Partially Purified Pigment Extract from Streptomyces A 16-1 Induces Apoptosis of Human Carcinoma of Nasopharynx Cell (KB cells) via the Mitochondrial and Caspase-3 Pathway

Chantarawan SAENGKHAE, Rattanaporn SRIVIBOOL, Rawiwan WATANADILOK, Keiichi ENOMOTO


The genus Streptomycetes are very important bacteria which produce secondary metabolites used in clinical cancer therapy and other pharmacological activities. Streptomyces A 16-1 was isolated from coastal soil in Chonburi, Thailand. The objective of this study is to examine the apoptotic effects and mechanisms of the partially purified pigment extract from Streptomyces A 16-1 against human carcinoma of nasopharynx cell (KB cells). The methanol and ethyl acetate extract from the fermentation of ISP2 broth was fractionated by using column chromatography with stationery-phase silica-gel and mobile phase chloroform: methanol mixtures with increasing polarity. Among 9 fractions (Fr), the fractions which contained red pigment were selected and the calculated IC50 values for Fr 5, Fr 6 and Fr 7 were 0.04 ± 0.005, 0.20 ± 0.02, and 0.55 ± 0.05 μg/ml, respectively. The molecular mechanisms of cell death were associated with DNA fragmentation, accumulation of cells in sub-G1 phase, mitochondrial membrane depolarization, and activation of caspase-3, which was significantly diminished by the caspase-3 inhibitor. These first findings demonstrate that the pro-apoptotic effects of these fractions were mediated through the mitochondrial and caspase-3 pathway.


Apoptosis, Streptomyces A 16-1, DNA fragmentation, mitochondria, caspase-3

Full Text:



S Ramesh, N Mathivanan. Screening of marine actinomycetes isolated from the Bay of Bengal, India for antimicrobial activity and industrial enzymes. World J. Microbiol. Biotechnol. 2009; 25, 2103-11.

C Balachandran, Y Arun, V Duraipandiyan, S Ignacimuthu, K Balakrishna and NA Al-Dhabi. Antimicrobial and cytotoxicity properties of 2,3-dihydroxy-9,10-anthraquinone isolated from Streptomyces galbus (ERINLG-127). Appl. Biochem. Biotechnol. 2014; 172, 3513-28.

AF Braña, HP Fiedler, H Nava, V González, A Sarmiento-Vizcaíno, A Molina, JL Acuña, LA García and G Blanco. Two Streptomyces species producing antibiotic, antitumor, and anti-inflammatory compounds are widespread among intertidal macro algae and deep-sea coral reef invertebrates from the central Cantabrian Sea. Microb. Ecol. 2015; 69, 512-24.

S Sudha and MM Selvam. Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4. Asian Pac. J. Trop. Biomed. 2012; 2, 770-3.

ST Williams, ME Sharpe and JG Holt. Bergey’s Manual of Systematic Bacteriology. Vol IV, Williams & Wilkins Press, Baltimore, 1989, p. 2648.

C Olano, C Méndez and JA Salas. Antitumor compounds from marine actinomycetes. Mar. Drugs 2009; 7, 210-48.

H Zhang, H Wang, H Cui, Z Li, Z Xie, Y Pu, F Li, S Qin. A new anthracene derivative from marine Streptomyces sp. W007 exhibiting highly and selectively cytotoxic activities. Mar. Drugs. 2011; 9, 1502-1509.

Z Xie, B Liu, H Wang, S Yang, H Zhang, Y Wang, N Ji, S Qin and H Laatsch. Kiamycin, a unique cytotoxic angucyclinone derivative from a marine Streptomyces sp. Mar. Drugs 2012; 10, 551-8.

C Park, HJ Shin, GY Kim, TK Kwon, TJ Nam, SK Kim, J Cheong, IW Choi and YH Choi. Induction of apoptosis by streptochlorin isolated from Streptomyces sp. in human leukemic U937 cells. Toxicol. In Vitro 2008; 22, 1573-81.

SD Yeok, HJ Shin, GY Kim, JH Cheong, IW Choi, SK Kim, SK Moon, HS Kang and YH Choi. Streptochlorin isolated from Streptomyces sp. induces apoptosis in human hepatocarcinoma cells through a reactive oxygen species-mediated mitochondrial pathway. J. Microbiol. Biotechnol. 2008; 18, 1862-7.

V Karuppiah, C Aarthi, K Sivakumar and L Kannan. Statistical optimization and anticancer activity of a red pigment isolated from Streptomyces sp. PM4. Asian Pac. J. Trop. Biomed. 2013; 3, 650-6.

