Screening, Identification and Antitumor Activity of <em>Myxococcus stipitatus</em> STXZ77

doi:10.13523/j.cb.20161102

China Biotechnology

2016, Vol. 36

Issue (11): 7-15 DOI: 10.13523/j.cb.20161102

Screening, Identification and Antitumor Activity of Myxococcus stipitatus STXZ77

LEI Liang huan, HUANG Tong long, WEI Hui, LIAO Ji yan, WU Yu jing, ZHOU Cai, XIA Li qiu, ZHANG You ming

State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha 410081, China

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Abstract

Objective:Screening and identifying the myxobacteria from soil samples to enrich myxobacteria resources, then preliminarily study on its antitumor activity, to lay the foundation for the exploration of anti-tumor drugs. Methods:Isolating myxobacteria strains from soil samples through inactivated Escherichia coli inducing method, identifying the strain based on morphological observation, physiologically and biochemically characterizing, and the homologous analysis of 16S rRNA sequences of nucleotides. XAD-16 macroporous adsorption resin was added to fermented supernatant to harvest crude extracting from fermented product, and evaluated its antitumor activity in vitro through CCK-8 assay. An antitumor active component was isolated from the identified strain STXZ77 by reversed phase high-performance liquid chromatography (RP-HPLC) technique, and its molecular weight was analyzed by LC-MS/MS. Results:The strain STXZ77, which was isolated and identified, was named Myxococcus stipitatus STXZ77. The XAD-16 resin crude extract from the fermented products expressed antitumor activity against many kinds of tumor cells, such as mouse melanoma cells B16, mouse mammary cells 4T1, human hepatoma cells SMMC-7721, human cervical carcinoma cells HeLa, human colon cells SW480, etc. The IC₅₀ values are 5.34μg/ml, 13.50μg/ml, 11.93μg/ml, 28.70μg/ml and 48.09μg/ml, respectively. While it showed minor toxicity to HUVEC cell line, the IC₅₀ value of which is 17.09μg/ml, indicating less toxicity compared with B16, 4T1 and SMMC-7721 cell lines. An antitumor active component, AP-C, was successfully isolated using RP-HPLC technique, the mass-to-charge ratio of which was determined to be 422.99m/z through mass spectrometry analysis. Conclusion:The myxobacterium strain Myxococcus stipitatus STXZ77 was isolated from soil samples, from which an antitumor active compound AP-C was obtained, and it has a significant value for developing antitumor drugs.

Key words： Antitumor Screening STXZ77 Myxobacterium Purification

Received: 03 May 2016 Published: 25 November 2016

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LEI Liang huan, HUANG Tong long, WEI Hui, LIAO Ji yan, WU Yu jing, ZHOU Cai, XIA Li qiu, ZHANG You ming. Screening, Identification and Antitumor Activity of Myxococcus stipitatus STXZ77. China Biotechnology, 2016, 36(11): 7-15.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20161102 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I11/7

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