Characterization of antibacterial compounds produced by psychrotrophic Alcaligenes faecalis HTP6 isolated from Passu glacier, Pakistan

Paper Details

Research Paper 01/05/2016
Views (252) Download (15)

Characterization of antibacterial compounds produced by psychrotrophic Alcaligenes faecalis HTP6 isolated from Passu glacier, Pakistan

Muhammad Rafiq, Muhammad Hayat, Noor Hassan, Muhammad Ibrar, Abdul Haleem, Maliha Rehman, Faisal Ahmad, Aamer Ali Shah, Fariha Hasan
Int. J. Biosci.8( 5), 122-135, May 2016.
Certificate: IJB 2016 [Generate Certificate]


Low temperature microorganisms can produce secondary metabolites including anticancer, antiviral and antibacterial compounds. The purpose of the study was to evaluate the possibility of using cold adapted bacteria for production of antibiotics. In the present research work, bacteria were isolated from sediment sample, collected from Passu glacier, using R2A medium . These isolates were screened for their resistance towards various antibiotics, antimicrobial activity against P. aeruginosa, E. coli, S. aureus, E. faecalis, C. albicans and A. fumigatus and storage stability of crude extract along with cytotoxicity and haemolytic activity. The isolate HTP6 showed best inhibition using spot on lawn test and was selected for further study. The isolate HTP6 was Gram positive, non-pigmented rod, moderate halophilic, with optimum growth at mesophilic range and was identified as Alcaligenes faecalis HTP6 on the basis of 16S rRNA gene sequence analysis. The strain showed resistance to clindamycin, cefotaxime and sulfamethoxazole/trimethoprim and was able to inhibit P. aeruginosa, E. coli, S. aureus, E. faecalis, C. albicans and A. fumigatus. Maximum growth and inhibitory activity of Alcaligenes faecalis HTP6 was observed against selected ATCC strains [Staphylococcus aureus (ATCC 25923) and Pseudomonas aeruginosa (ATCC 27853)] and various clinical isolates (S. aureus, E. faecalis, Candida albicans and Aspergillus fumigatus) at pH 7 and 30°C, when LB (Luria Bertani) and LB1 (medium supplemented by FeSO4) broth media were used. The crude extract showed good storage and thermal stability at 55°C, and pH stability at 7 along with brine shrimp lethality up to 30%, however, there was no DNA binding and haemolytic activity observed. We can conclude from the study that Alcaligenes faecalis HTP6, isolated from Passu glacier, can be a good candidate for the production of wide spectrum potent thermostable antibiotic with less cytotoxicity.


Al-Judaibi A. 2011.  Effect  of  some  fermentation parameters on ethanol production from beet molasses by Saccharomyces Cerevisiae CAIM13. American Journal of Agriculture and Biological Sciences 6, 301-306.

Al-zereini W, Schuhmann I, Laatsch H, Helmke E, Anke H. 2007.  New  aromatic  nitro compounds from Salegentibacter sp. T436, an Arctic sea ice bacterium: Taxonomy, fermentation, isolation and biological activities. Journal of Antibiotics 60, 301-308.

Anesio AM, Laybourn-Parry J. 2012.  Glaciers and ice sheets as a biome. Trends in Ecology and Evolution 27, 219-225.

Asencio G, Lavin P, Alegría K. 2014. Antibacterial activity of the Antarctic bacterium Janthinobacterium sp: SMN 33.6 against multi-resistant Gram-negative bacteria. Electronic Journal of Biotechnology 17, 1-1.

Bruntner C, Binder T, Pathom-aree W. 2005. Frigocyclinone, a novel angucyclinone antibiotic produced by a Streptomyces griseus strain from Antarctica. Journal of Antibiotics 58, 346-349.

De-Souza MJ, Nair SL, Bharathi PA, Chandramohan D. 2006. Metal and antibiotic-resistance in psychrotrophic bacterial from Antarctic marine waters. Ecotoxicology 15, 379-384.

Feller G, Gerday C. 2003. Psychrophilic enzymes: hot  topics  in  cold  adaptation.  Nature  Reviews  in Microbiology 1, 200-208.

Garrity GM, Bell JA, Lilburn TG. 2004. Taxonomic outline of the prokaryotes, In: Bergey’s manual of systematic bacteriology 2nd Edition. Springer Verlag, New York.

Giudice AL, Casella P, Bruni V, Michaud L. 2013. Response of bacterial isolates from Antarctic shallow sediments towards heavy metals, antibiotics and polychlorinated biphenyls. Ecotoxicology 22, 240-250.

Hemala L, Zhang D, Margesin R. 2014. Cold-active antibacterial and antifungal activities and antibiotic resistance of bacteria isolated from an alpine hydrocarbon-contaminated industrial site. Research in Microbiology 165, 447-456.

Honda N, Hirai M, Ano T, Shoda M. 1998. Antifungal effect of a heterotrophic nitrifier Alcaligenes faecalis. Biotechnology Letters 20, 703-705.

Jayanth N. 2001. Air-conditioning servicing system and method. Google Patents.

Kapley A, Sagarkar S, Tanksale H. 2013. Genome sequence of Alcaligenes sp. strain HPC1271. Genome Announcement 10, 00235-12.

