Recombinant Horseradish Peroxidase C1A Immobilized on Hydrogel Matrix for Dye Decolorization and Its Mechanism on Acid B
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Recombinant Horseradish Peroxidase C1A Immobilized on Hydrogel Matrix for Dye Decolorization and Its Mechanism on Acid Blue 129 Decolorization Ya-Jing Wang 1 & Kai-Zhong Xu 1 & Hui Ma 1 & Xiang-Ru Liao 1 & Guang Guo 2 & Fang Tian 2 & Zheng-Bing Guan 1 Received: 21 January 2020 / Accepted: 23 June 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
In this study, horseradish peroxidase C1A (HRP C1A) from Armoracia rusticana was expressed in Escherichia coli as an inclusion body. Subsequently, an active recombinant HRP C1A was obtained by refolding gradually using dilution-ultrafiltration. The recombinant HRP C1A was immobilized on agarose-chitosan hydrogel at 86.9 ± 2.5% of immobilization efficiency. After immobilization of the recombinant HRP C1A, the pH and temperature stability were improved and the reusability of the recombinant HPR C1A was achieved. The immobilized HRP C1A activity was retained above 80% after 6 cycles. The immobilized recombinant HRP C1A was used for the decolorization of four various dyes, including acid blue 129 (AB129), methyl blue (MB), methyl red (MR), and trypan blue (TB). The decolorization rates are all more than 70%, among which the decolorization effect of AB129 was the most significant (the decolorization rate was 76.3 ± 1.6%). Furthermore, a plausible decolorization pathway for AB129 was proposed based on the identified intermediates by ultraperformance liquid chromatography coupled with mass spectrometry (UPLC-MS). This is the first report of the putative mechanism on the decolorization of AB129 by HRP. Keywords Heterologous expression . Refold . Immobilization . Horseradish peroxidase C1A . Dye decolorization . Acid blue 129
* Zheng-Bing Guan [email protected]
1
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, People’s Republic of China
2
College of Environmental Engineering, Nanjing Institute of Technology, Nanjing 211167, People’s Republic of China
Applied Biochemistry and Biotechnology
Introduction Synthetic dyes are widely used in the textile industry. At present, the variety of dyes increases rapidly and develops in the direction of anti-photodegradation and anti-oxidation, making the treatment of dye wastewater more difficult. This textile wastewater poses a potential threat to the Earth’s environment and ecosystems. According to the chemical structure of the chromophore, dyes are classified into azo, anthraquinone, triphenylmethane, heterocyclic, and polymeric dyes [1]. These dyes are carcinogenic and mutagenic, which must be treated before discharge [2, 3]. Traditional decolorization methods of textile wastewater include physical and chemical methods (such as electrochemical method and oxidation method) [4]. However, they are generally high energy consumption, high processing cost, and easy secondary pollution, which greatly limits the wide application [5–8]. Conversely, the bioenzymatic method has the advantages of mild reaction conditions, high
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