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  • [Cloning, expression and characterization of alcohol dehydrogenase and aldehyde dehydrogenase from Bacillus pseudofirmus OF4].

[Cloning, expression and characterization of alcohol dehydrogenase and aldehyde dehydrogenase from Bacillus pseudofirmus OF4].

Wei sheng wu xue bao = Acta microbiologica Sinica (2013-07-19)
Jiansong Ju, Ning Ma, Ranran Zhao, Jingwei Liu, Shujing Xu, Baohua Zhao
PMID23858711
ABSTRACT

To clone and characterize the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) from Bacillus pseudofirmus OF4. Genes of adh and aldh were cloned by PCR; expression vectors pET-Ahd and pET-Aldh were constructed and expressed in Escherichia coli BL21 (DE3). After Ni-NTA column chromatography purification, the protein was characterized. The optimal temperature and pH of ALDH was 35 degrees C and 8.0, the specific activities of ALDH was 979.6 U/mg protein, the thermostability at 25 degrees C and 35 degrees C was better than at 45 degrees C. Although the expression level of ADH was too low to purify, but it was found that ADH had high catalytic activities by experiments of co-expression and ethanol tolerance. Adh and aldh from B. pseudofirmus OF4 were cloned successfully. Co-expression of double genes could greatly increase the host strain on ethanol tolerance.

MATERIALI
N° Catalogo
Marchio
Descrizione del prodotto
459844
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, ACS reagent, ≥99.5%
493546
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Ethyl alcohol, Pure, 200 proof, meets USP testing specifications
E7148
Sigma-Aldrich
Ethyl alcohol, Pure, 190 proof, for molecular biology
51976
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Etanolo, BioUltra, for molecular biology, ≥99.8%, (absolute alcohol, without additive, A15 o1)
02483
Supelco
Etanolo, standard for GC
02857
Sigma-Aldrich
Etanolo, purum, fine spirit, denaturated with 4.8% methanol, F25 METHYL1, ~96% (based on denaturant-free substance)
48075
Supelco
Ethanol solution, certified reference material, 2000 μg/mL in methanol
E-029
Supelco
Ethanol-50, 50 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
E-031
Supelco
Ethanol-100 (10 ampules/kit), 100 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
PHR1070
Supelco
Etanolo, Pharmaceutical Secondary Standard; Certified Reference Material
E-040
Supelco
Ethanol-10, 10 mg/dL in H2O, pack of 10 × 1.2 mL ampules, certified reference material, Cerilliant®
02851
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Etanolo, for residue analysis
E-030
Supelco
Ethanol-80, 80 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
277649
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Reagent Alcohol, anhydrous, ≤0.003% water
E-053
Supelco
Ethanol-500, 500 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
E-033
Supelco
Ethanol-300, 300 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
02882
Sigma-Aldrich
Etanolo, purum, absolute ethanol, denaturated with 2% 2-butanone, A15 MEK1, ≥99.8% (based on denaturant-free substance)
02890
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Etanolo, purum, fine spirit, denaturated with 2% 2-butanone, F25 MEK1, ~96% (based on denaturant-free substance)
E-032
Supelco
Ethanol-200 (10 ampules/kit), 200 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
E-041
Supelco
Ethanol-150, 150 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
676829
Sigma-Aldrich
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E-034
Supelco
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E-035
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Ethanol-25, 25 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
E-056
Supelco
Ethanol-20 (10 ampules/kit), 20 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
E-036
Supelco
Ethanol-400 (10 ampules/kit), 400 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®
29221
Sigma-Aldrich
Etanolo, tested according to Ph. Eur.
02891
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Etanolo, purum, secunda spirit, denaturated with 2% 2-butanone, S15, ~96% (based on denaturant-free substance)
E-045
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Ethanol-40, 40 mg/dL in H2O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant®