Characterization and application of a recombinant dopa decarboxylase from Harmonia axyridis for the efficient biosynthesis of dopamine

doi:10.1016/j.cjche.2021.12.006

中国化学工程学报 ›› 2022, Vol. 41 ›› Issue (1): 449-456.DOI: 10.1016/j.cjche.2021.12.006

• Biotechnology and Bioengineering • 上一篇下一篇

Characterization and application of a recombinant dopa decarboxylase from Harmonia axyridis for the efficient biosynthesis of dopamine

Siyuan Gao, Yuanke Guo, Chen Ma, Ding Ma, Kequan Chen, Pingkai Ouyang, Xin Wang

State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China

收稿日期:2021-05-17 修回日期:2021-11-30 出版日期:2022-01-28 发布日期:2022-02-25
通讯作者: Xin Wang,E-mail address:xinwang1988@njtech.edu.cn
基金资助:
This research was funded by the National Natural Science Foundation of China (21576134 and 21706126) and the National Key Research and Development Program (2016YFA0204300)

Characterization and application of a recombinant dopa decarboxylase from Harmonia axyridis for the efficient biosynthesis of dopamine

Siyuan Gao, Yuanke Guo, Chen Ma, Ding Ma, Kequan Chen, Pingkai Ouyang, Xin Wang

State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China

Received:2021-05-17 Revised:2021-11-30 Online:2022-01-28 Published:2022-02-25
Contact: Xin Wang,E-mail address:xinwang1988@njtech.edu.cn
Supported by:
This research was funded by the National Natural Science Foundation of China (21576134 and 21706126) and the National Key Research and Development Program (2016YFA0204300)

摘要/Abstract

摘要： Here, a dopa decarboxylase (DDC) from Harmonia axyridis was heterogeneously expressed in Escherichia coli for the efficient biosynthesis of dopamine. For the production of recombinant DDC, the cultivation conditions including IPTG concentration, temperature and induction time were optimized and obtained an optimal specific enzyme activity of 51.72 U·mg^-1 crude extracts. After the purification of DDC with a recovery yield of 68.79%, its activity was further characterized. The V_max, K_m, K_cat, and K_cat/K_m of DDC for dihydroxyphenylalanine (dopa) were 0.02 mmol·ml^-1·s^-1, 2.328 mmol·ml^-1, 10435.90 s^-1 and 4482.77 ml·mmol^-1·s^-1, respectively. The highest DDC activity was observed at the condition of pH 7.5 and 45 ℃. With the purified DDC, the feasibility to produce dopamine from L-dopa was evaluated. The optimal yield was determined at the following bioconversion conditions: pH of 7.0, the reaction temperature of 40 ℃, 0.4 mmol·L^-1 of PLP and 4 g·L^-1 of L-dopa. Subsequently, a fed-batch process for the production of dopamine was developed and the effect of oxygen was evaluated. The titer, yield and productivity of dopamine reached up to 21.99 g·L^-1, 80.88% and 14.66 g·L^-1·h^-1 at 90 min under anaerobic condition.

关键词: Dopa decarboxylase, Dopamine, Enzyme catalysis, Fed-batch bioconversion

Abstract: Here, a dopa decarboxylase (DDC) from Harmonia axyridis was heterogeneously expressed in Escherichia coli for the efficient biosynthesis of dopamine. For the production of recombinant DDC, the cultivation conditions including IPTG concentration, temperature and induction time were optimized and obtained an optimal specific enzyme activity of 51.72 U·mg^-1 crude extracts. After the purification of DDC with a recovery yield of 68.79%, its activity was further characterized. The V_max, K_m, K_cat, and K_cat/K_m of DDC for dihydroxyphenylalanine (dopa) were 0.02 mmol·ml^-1·s^-1, 2.328 mmol·ml^-1, 10435.90 s^-1 and 4482.77 ml·mmol^-1·s^-1, respectively. The highest DDC activity was observed at the condition of pH 7.5 and 45 ℃. With the purified DDC, the feasibility to produce dopamine from L-dopa was evaluated. The optimal yield was determined at the following bioconversion conditions: pH of 7.0, the reaction temperature of 40 ℃, 0.4 mmol·L^-1 of PLP and 4 g·L^-1 of L-dopa. Subsequently, a fed-batch process for the production of dopamine was developed and the effect of oxygen was evaluated. The titer, yield and productivity of dopamine reached up to 21.99 g·L^-1, 80.88% and 14.66 g·L^-1·h^-1 at 90 min under anaerobic condition.

Key words: Dopa decarboxylase, Dopamine, Enzyme catalysis, Fed-batch bioconversion

Siyuan Gao, Yuanke Guo, Chen Ma, Ding Ma, Kequan Chen, Pingkai Ouyang, Xin Wang. Characterization and application of a recombinant dopa decarboxylase from Harmonia axyridis for the efficient biosynthesis of dopamine[J]. 中国化学工程学报, 2022, 41(1): 449-456.

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Characterization and application of a recombinant dopa decarboxylase from Harmonia axyridis for the efficient biosynthesis of dopamine

Characterization and application of a recombinant dopa decarboxylase from Harmonia axyridis for the efficient biosynthesis of dopamine

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