|
|
Diversity, Novelty and Anti-tumor Activity of Actinomyces from Coastal Mangrove in Leizhou, Zhanjiang |
LI Jun-Jie1, LIN Man-Jia4, CHEN Hai-Ming1,2,3, LIU Yue-Li1, LIANG Xiao-Yan1, HUANG Yong-Mei1,2,3,*, LUO Hui1,2,3,* |
1 Marine Biomedical Research Institute, the Key Lab of Zhanjiang for R&D Marine Microbial Resources in the Beibu Gulf Rim, Guangdong Medical University, Zhanjiang 524023, China; 2 Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang 524023, China; 3 The Marine Biomedical Research Institute of Guangdong Zhanjiang, Zhanjiang 524023, China; 4 First Clinical Medical College, Guangdong Medical University, Zhanjiang, 524023, China |
|
|
Abstract Most of mangroves grow in intertidal zones with special and changeable environments, which are conducive for the growth of rare Actinomyces and production of active compounds. In order to explore the diversity, novelty and medicinal potential of Actinomyces resources from coastal mangrove in Leizhou, Zhanjiang, China. Pour plate method and 10 different media were used in this study to isolate Actinomyces from mangrove rhizosphere soil samples. After purification and culture, PCR amplification and sequencing and phylogenetic analysis of 16S rRNA of Actinomyces strains were performed. At the same time, according to the comparison results of 16S rRNA and the colony morphological characteristics of strains, portion of Actinomyces strains were selected for small-scale fermentation, all broths without mycelia were extracted by ethyl acetate and dried by rotary evaporator before the preparation of fermentation crude extract methanol solution. Anti-tumor activity of fermentation crude extracts against human lung cancer cell line A549 and human liver cancer cell line HepG2 were analyzed by Cell Counting Kit-8 (CCK-8). According to the result, a total of 154 Actinomycetes were isolated from 8 rhizosphere soil samples collected from 3 different sampling sites in coastal areas of Zhanjiang Mangrove National Nature Reserve, Guangdong Province, distributing in 11 orders, 15 families and 19 genera, with the dominant genera were Micromonspora (22.7%, 35 strains), followed by Microbacterium (20.8%, 32 strains) and Streptomyces (10.4%, 16 strains). And there were 7 potential new species distributing in 4 genera with 16S rRNA sequence similarity less than 98.65%. In the anti-tumor activity screening of fermentation crude extracts, 53 strains of Actinomyces showed anti-tumor activity against at least one tumor cell line, accounting for 79.1% of 67 strains. The above results suggested that, the diversity of Actinomyces and potential medicinal resources were rich in coastal mangrove areas of Leizhou. This study bases a sufficient strain foundation for the further improvement of the cognition of Actinomyces distribution in Leizhou coastal areas as well as the subsequent research on other biological activities such as antibacterial activities and related mechanisms.
|
Received: 11 August 2022
|
|
Corresponding Authors:
*huangym@gdmu.edu.cn; luohui@gdmu.edu.cn
|
|
|
|
[1] 但新球, 廖宝文, 吴照柏, 等. 2016. 中国红树林湿地资源、保护现状和主要威胁[J]. 生态环境学报, 25(07): 1237-1243. (Dan X Q, Liao B W, Wu Z B, et al.2016. Resources, conservation status and main threats of mangrove wetlands in China[J]. Ecology and Environment, 25(07): 1237-1243.) [2] 方雅. 2019. 三株具有抗肿瘤活性放线菌次级代谢产物的研究[D]. 硕士学位论文, 厦门大学, 导师: 邓贤明; 徐庆妍, pp. 152. (Fang Y.2019. Study on secondary metabolites of three Actinomycetes with anti-tumor activity[D]. Thesis for M.S., Xiamen University, Supervisor: Deng X M; XU Q Y, pp. 152.) [3] 李蜜, 易湘茜, 杨彩妮, 等. 2020. 海南西海岸红树林伴生植物内生放线菌多样性及其延缓衰老活性初筛[J]. 