Gene cloning, phenol-responsive transcriptional profiling and recombinant protein characterization of phenol hydroxylase in Candida tropicalis GY8.

Authors

Jianhua Lv, Xiangnan Gu, Minmin Hui, Yingjie Luo, Wenbo Luo, Huiting Zhao, Shuqin Yin, Yan Feng, Zhiquan Xue

Published in

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]. Jun 02, 2025. Epub Jun 02, 2025.

Abstract

Phenol hydroxylase (PH) plays a central role in phenol biodegradation pathways. In this study, the enzymatic activity of PH in Candida tropicalis GY8 was evaluated following phenol-induced cultivation, and oxidation products were analyzed via high-performance liquid chromatography (HPLC). Two phenol hydroxylase genes, CtPHE1 and CtPHE2, were identified, and their expression patterns were examined using RT-qPCR. Functional analysis of the corresponding proteins was conducted by cloning and expressing the genes in Escherichia coli BL21(DE3). Enzymatic activity reached its maximum after 15 h of cultivation (p < 0.01), and phenol conversion to catechol was verified by HPLC. RT-qPCR results indicated peak transcript levels at 10 h, with CtPHE2 showing consistently higher expression than CtPHE1 at 5, 10, and 15 h (p < 0.01). Both recombinant proteins were capable of phenol degradation, with CtPHE2 exhibiting greater catalytic activity. These findings provide essential molecular information for advancing the understanding of phenol biodegradation mechanisms.

PMID:
40455391
Bibliographic data and abstract were imported from PubMed on 02 Jun 2025.

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