreatment of Single and Mixed Lignocellulosic Substrates on Production of Endoglucanase by Bacillus aerius S5.2

Oke¹,

Ishola²,

Taherzadeh³

et al. 2016

BioResources

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“…A wide range of wavelengths has been used to determine the reducing sugars using the broadly defined DNS method (Stellmach, 1992;Zhang et al, 2009). Different measurement wavelengths resulted in different OD values, and the OD value was used to evaluate the cellulase activity, thus, as OD values shift, so will cellulase activity results.…”

Section: Optimization Of Measuring the Wavelength For Cellulase Activitymentioning

confidence: 99%

Characteristics of cellulase in cellulose-degrading bacterium strain Clostridium straminisolvens (CSK1)

Wang¹,

Hua²,

Wang³

et al. 2017

Afr. J. Microbiol. Res.

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Microbial degradation of biomass was considered as a clean and highly efficient approach to produce bioenergy, as it could mitigate the urgent demand for limited petroleum and natural gas. A thermophilic microbial consortium (MC1) was proved a high efficiency cellulose-degrading bacterial community in previous studies. A novel anaerobic, thermophilic, and cellulolytic bacterium (strain CSK1) was isolated from MC1. The cellulase activity characteristics of CSK1 were analyzed and evaluated by exploring new measuring conditions via the 3,5-dinitrosalicylic acid (DNS) spectrophotometry and the carboxymethyl cellulose (CMC) saccharogenic power method. The results indicated that the optimal measuring wavelength, reaction temperature, and pH value, were 530 nm, 60°C and 6.0, respectively. The ideal reaction time to achieve stable and significant measuring cellulase activity was about 10 min. Cellulase of CSK1 remained stable when the temperature was below 70°C, and the pH between 5.0 and 10.0, and its activity was quickly reduced when the temperature and pH exceeded such ranges. The cellulase activity of CSK1 reached the highest level on culturing day 8, and high correlations were found among cellulase activity variation, pH and CSK1 biomass change.

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“…The endoglucanase activity was determined by measuring the reducing sugars released after 200 µL of the enzyme was reacted with 200 µL of 2% CMC in 0.05 M phosphate buffer at pH 7.0 (Zhang et al 2009). The mixture was incubated for 30 min at 50 °C, and the reaction was stopped by adding 800 µL of DNS reagent, followed by immersion in boiling water for 5 min.…”

Section: Endoglucanase Assaymentioning

confidence: 99%

Enhanced Endoglucanase Production by Bacillus aerius on Mixed Lignocellulosic Substrates

2016

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Selected carbon sources including soluble carboxymethyl cellulose (CMC), insoluble microcrystalline cellulose (MCC), and single (SS)/mixed lignocellulosic substrates (MS), were evaluated for endoglucanase production by B. aerius S5.2. The lignocellulosic substrates of oil palm empty fruit bunch (EFB), oil palm frond (OPF), rice husk (RH), and their mixture (MS) supported growth of the strain better than CMC and MCC. The maximum endoglucanase activity on MS was 7.3-, 2.6-, 1.7-, and 1.2-fold higher than those recorded on MCC, CMC, EFB/OPF, and RH, respectively. While the reducing sugar concentration of the CMC medium was comparable to that of the EFB and MS media, wide variability was observed in the reducing sugar concentrations among the lignocellulosic substrates. Extremely low levels of sugar were detected in the MCC medium, reflecting its poor digestibility and hence unsuitability for growth and endoglucanase production. Endoglucanase production was predominantly extracellular when the strain was grown on CMC and MS. After seven days of fermentation, there was an approximately 25% reduction in MS dry weight. These findings show that the use of mixed lignocellulosics could potentially reduce the cost of cellulase production. Certain novel aspects of the cellulase system of B. aerius are reported in this study.

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Cellulase Assays

Abstract: Cellulose is a heterogeneous polysaccharide, and its enzymatic hydrolysis requires endoglucanase, exoglucanase (cellobiohydrolase), and beta-glucosidase to work together. We summarize the most commonly used assays for individual enzymes and cellulase mixture.

Cited by 166 publications

References 20 publications

Effects of Pretreatment of Single and Mixed Lignocellulosic Substrates on Production of Endoglucanase by Bacillus aerius S5.2

Effects of Pretreatment of Single and Mixed Lignocellulosic Substrates on Production of Endoglucanase by Bacillus aerius S5.2

Characteristics of cellulase in cellulose-degrading bacterium strain Clostridium straminisolvens (CSK1)

Enhanced Endoglucanase Production by Bacillus aerius on Mixed Lignocellulosic Substrates

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