Mei Jianfeng, Han Yanchao, Zhuang Shihang, Yang Zhikai, Yi Yu, Ying Guoqing
College of Pharmaceutical Science, Zhejiang University of Technology, 18 Chaowang Road, Gongshu District, Hangzhou, 310014, Zhejiang, China.
Bioresour Bioprocess. 2024 Feb 1;11(1):19. doi: 10.1186/s40643-024-00736-w.
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.
胆红素是一种由血红素加氧酶(HO)从血红素水解而来的胆汁色素,在人体中发挥多种功能,包括抗氧化、抗炎和免疫反应抑制活性。胆红素作为临床药物具有很大潜力;然而,目前尚无经济高效的生产方法。因此,本研究探讨了利用表达重组HO的酵母细胞对外源血红素进行生物转化生产胆红素的方法。首先,将编码来自拟南芥的可诱导质体同工酶且去除了质体转运肽序列的血红素加氧酶-1基因(HO1)重组到毕赤酵母GS115细胞中。这导致构建了一种重组毕赤酵母GS115-HO1菌株,其在细胞质中表达活性HO1。之后,对诱导剂甲醇的浓度、诱导培养时间、培养基的pH值以及培养基中提供的山梨醇浓度进行了优化,从而使HO1的产量得到显著提高。随后,将GS115-HO1的全细胞用作催化剂,将氯化血红素(氯高铁血红素)转化为胆红素。结果表明,当向GS115-HO1培养物中加入氯高铁血红素并在30℃下孵育4小时时,胆红素产量为132mg/L。本研究结果为该方法未来用于经济生产胆红素奠定了良好基础。