广西植物, 40(03): 293-300. (Li M, Yi X Q, Yang C N, et al.2020. Diversity and anti-aging activity of endophytic Actinobacteria from associated mangrove plants collected from west coast of Hainan[J]. Guihaia, 40(03): 293-300.) [4] 李逾. 2012. 三种红树植物共附生放线菌的分离及其抗病毒活性菌株的筛选[D]. 硕士学位论文, 海南大学, 导师: 鲍时翔, pp. 69. (Li Y.2012. Isolation and screening anti-viral Actinomycetes from three mangrove[D]. Thesis for M.S., Hainan University, Supervisor: Bao S X, pp. 69.) [5] 唐秋霞, 王友绍. 2021. 雷州半岛红树林群落特征及其分布格局[J]. 生态科学, 40(05): 23-32. (Tang Q X, Wang Y S.2021. Characteristics and distribution pattern of mangrove community in the Leizhou peninsula[J]. Ecological Science, 40(05): 23-32.) [6] 向晨晨, 周珊珊, 柴树茂, 等. 2021. 红树林链霉菌ZFSM1-146中抗菌活性物质的发现[J]. 微生物学通报, 48(07): 2329-2340. (Xiang C C, Zhou S S, Chai S M, et al.2021. Discovery of antibacterial compounds from a mangrove Streptomyces strain ZFSM1-146[J]. Microbiology China, 48(07): 2329-2340.) [7] 许敏, 李静, 戴素娟, 等. 2016. 广东湛江红树林植物内生放线菌资源勘探及生物活性研究[J]. 中国抗生素杂志, 41(01): 26-34. (Xu M, Li J, Dai S J, et al.2016. Study on diversity and bioactivity of Actinobacteria isolated from mangrove plants collected from Zhanjiang in Guangdong Province[J]. Chinese Journal of Antibiotics, 41(01): 26-34.) [8] 尹周一, 王梦圆, 游伟程, 等. 2022. 2022美国癌症统计报告解读及中美癌症流行情况对比[J]. 肿瘤综合治疗电子杂志, 8(02): 54-63. (Yin Z Y, Wang M Y, You W C, et al.2022. Interpretation on the report of American cancer statistics, 2022 and comparison of cancer prevalence in China and America[J]. Journal of Multidisciplinary Cancer Management (Electronic Version), 8(02): 54-63.) [9] 赵梦冉, 王璐, 叶伟霞, 等. 2022. 湛江高桥红树林沉积物放线菌多样性及其中一株链霉菌产生的germicidins类化合物的鉴定[J]. 微生物学报, 62(05): 1740-1753. (Zhao M R, Wang L, Ye W X, et al.2022. Diversity of Actinobacteria from mangrove sediment in Gaoqiao, Zhanjiang and identification of germicidins from one strain of Streptomyces[J]. Acta Microbiologica Sinica, 62(05): 1740-1753.) [10] 周双清, 黄小龙, 黄东益, 等. 2010. Chelex-100快速提取放线菌DNA作为PCR扩增模板[J]. 生物技术通报, 211(02): 123-125. (Zhou S Q, Huang X L, Huang D Y, et al.2010. A rapid method for extracting DNA from Actinomycetes by Chelex-100[J]. Biotechnology Bulletin, 211(02): 123-125.) [11] Barbier P, Guise S, Huitorel P, et al.2001. Caulerpenyne from Caulerpa taxifolia has an antiproliferative activity on tumor cell line SK-N-SH and modifies the microtubule network[J]. Life Sciences, 70(4): 415-429. [12] Chao C H, Wen Z H, Wu Y C, et al.2008. Cytotoxic and anti-inflammatory cembranoids from the soft coral Lobophytum crassum[J]. Journal of Natural Products, 71(11): 1819-1824. [13] Chen L, Chai W, Wang W, et al.2017. Cytotoxic bagremycins from mangrove-derived Streptomyces sp. Q22[J]. Journal of Natural Products, 80(5): 1450-1456. [14] Cho J Y, Kwon H C, Williams P G, et al.2006. Actinofuranones A and B, polyketides from a marine-derived bacterium related to the genus Streptomyces (Actinomycetales)[J]. Journal of Natural Products, 69(3): 425-428. [15] Cho J Y, Williams P G, Kwon H C, et al.2007. Lucentamycins A-D, cytotoxic peptides from the marine-derived Actinomycete Nocardiopsis lucentensis[J]. Journal of Natural Products, 70(8): 1321-1328. [16] Cui C B, Liu H B, Gu J Y, et al.2007. Echinosporins as new cell cycle inhibitors and apoptosis inducers from marine-derived Streptomyces albogriseolus[J]. Fitoterapia, 78(3): 238-240. [17] Du L, Feng T, Zhao B, et al.2010. Alkaloids from a deep ocean sediment-derived fungus Penicillium sp. and their antitumor activities[J]. The Journal of Antibiotics, 63(4): 165-170. [18] Feling R H, Buchanan G O, Mincer T J, et al.2003. Salinosporamide A: A highly cytotoxic proteasome inhibitor from a novel microbial