Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris

Sriyapai, Thayat; Somyoonsap, Peechapack; Matsui, Kenji; Kawai, Fusako; Chansiri, Kosum

doi:10.1016/j.jbiosc.2010.12.024

Cited by 70 publications

(36 citation statements)

References 28 publications

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“…These observations presumably suggested that the xylanase from Actinomadura sp. Cpt20 was a monomeric protein comparable to those previously reported for other xylanases [13,37,38].…”

Section: Identification and Molecular Phylogeny Of The Microorganismsupporting

confidence: 83%

“…S14 and Actinomadura sp. FC7 strains, which were stable over a wide pH range [13][14][15]. Therefore, the tolerance to a broad pH range, especially to alkaline and high-temperature conditions, exhibited by the xylanase from Actinomadura sp.…”

Section: Effect Of Ph On Xylanase Activity and Stabilitymentioning

confidence: 99%

“…Various xylan-degrading thermotolerant actinomycetes, such as Actinomadura [13][14][15], Nonomuraea [16], Microtetraspora [17], Streptomyces [18,19], and Thermomonospora [20,21], have been purified and characterized. Some research seem, however, to have paid special attention to xylanases from actinomycete origins particularly because of the promising potential they offer for a wide range of biotechnological applications particularly in the pulp and paper industry [22].…”

Section: Introductionmentioning

confidence: 99%

“…A few studies report on the purification and characterization of xylanases from Actinomadura sp. [13][14][15]. This is due to the limited potential for industrial application, since the xylanases previously purified from Actinomadura species had poor enzyme stability and catalytic activity.…”

Section: Introductionmentioning

confidence: 99%

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Purification and Biochemical Characterization of a Highly Thermostable Xylanase from Actinomadura sp. Strain Cpt20 Isolated from Poultry Compost

Taibi

Saoudi

Boudelaa

et al. 2011

Appl Biochem Biotechnol

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An extracellular thermostable xylanase from a newly isolated thermophilic Actinomadura sp. strain Cpt20 was purified and characterized. Based on matrix-assisted laser desorption-ionization time-of-flight mass spectrometry analysis, the purified enzyme is a monomer with a molecular mass of 20,110.13 Da. The 19 residue N-terminal sequence of the enzyme showed 84% homology with those of actinomycete endoxylanases. The optimum pH and temperature values for xylanase activity were pH 10 and 80 °C, respectively. This xylanase was stable within a pH range of 5-10 and up to a temperature of 90 °C. It showed high thermostability at 60 °C for 5 days and half-life times at 90 °C and 100 °C were 2 and 1 h, respectively. The xylanase was specific for xylans, showing higher specific activity on soluble oat-spelt xylan followed by beechwood xylan. This enzyme obeyed the Michaelis-Menten kinetics, with the K (m) and k (cat) values being 1.55 mg soluble oat-spelt xylan/ml and 388 min(-1), respectively. While the xylanase from Actinomadura sp. Cpt20 was activated by Mn(2+), Ca(2+), and Cu(2+), it was, strongly inhibited by Hg(2+), Zn(2+), and Ba(2+). These properties make this enzyme a potential candidate for future use in biotechnological applications particularly in the pulp and paper industry.

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“…These observations presumably suggested that the xylanase from Actinomadura sp. Cpt20 was a monomeric protein comparable to those previously reported for other xylanases [13,37,38].…”

Section: Identification and Molecular Phylogeny Of The Microorganismsupporting

confidence: 83%

Section: Effect Of Ph On Xylanase Activity and Stabilitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Purification and Biochemical Characterization of a Highly Thermostable Xylanase from Actinomadura sp. Strain Cpt20 Isolated from Poultry Compost

Taibi

Saoudi

Boudelaa

et al. 2011

Appl Biochem Biotechnol

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“…S27 (Li et al, 2009) and Actinomadura sp. S14 (Sriyapai, 2011). Thermophilic bacteria can survive at high temperatures 50-65 °C (Soeka et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

CONDITION OPTIMIZATION PRODUCTION XYLANASE THERMOSTABLE BY Bacillus licheniformis TS10 USING SUBSTRATE OIL PALM EMPTY FRUIT BUNCHES

