Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins
Background: Theabrownins (TBs), which are produced from tea leaves, have been shown to have biological. To extract biologically active chemicals from polyphenol structures, an oxidizing enzyme (i.e. laccase) is used to oxidize the polyphenol structure, releasing smaller and more soluble molecular co...
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2023-01-01
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| Series: | Biomedical and Biotechnology Research Journal |
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| Online Access: | http://www.bmbtrj.org/article.asp?issn=2588-9834;year=2023;volume=7;issue=4;spage=633;epage=640;aulast=Vu |
| _version_ | 1827270306995109888 |
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| author | Dinh Giap Vu Nhat Minh Dang Phuong Dai Nguyen Nguyen Thi Dung Ha Vinh The Mac |
| author_facet | Dinh Giap Vu Nhat Minh Dang Phuong Dai Nguyen Nguyen Thi Dung Ha Vinh The Mac |
| author_sort | Dinh Giap Vu |
| collection | DOAJ |
| description | Background: Theabrownins (TBs), which are produced from tea leaves, have been shown to have biological. To extract biologically active chemicals from polyphenol structures, an oxidizing enzyme (i.e. laccase) is used to oxidize the polyphenol structure, releasing smaller and more soluble molecular compounds, including TBs. This is a biologically active compound that is beneficial to human health. Therefore, using laccase to purify is significant in enhancing the oxidation process of polyphenols from fresh tea leaves to improve the efficiency of harvesting theabrownins. Methods: to purify laccase, we use a diethylaminoethyl-cellulose (DEAE), Sephadex G-75, and Q Sepharose® columns in a three-step column chromatography procedure. In addition, TBs formed by hydrolysis were determined by high-performance liquid chromatography equipped with a refractive index detector (ultra violet-Vis 271 nm) and ICSep WA-1 Analysis column. Results: In this study, CliLac was effectively purified with a specific activity of 7.58 U/mg from strain Clitopilus prunulus BV18. Using DEAE, Sephadex G-75, and Q Sepharose® columns in a three-step column chromatography procedure, the enzyme was purified to a level of 21.4-fold purity. The enzyme with a molecular weight of 55.2 kDa demonstrated increased pH stability in the acidic range. Biochemical properties of CliLac showed that the optimum pH and temperature were 5.0 and 50°C, respectively. The activity retention was over 80% at pH 5.0 for more than 7 h of incubation. CliLac denatured at temperatures over 60°C. TBs production release increased up to 159 ppm after 72 h of incubation using a purified CliLac (10 U/gds) at 50°C. Conclusions: The study indicates that CliLac is appropriate for oxidizing the polyphenol structure and releases the TBs, which are smaller and more soluble molecular compounds. |
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| format | Article |
| id | doaj.art-76d63013297541e183f19fbc883d919d |
| institution | Directory Open Access Journal |
| issn | 2588-9834 2588-9842 |
| language | English |
| last_indexed | 2025-03-22T05:19:20Z |
| publishDate | 2023-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Biomedical and Biotechnology Research Journal |
| spelling | doaj.art-76d63013297541e183f19fbc883d919d2024-04-27T04:17:45ZengWolters Kluwer Medknow PublicationsBiomedical and Biotechnology Research Journal2588-98342588-98422023-01-017463364010.4103/bbrj.bbrj_266_23Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrowninsDinh Giap VuNhat Minh DangPhuong Dai Nguyen NguyenThi Dung HaVinh The MacBackground: Theabrownins (TBs), which are produced from tea leaves, have been shown to have biological. To extract biologically active chemicals from polyphenol structures, an oxidizing enzyme (i.e. laccase) is used to oxidize the polyphenol structure, releasing smaller and more soluble molecular compounds, including TBs. This is a biologically active compound that is beneficial to human health. Therefore, using laccase to purify is significant in enhancing the oxidation process of polyphenols from fresh tea leaves to improve the efficiency of harvesting theabrownins. Methods: to purify laccase, we use a diethylaminoethyl-cellulose (DEAE), Sephadex G-75, and Q Sepharose® columns in a three-step column chromatography procedure. In addition, TBs formed by hydrolysis were determined by high-performance liquid chromatography equipped with a refractive index detector (ultra violet-Vis 271 nm) and ICSep WA-1 Analysis column. Results: In this study, CliLac was effectively purified with a specific activity of 7.58 U/mg from strain Clitopilus prunulus BV18. Using DEAE, Sephadex G-75, and Q Sepharose® columns in a three-step column chromatography procedure, the enzyme was purified to a level of 21.4-fold purity. The enzyme with a molecular weight of 55.2 kDa demonstrated increased pH stability in the acidic range. Biochemical properties of CliLac showed that the optimum pH and temperature were 5.0 and 50°C, respectively. The activity retention was over 80% at pH 5.0 for more than 7 h of incubation. CliLac denatured at temperatures over 60°C. TBs production release increased up to 159 ppm after 72 h of incubation using a purified CliLac (10 U/gds) at 50°C. Conclusions: The study indicates that CliLac is appropriate for oxidizing the polyphenol structure and releases the TBs, which are smaller and more soluble molecular compounds.http://www.bmbtrj.org/article.asp?issn=2588-9834;year=2023;volume=7;issue=4;spage=633;epage=640;aulast=Vuabrowninsbasidiomycetesclitopilus prunuluslaccasetea polyphenols |
| spellingShingle | Dinh Giap Vu Nhat Minh Dang Phuong Dai Nguyen Nguyen Thi Dung Ha Vinh The Mac Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins Biomedical and Biotechnology Research Journal abrownins basidiomycetes clitopilus prunulus laccase tea polyphenols |
| title | Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins |
| title_full | Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins |
| title_fullStr | Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins |
| title_full_unstemmed | Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins |
| title_short | Purification of laccase from the fungus Clitopilus prunulus BV18 and its application in bioconversion of tea polyphenols to bioactive theabrownins |
| title_sort | purification of laccase from the fungus clitopilus prunulus bv18 and its application in bioconversion of tea polyphenols to bioactive theabrownins |
| topic | abrownins basidiomycetes clitopilus prunulus laccase tea polyphenols |
| url | http://www.bmbtrj.org/article.asp?issn=2588-9834;year=2023;volume=7;issue=4;spage=633;epage=640;aulast=Vu |
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