Production, purification and characterization of an ionic liquid tolerant cellulase from Bacillus sp. isolated from rice paddy field soil

Sriariyanun, Malinee; Tantayotai, Prapakorn; Yasurin, Patchanee; Pornwongthong, Peerapong; Cheenkachorn, Kraipat

doi:10.1016/j.ejbt.2015.11.002

Cited by 63 publications

(44 citation statements)

References 27 publications

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“…Each plate was treated with a UV lamp (254 nm) at a 25-cm distance for 1-30 min and then incubated at 30 C for 10 days. Subsequently, CMC degradation of the colonies that grew on the plates was detected using the Congo Red staining procedure (Sriariyanun et al 2015).…”

Section: Methodsmentioning

confidence: 99%

Production, purification, characterization and usage of a detergent additive of endoglucanase from isolated halotolerant Amycolatopsis cihanbeyliensis mutated strain Mut43

Adıgüzel

Tunçer

2017

Biocatalysis and Biotransformation

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The Amycolatopsis cihanbeyliensis Mut43, which is obtained by UV radiation, exhibited endoglucanase activity of 5.21 U/mL, which was $2.3-fold higher than that of the wild strain (2.04 U/mL). The highest enzyme activity was obtained after 3 days of incubation at 32 C, pH 7.0, 150 rpm, and 6% NaCl in a liquid medium containing 1.5% (w/v) wheat straw (0.25 mm of particle size) and 0.6% (w/v) yeast extract. Enzyme activity was eluted as a single peak (gel filtration chromatography), and Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the corresponding peak revealed a molar mass of 30 kDa. Zymogram analysis confirmed the presence of a single active endoglucanase component. The enzyme was purified to $21-fold, and the mean overall yield was $6%. The purified endoglucanase was active up to 80 C and showed a half-life of 214 min at 60 C in the absence of substrate at pH 8.0. The apparent K m value for the purified endoglucanase was 0.70 mg/mL, while the V max value was 6.20 Units/lg. Endoglucanase activity was reduced (25%) by treatment with 30 U of proteinase K/mg. The addition of Mg þ2 and Ca þ2 (5 mM) enhanced endoglucanase activity. Additionally, endoglucanase activity in the presence of 5 mM SDS or organic solvents was 75 and 50% of maximum activity, respectively. The high levels of enzyme production from A. cihanbeyliensis Mut43 achieved under batch conditions, coupled with the temperature stability, activity over a broad pH range, relatively high stability (70-80%) in the presence of industrial laundry detergents and storage halflives of 45 days at þ4 C and 75 days at À20 C signify the suitability of this enzyme for industrial applications as detergent additive. ARTICLE HISTORY

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Section: Methodsmentioning

confidence: 99%

Production, purification, characterization and usage of a detergent additive of endoglucanase from isolated halotolerant Amycolatopsis cihanbeyliensis mutated strain Mut43

Adıgüzel

Tunçer

2017

Biocatalysis and Biotransformation

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“…100 µl of each microbial consortium from glycerol stocks were inoculated in 20 mL basal medium (containing 1% (w/v) rice straw or cassava pulp, 0.1% NaNo3, 0.1% K2HPO4, 0.1% KCl, 0.05% MgSO4, 0.05% yeast extract, (pH 6.0) [15,16]) or peptone cellulose solution medium (PCS, containing 0.5% (w/v) rice straw or cassava pulp, 0.5% peptone, 0.1% yeast extract, 0.5% NaCl, 0.2% CaCO3, 0.5% filter paper, (pH 8.0) [17]) containing rice straw or cassava pulp as the carbon source. The mixture was incubated at 37°C and 50°C for 3 day in shaker incubator at 200 rpm and kept as a starter culture.…”