Kay S, Cheeptham N. 2013. Screening for antimicrobial activities of cave actinomycetes against honeybee pathogen. Chiang Mai Journal of Science 40, 26–33.

Li ZY, Peng C, Shen Y, Miao X, Zang H, Lin H. 2008. L,L Diketopiperazines from Alcaligenes faecalis A72 associated with South China Sea sponge Stelletta tenuis. Biochemical Systematics and Ecology 36, 230-234.

Li M, Cha DJ, Lai Y, Villaruz AE, Sturdevant DE, Otto M. 2007. The antimicrobial peptide‐sensing system aps of Staphylococcus aureus. Molecular Microbiology 66, 1136-1147.

Maridass M. 2008. Evaluation of Brine Shrimp Lethality of Cinnamomum species. Ethnobotany Leaflets 12, 772-775.

Mangano S, Luigi M, Consolazione C, Angelina LG. 2014. Metal and antibiotic resistance in psychrotrophic bacteria associated with the Antarctic sponge Hemigellius pilosus (Kirkpatrick, 1907). Polar Biology 37, 227-235.

Margesin R, Miteva V. 2011. Diversity and ecology of psychrophilic microorganisms. Research in Microbiology 162, 346-361. doi:10.1016/j.resmic.2010.12.004

Martinez C, Avis TJ, Simard JN. 2006. The role of antibiosis in the antagonism of different bacteria towards Helminthosporium solani, the causal agent of potato silver scurf. Phytoprotection 87, 69-76.

Morgan-kiss RM, Priscu JC, Pocock T, Gudynaite-savitch l, Huner NP. 2006. Adaptation and acclimation of photosynthetic microorganisms to permanently cold environments. Microbiology and Molecular Biology Reviews 70, 222-252.

Morita Y. 1975. Psychrophilic bacteria. Bacteriology Reviews 39, 144-167.

O’Brien AR, Sharp J. Nicholas S, Roller. 2004. Antarctic bacteria inhibit growth of food-borne microorganisms at low temperatures. FEMS Microbiology Ecology 48, 157-167.

Omura S, Ikeda H, Malpartida F, Kieser HM, Hopwood  DA.  1986.  Production  of  new  hybrid antibiotics, mederrhodins A and B, by a genetically engineered strain. Antimicrobial Agents and Chemotherapy 29, 13-19.

Ravot G, Masson JM, Lefèvre F. 2006. 34 applications of extremophiles: the industrial screening of extremophiles for valuable biomolecules. Methods in Microbiology 35, 785-813.

Ripa FA, Nikkon F, Zaman S, Khondka P. 2009. Optimal conditions for antimicrobial metabolites production from a new Streptomyces sp. RUPA-08PR isolated from Bangladeshi soil. Mycobiology 37, 211–214.

Rodrigues DF, Tiedje JM. 2008. Coping with our cold planet. Applied and Environmental Microbiology 74, 1677-1686.

Russell NJ, Harrisson P, Johnston IA. 1990. Cold adaptation of microorganisms and discussion. Philosophical  Transactions  of  the  Royal Society of London B: Biological Sciences 326, 595-611.

Sajjad W, Rafiq M, Zada S. 2015. Phylogenetic analysis of newly isolated protease producing salt tolerant psychrophilic bacteria from Tirich Mir glacier, Pakistan. International Journal of Biosciences 7, 159-169.

Sanchez LA, Gómez FF, Delgado OD. 2009. Cold-adapted microorganisms as a source of new antimicrobials. Extremophiles 13, 111-120.

Sanchez S, Adan C, Angela F et al. 2010. Carbon source regulation of antibiotic production. The Journal of Antibiotics 63, 442-459.

Shekh, RM, Singh P, Singh SM. 2011. Antifungal activity of Arctic and Antarctic bacteria isolates. Polar Biology 34, 139-143.

Talbot GH, Bradley J, Edwards JE. 2006. Bad bugs need drugs: an update on the development pipeline from the antimicrobial availability task force of the Infectious Diseases Society of America. Clinical and Infectious Disease 42, 657-668.

Tamura K, Nei M. 1993. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution 10, 512-526.

Tamura K, Stecher G, Peterson D. 2013. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution 30, 2725-2729.

Tehei M, Madern D, Franzetti B, Zaccai G. 2005. Neutron scattering reveals the dynamic basis of protein adaptation to extreme temperature. Journal of Biological Chemistry 280, 40974-40979.

Usha KM, Sudhakar P, Sreenivasulu K, Vijayalakshmi M. 2011. Optimization of culturing conditions for improved production of bioactive metabolites by Pseudonocardia sp. VUK-10 Mycobiology 39, 174-181.

Walsh C. 2003. Where will new antibiotics come from? Nature Reviews in Microbiology 1, 65-70.

Yegneswaran PK, Gray MR, Westlake DWS. 1988. Effects of reduced oxygen on growth and antibiotic production in Streptomyces clavuligerus. Biotechnology Letters 10, 479–84.

Zahir I, Houari A, Bahafid W. 2013. A novel Alcaligenes faecalis antibacterial-producing strain isolated from a Moroccan tannery waste. African Journal of Microbiology Research 7, 5314-5323.