[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

要旨

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.

材料

製品番号

ブランド

製品内容

E7023

Sigma-Aldrich

エチルアルコール（純粋）, Molecular Biology

09-0770

Sigma-Aldrich

エタノール, JIS special grade, ≥99.5%

362808

Sigma-Aldrich

Reagent Alcohol, reagent grade

09-0780

Sigma-Aldrich

エタノール, SAJ first grade, ≥99.5%

493511

Sigma-Aldrich

エチルアルコール（純粋）, 190 proof, ACS spectrophotometric grade, 95.0%

02857

Sigma-Aldrich

エタノール, purum, fine spirit, denaturated with 4.8% methanol, F25 METHYL¹, ~96% (based on denaturant-free substance)

E-029

Supelco

エタノール-50, 50 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-040

Supelco

エタノール-10, 10 mg/dL in H₂O, pack of 10 × 1.2 mL ampules, certified reference material, Cerilliant^®

48075

Supelco

エタノール溶液, certified reference material, 2000 μg/mL in methanol

32221

Sigma-Aldrich

エタノール, puriss. p.a., absolute, ≥99.8% (GC)

E-030

Supelco

エタノール-80, 80 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-053

Supelco

エタノール-500, 500 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-031

Supelco

エタノール-100 (10アンプル／キット), 100 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-033

Supelco

エタノール-300, 300 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E2385

Supelco

エタノール標準品10% (v/v), 10 % (v/v) in H₂O, analytical standard

E-041

Supelco

エタノール-150, 150 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

09-0851

Sigma-Aldrich

エタノール, ≥99.5%, suitable for HPLC

E-032

Supelco

エタノール-200（10アンプル／キット）, 200 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

09-0790

Sigma-Aldrich

エタノール, JIS special grade, 94.8-95.8%

09-0852

Sigma-Aldrich

エタノール, ≥99.5%

E-056

Supelco

エタノール-20（10アンプル／キット）, 20 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-035

Supelco

エタノール溶液, 25 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-036

Supelco

エタノール-400（10アンプル／キット）, 400 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-034

Supelco

Ethanol Calibration Kit, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

187380

Sigma-Aldrich

エチルアルコール（変性）, denatured, reagent grade

09-0850

Sigma-Aldrich

エタノール, ≥99.5%, suitable for absorption spectrum analysis

2858

Sigma-Aldrich

Ethanol Fixative 80% v/v, suitable for fixing solution (blood films)

E-045

Supelco

エタノール溶液, 40 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

09-0800

Sigma-Aldrich

エタノール, 94.8-95.8%

676829

Sigma-Aldrich

Reagent Alcohol, anhydrous, ≤0.005% water