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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Environmental Science / Faculty of Environmental Earth Science >
Peer-reviewed Journal Articles, etc >
Gene cloning and characterization of a vanadium-dependent bromoperoxidase from the red alga Laurencia saitoi, a producer of brominated diterpenoids and triterpenoids
| Title: | Gene cloning and characterization of a vanadium-dependent bromoperoxidase from the red alga Laurencia saitoi, a producer of brominated diterpenoids and triterpenoids |
| Authors: | Kaneko, Kensuke Browse this author | | Kobayashi, Daiki Browse this author | | Masaki, Shiro Browse this author | | Washio, Kenji Browse this author →KAKEN DB | | Morikawa, Masaaki Browse this author →KAKEN DB | | Okino, Tatsufumi Browse this author →KAKEN DB |
| Keywords: | Laurencia saitoi | | Vanadium-dependent bromoperoxidase (V-BPO) | | Bromination activity | | Rhodophyta |
| Issue Date: | 10-Apr-2023 |
| Publisher: | Springer |
| Journal Title: | Journal of Applied Phycology |
| Publisher DOI: | 10.1007/s10811-023-02953-w |
| Abstract: | The gene encoding vanadium-dependent bromoperoxidase (V-BPO) was cloned for the first time from the red alga Laurencia saitoi, which produces pharmaceutically promising brominated diterpenoids and triterpenoids. The molecular weight of V-BPO from L. saitoi (LsVBPO1) was the highest (77.0 kDa) among previously reported V-BPOs from Laurencia with a peptide insertion Asn194-Ser221 containing short Gln repeats. It shares approximately 60% amino acid sequence identity with V-BPOs from L. nipponica (LnVBPO1 and LnVBPO2) and L. okamurae (LoVBPO1a and LoVBPO2a). Heterologously expressed LsVBPO1 in Escherichia coli was partially purified and exhibited low but significant bromination activity of 38 U mg(-1) protein using monochlorodimedone. The pH optimum was 8.0, which was more alkaline than that for LnVBPOs and LoVBPO2a (pH 7.0). The K-m for H2O2 was 0.04 mM, comparable to LnVBPO1 (0.026 mM), LnVBPO2 (0.025 mM), and LoVBPO2a (0.014 mM). LsVPBO1 retained its bromination activity until 45 degrees C for 20 min. When incubated at 55 degrees C for 20 min, catalytic activity decreased rapidly, as shown for LnVBPO1 and LoVBPO2a (retained at 45 degrees C, decreased at 55 degrees C) and LnVBPO2 (retained at 55 degrees C, decreased at 65 degrees C). Unlike other V-BPOs from Laurencia (LnVBPO1, LnVBPO2, and LoVBPO2a), dialysis and concentration during purification process were rapidly inactivated LsVBPO1, suggesting its structural instability. |
| Rights: | This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s10811-023-02953-w |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/91590 |
| Appears in Collections: | 環境科学院・地球環境科学研究院 (Graduate School of Environmental Science / Faculty of Environmental Earth Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 沖野 龍文
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