Safitri¹,

Muharni²,

Kusumawati³

2017

BIOV

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ABSTRAKOptimasi produksi xilanase termostabil oleh Bacillus licheniformis TS10 menggunakan substrat tandan kosong kelapa sawit telah dilakukan di Laboratorium Genetika dan Bioteknologi, Jurusan Biologi Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Sriwijaya, Indralaya. Penelitian dilakukan sejak November 2015 hingga Januari 2016. Penelitian ini bertujuan untuk mengetahui potensi tandan kosong kelapa sawit (TKKS) sebagai substrat untuk produksi xilanase termostabil oleh Bacillus licheniformis TS10 serta kondisi optimum suhu, pH dan konsentrasi substrat pada proses fermentasi. Metode penelitian adalah dengan membuat kurva pertumbuhan dan kurva produksi enzim dari Bacillus licheniformis TS10 pada substrat TKKS. Jumlah sel bakteri ditentukan dengan menggunakan spektrophotometer UV-VIS dan total plate count (TPC) melalui kurva standar. Optimasi dilakukan pada variasi pH (5, 6, 7, 8, 9), suhu (50°C, 60 °C, 70 °C, 80 °C) dan substrat (1%, 2%, 3%, 4%). Uji aktivitas pada masingmasing pH, suhu dan substrat menggunakan metode DNS dengan mengukur aktivitas enzim berdasarkan hasil gula reduksi yang dilepaskan oleh substrat dengan menggunakan dinitrosalicylic acid (DNS). Berdasarkan hasil penelitian dapat diketahui bahwa tandan kosong kelapa sawit (TKKS) memiliki potensi sebagai substrat untuk produksi xilanase termostabil oleh Bacillus licheniformis TS10, produksi xilanase termostabil dari Bacillus licheniformis TS10 pada substrat TKKS memiliki kondisi optimum pada pH 6, suhu 80 °C dan konsentrasi substrat sebesar 4%.Kata kunci : Optimasi, Xilanase Termostabil, Bacillus licheniformis TS10, Tandan Kosong Kelapa Sawit. ABSTRACTOptimization of thermostable xylanase production by Bacillus licheniformis TS10 using substrate oil palm empty fruit bunches has been conducted from November 2015 to January 2016 in the Laboratory of Genetics and Biotechnology, Department of Biology, Faculty of Mathematics and Natural Sciences, University of Sriwijaya, Indralaya. The aims to determine the potential of oil palm empty fruit bunches (EFB) as a substrate for the production of thermostable xylanase by Bacillus licheniformis TS10 and the optimum conditions of temperature, pH and substrate concentration in the fermentation process. The research method are to make the growth curve and the curve of Bacillus licheniformis TS10 enzyme production on EFB substrate. The number of bacterial cells was determined using a UV-VIS spectrophotometer and total plate count (TPC) through a standard curve. The optimization performed of pH (5, 6, 7, 8, 9), temperature (50 °C, 60 °C, 70 °C, 80 °C) and substrate (1%, 2%, 3%, 4%). The test activity in various pH, temperature and substrate using methods DNS by measuring enzyme activity based on the reducing sugar released by the substrate by using Dinitro Salicylic Acid (DNS). The results showed that oil palm empty fruit bunches (EFB) has a potential as a substrate for the production of thermostable xylanase by Bacillus licheniformis TS10, thermostable xylanase production of Bacillus licheniformis TS1...

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Chaotropic heat treatment resolves native‐like aggregation of a heterologously produced hyperthermostable laminarinase

Westphal

Geerke-Volmer

Mierlo

et al. 2017

Biotechnology Journal

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Production of hyperthermostable enzymes in mesophilic hosts frequently causes undesired aggregation of these proteins. During production of Pyrococcus furiosus endo-β-1,3 glucanase (LamA) in Escherichia coli, soluble and insoluble species form. Here, the authors address the composition of this mixture, including the nature of LamA conformers, and establish a method to increase the yield of native monomer. With gel electrophoresis, size-exclusion chromatography, light scattering, circular dichroism and enzyme kinetics the authors show that approximately 50 % of heterologously produced LamA is soluble, and that 40 % of this fraction constitutes native-like oligomers and non-native monomers. Soluble oligomers display, like native LamA monomer, substrate inhibition, although with poor activity. Treatment of soluble oligomers with 3 M guanidinium hydrochloride at 80 °C yields up to 75 % properly active monomer. Non-native monomer shows low specific activity without substrate inhibition. Incubating non-native monomer with 3 M guanidinium hydrochloride at 80 °C causes formation of 25 % native LamA. Also, a large amount of insoluble LamA aggregates can be converted into soluble native monomer by application of this procedure. Thus, chaotropic heat treatment can improve the yield and quality of hyperthermostable proteins that form aberrant species during production in E. coli.

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Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris

Cited by 70 publications

References 28 publications

Purification and Biochemical Characterization of a Highly Thermostable Xylanase from Actinomadura sp. Strain Cpt20 Isolated from Poultry Compost

Purification and Biochemical Characterization of a Highly Thermostable Xylanase from Actinomadura sp. Strain Cpt20 Isolated from Poultry Compost

CONDITION OPTIMIZATION PRODUCTION XYLANASE THERMOSTABLE BY Bacillus licheniformis TS10 USING SUBSTRATE OIL PALM EMPTY FRUIT BUNCHES

Chaotropic heat treatment resolves native‐like aggregation of a heterologously produced hyperthermostable laminarinase

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