Biotransformation of protocatechuic acid

YANG Xiu-Qing1 LIU Ya-Ni1 XI Jing-Wen1

(1.Key Laboratory of Chemical Biology and Molecular Engineering, Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi, China 030006)

【Abstract】[Background] Protocatechuic acid is the main active ingredient of some plants and can be used as a precursor of many polymers as well as various drugs. At present, protocatechuic acid is mainly extracted from plants by chemical methods, with poor extraction efficiency and certain degree of damage to the environment. [Objective] The cloning and heterologous expression of ρ-hydroxybenzoic acid-3-hydroxylase gene ρ-HBA-3H were performed to realize the biotransformation of protocatechuic acid catalyzed by ρ-hydroxybenzoic acid-3-hydroxylase. [Methods] The ρ-hydroxybenzoic acid-3-hydroxylase gene ρ-HBA-3H was amplified by PCR using Rhodococcus sp. R04 genomic DNA as a template and then the recombinant genetic engineering strain BL21 (DE3)/pET21a (+)-ρ-HBA-3H was constructed. The expression of ρ-hydroxybenzoic acid-3-hydroxylase was induced for the biotransformation in the presence of the substrate ρ-hydroxybenzoic acid (ρ-HBA). Besides, the conditions of biotransformation were optimized. [Results] ρ-HBA-3H was highly expressed in E. coli. The yield of protocatechuic acid by biotransformation reached 1.156 g/L. Optimization experiments showed that Mg2+ and Triton X-100 had no effect on the biotransformation efficiency. Increasing the dissolved oxygen content of the reaction system and adding an appropriate amount of Tween-80 promoted the biotransformation. On the basis of continuous cell transformation, a proper amount of glucose can effectively increase the transformation efficiency of engineering bacteria and reduce the consumption of protocatechuic acid. [Conclusion] This study realized the high-efficiency and green production of protocatechuic acid by bio-enzymatic catalysis, and provided a theoretical basis for the industrial production of other important fermentation products.

【Keywords】 Protocatechuic acid; Intermediate metabolites; Biotransformation;

【DOI】

【Funds】 Natural Science Foundation of Shanxi Province (2014011030-3) Natural Science and Coalbed Methane Joint Foundation of Shanxi Province (2015012002) Key Scientific and Technological Project of Shanxi Province (MQ2014-03)

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(Translated by CHEN T)

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This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 46, No. 12, Pages 3378-3387

December 2019

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results
  • 3 Discussion
  • 4 Conclusions
  • References