Production of bilirubin by biotransformation of biliverdin using recombinant Escherichia coli cells

Mei, Jianfeng; Wu, Xia; Zheng, Sujing; Yi, Yu; Wang, Xudong; Ying, Guoqing

doi:10.1007/s00449-021-02679-4

Cited by 2 publications

(1 citation statement)

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“…The assay for measuring HO1 activity was as follows: 50 μL of 0.5 mmol/L hemin solution (dissolved in 0.05 mol/L Na 2 CO 3 ), 449 μL of 0.1 mol/L pH 7.0 sodium citrate buffer, 40 μL of 10 g/L bovine serum albumen (BSA), 50 μL of 40 mmol/L d -glucose-6-phosphate disodium, 1 μL of 1 U/μL glucose-6-phosphate dehydrogenase solution, 10 μL of 20 mmol/L NADPH, 200 μL of the crude biliverdin reductase preparation, which was prepared according to the method described in the reference (Mei et al 2022 ), and 200 μL of GS115-HO1 cell lysate were mixed in a 1.5-mL centrifuge tube and incubated for 15 min at 30 °C. The A 450 of the enzyme reaction mixture was measured, and the concentration of bilirubin in it was calculated using the standard curve for bilirubin concentration—A 450 .…”

Section: Methodsmentioning

confidence: 99%

Production of biliverdin by biotransformation of exogenous heme using recombinant Pichia pastoris cells

Mei,

Han,

Zhuang

et al. 2024

Bioresour. Bioprocess.

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Biliverdin, a bile pigment hydrolyzed from heme by heme oxygenase (HO), serves multiple functions in the human body, including antioxidant, anti-inflammatory, and immune response inhibitory activities. Biliverdin has great potential as a clinical drug; however, no economic and efficient production method is available currently. Therefore, the production of biliverdin by the biotransformation of exogenous heme using recombinant HO-expressing yeast cells was studied in this research. First, the heme oxygenase-1 gene (HO1) encoding the inducible plastidic isozyme from Arabidopsis thaliana, with the plastid transport peptide sequence removed, was recombined into Pichia pastoris GS115 cells. This resulted in the construction of a recombinant P. pastoris GS115-HO1 strain that expressed active HO1 in the cytoplasm. After that, the concentration of the inducer methanol, the induction culture time, the pH of the medium, and the concentration of sorbitol supplied in the medium were optimized, resulting in a significant improvement in the yield of HO1. Subsequently, the whole cells of GS115-HO1 were employed as catalysts to convert heme chloride (hemin) into biliverdin. The results showed that the yield of biliverdin was 132 mg/L when hemin was added to the culture of GS115-HO1 and incubated for 4 h at 30 °C. The findings of this study have laid a good foundation for future applications of this method for the economical production of biliverdin. Graphical Abstract

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Section: Methodsmentioning

confidence: 99%