A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells
Abstract Background Triterpenoid compounds such as betulinic acid (BA) and oleanolic acid (OA) exhibit considerable pharmacological activities. However, their current production primarily relies on plant extraction and chemical synthesis, methods that are often plagued by low efficiency, complex ext...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-06-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-025-02733-w |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850224153375801344 |
|---|---|
| author | Luran Geng Jiale Cui Changyixin Xiao Linlin Xu Fengjiao Yue Ting Zhang Mengran Zhan Zichang Lu Yuzhe Ren Chunsheng Wang Jing Yin |
| author_facet | Luran Geng Jiale Cui Changyixin Xiao Linlin Xu Fengjiao Yue Ting Zhang Mengran Zhan Zichang Lu Yuzhe Ren Chunsheng Wang Jing Yin |
| author_sort | Luran Geng |
| collection | DOAJ |
| description | Abstract Background Triterpenoid compounds such as betulinic acid (BA) and oleanolic acid (OA) exhibit considerable pharmacological activities. However, their current production primarily relies on plant extraction and chemical synthesis, methods that are often plagued by low efficiency, complex extraction processes, and environmental concerns. Microbial-based synthesis has emerged as one of the most effective approaches for producing BA and OA. Results This study presented the first identification of filamentous fungal strains efficiently synthesizing both BA and OA. The strain ZBS49 is a newly identified species of Clonostachys isolated from Gleditsia japonica Miq. (showing 99.82% sequence identity), produced 47.7 mg/L of BA. The strain XJ1-1, characterized as Colletotrichum gloeosporioides and isolated from Cannabis sativa L., yielded 65.76 mg/L of OA. After optimizing the culture medium and cultivation conditions, the yields of ZBS49 and XJ1-1 increased to 288.97 and 86.14 mg/L, representing improvements of 506% and 31%, respectively. Furthermore, we discovered that the BA extract of the ZBS49 strain significantly inhibited hepatocellular cancer cells (SMMC-7721 and HepG2) in a dose-dependent manner, with a minimum inhibitory concentration of 70 µM. Genomic analysis of Clonostachys sp. ZBS49 elucidated that the presence of 16 putative genes was related to triterpenoid biosynthesis and 6 distinct terpene biosynthetic gene clusters. Among the 145 CYP450, 5 genes involved in C-28 oxidation were predicted. Conclusions This research underscores the effectiveness of filamentous fungi as a biotechnological platform for the efficient production of BA and its derivatives, highlighting their potential applications in cancer therapy. Furthermore, these findings provide valuable genetic resources and establish a robust technical and theoretical framework for utilizing ZBS49 as a microbial platform for the biosynthesis of triterpenoids. Graphical abstract |
| format | Article |
| id | doaj-art-9685baa48b8541ae99c796b85e4fc460 |
| institution | OA Journals |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-9685baa48b8541ae99c796b85e4fc4602025-08-20T02:05:42ZengBMCMicrobial Cell Factories1475-28592025-06-0124111710.1186/s12934-025-02733-wA new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cellsLuran Geng0Jiale Cui1Changyixin Xiao2Linlin Xu3Fengjiao Yue4Ting Zhang5Mengran Zhan6Zichang Lu7Yuzhe Ren8Chunsheng Wang9Jing Yin10College of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Aulin, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityCollege of Life Science, Northeast Forestry UniversityKey Laboratory of Saline-alkali Vegetation Ecology Restoration, Ministry of Education (Northeast Forestry University)Abstract Background Triterpenoid compounds such as betulinic acid (BA) and oleanolic acid (OA) exhibit considerable pharmacological activities. However, their current production primarily relies on plant extraction and chemical synthesis, methods that are often plagued by low efficiency, complex extraction processes, and environmental concerns. Microbial-based synthesis has emerged as one of the most effective approaches for producing BA and OA. Results This study presented the first identification of filamentous fungal strains efficiently synthesizing both BA and OA. The strain ZBS49 is a newly identified species of Clonostachys isolated from Gleditsia japonica Miq. (showing 99.82% sequence identity), produced 47.7 mg/L of BA. The strain XJ1-1, characterized as Colletotrichum gloeosporioides and isolated from Cannabis sativa L., yielded 65.76 mg/L of OA. After optimizing the culture medium and cultivation conditions, the yields of ZBS49 and XJ1-1 increased to 288.97 and 86.14 mg/L, representing improvements of 506% and 31%, respectively. Furthermore, we discovered that the BA extract of the ZBS49 strain significantly inhibited hepatocellular cancer cells (SMMC-7721 and HepG2) in a dose-dependent manner, with a minimum inhibitory concentration of 70 µM. Genomic analysis of Clonostachys sp. ZBS49 elucidated that the presence of 16 putative genes was related to triterpenoid biosynthesis and 6 distinct terpene biosynthetic gene clusters. Among the 145 CYP450, 5 genes involved in C-28 oxidation were predicted. Conclusions This research underscores the effectiveness of filamentous fungi as a biotechnological platform for the efficient production of BA and its derivatives, highlighting their potential applications in cancer therapy. Furthermore, these findings provide valuable genetic resources and establish a robust technical and theoretical framework for utilizing ZBS49 as a microbial platform for the biosynthesis of triterpenoids. Graphical abstracthttps://doi.org/10.1186/s12934-025-02733-wFilamentous fungiBetulinic acidFermentation systemClonostachys Sp.GenomeTumor suppression |
| spellingShingle | Luran Geng Jiale Cui Changyixin Xiao Linlin Xu Fengjiao Yue Ting Zhang Mengran Zhan Zichang Lu Yuzhe Ren Chunsheng Wang Jing Yin A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells Microbial Cell Factories Filamentous fungi Betulinic acid Fermentation system Clonostachys Sp. Genome Tumor suppression |
| title | A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells |
| title_full | A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells |
| title_fullStr | A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells |
| title_full_unstemmed | A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells |
| title_short | A new Clonostachys Sp. ZBS49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells |
| title_sort | new clonostachys sp zbs49 filamentous fungus with high production of betulinic acid and its inhibitory effect on liver cancer cells |
| topic | Filamentous fungi Betulinic acid Fermentation system Clonostachys Sp. Genome Tumor suppression |
| url | https://doi.org/10.1186/s12934-025-02733-w |
| work_keys_str_mv | AT lurangeng anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT jialecui anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT changyixinxiao anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT linlinxu anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT fengjiaoyue anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT tingzhang anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT mengranzhan anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT zichanglu anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT yuzheren anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT chunshengwang anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT jingyin anewclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT lurangeng newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT jialecui newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT changyixinxiao newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT linlinxu newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT fengjiaoyue newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT tingzhang newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT mengranzhan newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT zichanglu newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT yuzheren newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT chunshengwang newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells AT jingyin newclonostachysspzbs49filamentousfunguswithhighproductionofbetulinicacidanditsinhibitoryeffectonlivercancercells |