DN Raj, D Dhanasekaran, N Thajuddin and A Panneerselvam. Production of prodigiosin from Serratia marcescens and its cytotoxicity activity. J. Pharm. Res. 2009; 2, 590-3.

S Elmore. Apoptosis: A review of programmed cell death. Toxicol. Pathol. 2007; 35, 495-516.

JD Ly, DR Grubb and A Lawen. The mitochondrial membrane potential (Δψm) in apoptosis; An update. Apoptosis 2003; 8, 115-28.

EB Shirling and D Gottlieb. Methods for characterization of Streptomycetes species. Int. J. Syst. Bacteriol. 1966; 13, 313-23.

M Anibou, A Chait, A Zyad, M Taourirt, Y Ouhdouch and A Benherref. Actinomycetes from Moroccan habitats: Isolation and screening for cytotoxic activities. World J. Microbiol. Biotechnol. 2008; 24, 2019-25.

C Saengkhae, Y Premsuriya, R Sriviboon and J Praiboon. Sensitization of human carcinoma of nasopharynx cells to doxorubicin and induction of apoptosis by Sargassum baccularia lipophilic fraction. Walailak J. Sci. & Tech. 2015; 12, 515-25.

M Suffness and JM Pezzuto. Assays Related to Cancer Drug Discovery. In: K Hostettmann (ed.). Methods in Plant Biochemistry: Assays for Bioactivity. Vol VI. Academic Press, London, 1990, p. 71-133.

RB Weiss. The anthracyclines: Will we ever find a better doxorubicin? Seminars Oncol. 1992; 19, 670-86.

TL Ecuyer, S Sanjeev, R Thomas, R Novak, L Das, W Campbell and V Heide. DNA damage is an early event in doxorubicin-induced cardiac myocyte death. Am. J. Physiol. Heart Circ. Physiol. 2006; 29, H1273-H1280.

C Balachandran, B Sangeetha, V Duraipandiyan, MK Raj, S Ignacimuthu, NA Al-Dhabi, K Balakrishna, K Parthasarathy, NM Arulmozhi and MV Arasu. A flavonoid isolated from Streptomyces sp. (ERINLG-4) induces apoptosis in human lung cancer A549 cells through p53 and cytochrome c release caspase dependent pathway. Chem. Biol. Interact. 2014; 224, 24-35.

CH Lee, H Lim, S Moon, C Shin, S Kim, BJ Kim and Y Lim. Novel anticancer agent, benzyldihydroxyoctenone, isolated from Streptomyces sp. causes G1 cell cycle arrest and induces apoptosis of HeLa cells. Cancer Sci. 2007; 98, 795-802.

H Liu, L Xiao, J Wei, JC Schmitz, M Liu, C Wang, L Cheng, N Wu, L Chen, Y Zhang and X Lin. Identification of Streptomyces sp. nov. WH26 producing cytotoxic Streptomycetes compounds isolated from marine solar saltern in China. World J. Microbiol. Biotechnol. 2013; 29, 1271-8.

LE Gerschenson and R Rotello. Apoptosis: A different type of cell death. Fed. Am. Soc. Exp. Biol. J. 1992; 6, 2450-5.

AI Scovassi and A Torriglia. Activation of DNA-degrading enzymes during apoptosis. Eur. J. Histochem. 2003; 47, 185-94.

S Lee, M Baek, HY Kim, JH Ha and DI Jeoung. Mechanism of doxorubicin-induced cell death and expression profile analysis. Biotechnol. Lett. 2002; 24, 1147-51.

JH Lee, C Zhang, JY Ko, JS Lee and YJ Jeon. Evaluation on anticancer effect against HL-60 cells and toxicity in vitro and in vivo of the phenethyl acetate isolated from a marine bacterium Streptomyces griseus. Fish Aquat. Sci. 2015; 18, 35-44.

V Rambabu, S Suba, P Manikanda and S Vijayakumar. Cytotoxic and apoptotic nature of migrastatin, a secondary metabolite from evaluated on HepG2 cell line. Int. J. Pharm. Sci. 2014; 6, 333-8.

V Rambabu, S Suba and S Vijayakumar. Antimicrobial and antiproliferative prospective of kosinostatin a secondary metabolite isolated from Streptomyces sp. J. Pharm. Anal. 2015; 5, 378-82.


  • There are currently no refbacks.

Online ISSN: 2228-835X

Last updated: 20 June 2019