source, a marine bacterium of the new genus Salinospora[J]. Angewandte Chemie, 42(3): 355-357. [19] Hu C, Zhou S W, Chen F, et al.2017. Neoantimycins A and B, two unusual benzamido nine-membered dilactones from marine-derived Streptomyces antibioticus H12-15[J]. Molecules, 22(4): 557. [20] Kimura M.1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences[J]. Journal of Molecular Evolution, 16(2): 111-120. [21] Kumar S, Stecher G, Li M, et al.2018. MEGA X: Molecular evolutionary genetics analysis across computing platforms[J]. Molecular Biology and Evolution, 35(6): 1547-1549. [22] Lu Q P, Huang Y M, Liu S W, et al.2021. Metabolomics tools assisting classic screening methods in discovering new antibiotics from mangrove actinomycetia in Leizhou peninsula[J]. Marine Drugs, 19(12): 688. [23] Matsumoto A, Nakai K, Morisaki K, et al.2010. Demequina salsinemoris sp. nov., isolated on agar media supplemented with ascorbic acid or rutin[J]. International Journal of Systematic and Evolutionary Microbiology, 60(5): 1206-1209. [24] Mawlankar R R, Mual P, Sonalkar V V, et al.2015. Microbacterium enclense sp. nov., isolated from sediment sample[J]. International Journal of Systematic and Evolutionary Microbiology, 65(7): 2064-2070. [25] Miller T W, Chaiet L, Cole D J, et al.1979. Avermectins, new family of potent anthelmintic agents: Isolation and chromatographic properties[J]. Antimicrobial Agents and Chemotherapy, 15(3): 368-371. [26] Mitchell S S, Nicholson B, Teisan S, et al.2004. Aureoverticillactam, a novel 22-atom macrocyclic lactam from the marine Actinomycete Streptomyces aureoverticillatus[J]. Journal of Natural Products, 67(8): 1400-1402. [27] Nouioui I, Carro L, Sangal V, et al.2018. Formal description of Mycobacterium neglectum sp. nov. and Mycobacterium palauense sp. nov., rapidly growing Actinobacteria[J]. Antonie Van Leeuwenhoek, 111(7): 1209-1223. [28] Sagar S, Esau L, Holtermann K, et al.2013. Induction of apoptosis in cancer cell lines by the Red Sea brine pool bacterial extracts[J]. BMC Complementary and Alternative Medicine, 13: 344. [29] Saitou N, Nei M.1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 4(4): 406-425. [30] Sawadogo W R, Boly R, Cerella C, et al.2015. A survey of marine natural compounds and their derivatives with anti-cancer activity reported in 2012[J]. Molecules, 20(4): 7097-7142. [31] Sun J, Shao J, Sun C, et al.2018. Borrelidins F-I, cytotoxic and cell migration inhibiting agents from mangrove-derived Streptomyces rochei SCSIO ZJ89[J]. Bioorganic & Medicinal Chemistry, 26(8): 1488-1494. [32] Tan C, Etcubanas E, Wollner N, et al.1973. Adriamycin-an antitumor antibiotic in the treatment of neoplastic diseases[J]. Cancer, 32(1): 9-17. [33] Thawai C, Tanasupawat S, Suwanborirux K, et al.2011. Agromyces tropicus sp. nov., isolated from soil[J]. International Journal of Systematic and Evolutionary Microbiology, 61(3): 605-609. [34] Thompson J D, Higgins D G, Gibson T J.1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice[J]. Nucleic Acids Research, 22(22): 4673-4680. [35] Xu D B, Ye W W, Han Y, et al.2014. Natural products from mangrove actinomycetes[J]. Marine Drugs, 12(5): 2590-2613. [36] Xu J.2011. Biomolecules produced by mangrove-associated microbes[J]. Current Medicinal Chemistry, 18(34): 5224-5266. [37] Young C W.1969. Actinomycin and antitumor antibiotics[J]. American Journal of Clinical Pathology, 52(2): 130-137. [38] Zovko A, Novak M, Hååg P, et al.2016. Compounds from the marine sponge Cribrochalina vasculum offer a way to target IGF-1R mediated signaling in tumor cells[J]. Oncotarget, 7(31): 50258-50276. |
|
|
|