Section: Preparation Of Cellulase Enzyme From Microbial Consortiummentioning

confidence: 99%

Analysis of Microbial Consortia with High Cellulolytic Activities for Cassava Pulp Degradation

et al. 2020

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Biogas production is one of the means to manage the cassava pulp waste obtained from the cassava processing plants. The success of the process is determined by the hydrolysis in an anaerobic digester. When the digester failure is found, the new microbial consortium inoculum is introduced to the system with the long period of set up time. This research aimed to construct the endemic microbial consortium by re-cultivating the cellulolytic microbial consortia obtained from cassava pulp and digester wastewater with the expected shorter set up time. Modifications of enrichment and re-cultivation methods by varying the nutrients, pH and temperature improved the enzymatic hydrolysis yields, as reducing sugars, of CMC, rice straw and cassava pulp substrates approximately 9, 3, and 13 times, respectively. To analyze the enzymatic activities of the selected microbial consortia, the cellulase enzyme was extracted, partially purified and analyzed on CMC-zymogram. The ~130 kDa-sized cellulase enzyme was identified with endocellulase activity, and it was considered as a relatively large molecular size molecule compared to most bacterial endocellulases. The selected microbial consortia were tested for their biomass degradation capacities, and the optimal operational condition was obtained at pH 7.0 and 30 °C. This optimal condition showed the proof of the concept that this re-cultivated consortium could be applied in on-site digester with high efficiency.

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“…The kinetic parameters of the purified enzyme revealed that the V max was 1000 μM. min -1 , while K M value was 0.8 mg. mL -1 (Sriariyanun, Tantayotai, Yasurin, Pornwongthong, & Cheenkachorn, 2016). The types and concentration of inhibitors released throughout the pretreatment of lignocellulose depend on the composition of biomass, reaction conditions and pretreatment method.…”

Section: Determination Of Kinetics Parameters Of Tr-cel With Cmc and mentioning

confidence: 99%

Hydrolysis Kinetics of Oil Palm Empty Fruit Bunch in Ionic Liquids and Cellulase Integrated System

Elgharbawy¹,

Alam²,

Moniruzzaman³

et al. 2019

IJC

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Ionic liquids (ILs) are developing as potential solvents in lignocellulose solvation, which enables cellulase accessibility into the substrate. Nevertheless, ILs could result in enzyme deactivation because of the high polarity. Therefore, developing a system of ILs-compatible cellulase (IL-E) to promote lignocellulose conversion into sugars is a challenge in ILs applications. This study used an IL-E to attain high conversion yield of sugars from oil palm empty fruit bunch (EFB). Cellulase (Tr-Cel) from Trichoderma reesei was stable in the ILs, 1-ethyl-3-methyl imidazolium diethyl phosphate [EMIM]DEP and choline acetate [Cho]OAc. The inhibition and deactivation of cellulase were evaluated using the model substrate, carboxymethyl cellulose (CMC) and EFB as a lignocellulosic material to assess the hydrolytic activity. The enzyme kinetics revealed that [Cho]OAc acted as a noncompetitive inhibitor. Additionally, [EMIM]DEP may not be considered as an inhibitor as it increases the Vmax and does not significantly affect the KM. In both cases, the study proved that IL did not result in a severe loss of cellulase activity, which is a promising outcome for one-pot hydrolysis of lignocellulosic materials.

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Production, purification and characterization of an ionic liquid tolerant cellulase from Bacillus sp. isolated from rice paddy field soil

Cited by 63 publications

References 27 publications

Production, purification, characterization and usage of a detergent additive of endoglucanase from isolated halotolerant Amycolatopsis cihanbeyliensis mutated strain Mut43

Production, purification, characterization and usage of a detergent additive of endoglucanase from isolated halotolerant Amycolatopsis cihanbeyliensis mutated strain Mut43

Analysis of Microbial Consortia with High Cellulolytic Activities for Cassava Pulp Degradation

Hydrolysis Kinetics of Oil Palm Empty Fruit Bunch in Ionic Liquids and Cellulase Integrated System

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