Immobilization and characterization of tannase from Penicillium rolfsii CCMB 714 and its efficiency in apple juice clari
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ORIGINAL PAPER
Immobilization and characterization of tannase from Penicillium rolfsii CCMB 714 and its efficiency in apple juice clarification Priscilla Macedo Lima Andrade1,2 · Luciana Baptista1 · Camila Oliveira Bezerra1 · Rosane Marina Peralta3 · Aristóteles Góes‑Neto4 · Ana Paula Trovatti Uetanabaro1 · Andréa Miura da Costa1 Received: 28 April 2020 / Accepted: 12 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract In the present study a newly tannase enzyme from Penicillium rolfsii CCMB 714 was immobilized on calcium alginate and its efficiency in clarifying juice was analyzed. The efficiency of immobilization was 99.5% and the enzyme maintained an activity above 50% after 6 cycles of reuse. Immobilization of tannase enzyme changed the optimum temperature from 33 to 44 °C, and the optimum pH from 4.0 to 4.3. The immobilized tannase enzyme showed greater thermal and pH stability when compared to the free form, maintaining 80% of its activity at 50 °C and more than 50% of residual activity at all pH tested for up to 16 h. The immobilized enzyme was also resistant to almost all additives used (cations, SDS and EDTA). When tested in apple juice, the immobilized tannase efficiently clarified the juice showing the reduction of more than 70% in turbidity. Such results showing that the P. rolfsii CCMB 714 immobilized tannase was suitable to clarify apple juice and have a potential for future beverage industrial application mainly in juice processing. Keywords Enzyme immobilization · Juice clarification · Tannase · Fungi · Submerged culture · Enzyme reuse
Introduction Tannase or tannin acylhydrolase (EC 3.1.1.20) catalyzes the hydrolysis of ester linkages present in hydrolyzable tannins and esters of gallic acid [1]. Hydrolyzable tannins are found in a variety of plant tissues, they have high molecular weight and are composed of glucose and gallic acid molecules [2]. Tannases can be found among the plant, animal and microbial kingdoms. In terms of filamentous fungi, the Aspergillus and Penicillium genera have been described as excellent
* Andréa Miura da Costa [email protected] 1
Departament of Biological Sciences, UESC - Universidade Estadual de Santa Cruz, Jorge Amado Highway, Km 16, Ilhéus, BA 45662‑900, Brazil
2
IFBA - Instituto Federal da Bahia, Rodovia BR 367, Km 57.5, Fontana I, Porto Seguro, BA, Brazil
3
Departament of Biochemistry, UEM - Universidade Estadual de Maringá, Maringá, PR, Brazil
4
Biological Science Institute, UFMG – Universidade Federal de Minas Gerais, Antônio Carlos 6627 Avenue, Belo Horizonte, MG, Brazil
tannase producers in both solid state and submerged cultures [3–6]. Tannases present a wide range of commercial applications in food, beverage, chemical, pharmaceutical, and animal feed industries [5]. However, as the presence of high amounts of tannins in some fruits can be responsible for the formation of turbidity and sediments, as well as color, bitterness, and astringency of the juice during its storage, the clarification of
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