Nutritional Requirements of Bacillus thuringiensis During Different Phases of Growth, Sporulation and Germination Evaluated by Plackett-Burman Method

Document Type: Research Note


School of Chemical Engineering, College of Engineering, University of Tehran, P.O. Box 11155-4563 Tehran, I.R. IRAN


The effects of different sources of carbon, nitrogen and some of trace elements on cultivation of Bacillus thuringiensis H14 were studied. Glucose, glycerol, sodium acetate, (NH4)2SO4, corn steep liquor, yeast extract, hydrolyzed casein, Ca2+, Mg2+ and Mn2+ were the main parameters evaluated by Plackett-Burman statistical method. All elements have insignificant effect on germination while spore production was mainly affected by corn steep liquor. Growth was affected by corn steep liquor, MnSO4 and sodium acetate. The following medium has been proposed based on the responses to growth and sporulation of B. thuringiensis H14: glucose, 5 g/L; glycerol, 1 g/L; sodium acetate,0 g/L; (NH4)2SO4, 1 g/L; corn steep liquor, 2 mL/L; yeast extract, 1 g/L; hydrolyzed casein, 1 g/L; CaCl2, 20 mg/L; MgSO4, 50 mg/L; MnSO4 100 mg/L; FeSO4, 0 mg/L.This medium resulted in a maximum spore count.


Main Subjects

1] Andrews G.E., Faust R.M., Wakbiko H., Raymond K.C., The Biotechnology of Bacillus thuringiensis, CRC Crit. Rev. Biotechnol, 6, p. 163 (1987).

[2] Najafloo A., Sarrafzadeh M.H., Gerami, A., Statistical Analysis and Modeling of Sporulation Phase During Fermentation of B. thuringiensis to Study the Effect of Oxygen and Culture Time on its Bioinsecticide Activity., J. Petrol. Chem. Eng., 42(8), p. 1025 (2009).

[3] de Maagd R., Bravo A., Crickmore N., How Bacillus thuringiensis has Evolved Specific Toxins to Colonize the Insect World, Trends in Genetics, 17, p. 193 (2001).

[4] Avignon Rossa C.A., Yantorno M.O., Arcas J.A., Ertola R.J., Organic and Inorganic Nitrogen Source Ratio Effects on Bacillus thuringiensis Var. israelensis Delta-Endotoxin Production, World J. Microbiol. Rev, 6, p. 27 (1990).

[5] Faloci M.M., Yantorno O.M., Marino H.A., Arcas J.A., Ertola R.J., Effect of the Media Composition on the Growth Parameters and Biological Properties of B. thuringiensis Delta-Endotoxin, World. J. Microbiol. Biotech, 6, p. 32 (1990).

[6] Goldberg I., Sneh B., Battat E., Klein D., Optimisation of a Medium for a High Yield Production of Spore-Crystal Preparation of B. thuringiensis Effective Against the Egyptian Cotton Leaf Warm, Biotech. Lett, 2, p. 419 (1980).

[7] Salama H.S., Foda M.S., Dulmage H.T., El-Sharaby A., Novel Fermentation Media for Production of d-Endotoxins from Bacillus thuringiensis, J. Invertebr. Pathol, 41, p. 8 (1983).

[8] Naveena B.J., Altaf M., Bhadriah K., Reddy G., Selection of Medium Components by Plackett-Burman Design for Production of Lactic Acid by L. amylophilus GV6 in SSF Using Wheat Bran, Biores. Tech., 96, p. 485 (2005).

[9] Chauhan K., Trivedi U.B., Patel K.C., Statistical Screening of Medium Components by Plackett-Burman Design for Lactic Acid Production by Lactobacillus sp. KCP01 Using Date Juice, Bioresource Technology, 98, p. 98 (2007).

[10] Robert A. Smith., Effect of Strain and Medium Variation on Mosquito Toxin Production by Bacillus thuringiensis var. israelensis, Can. J. Microbiol, 28(9), p. 1089 (1982).

[11] Sasaki K, Jiaviriyaboonya S, Rogers, P.L., Enhancement of Sporulation and Crystal Production in Culture of Bacillus Sphaericus 2362, Biotechnol Lett, 20, p. 165 (1998).

[12] Yerra K.R., Kuen-Juh T., Wen-Shi W.., Yew-Min T., Medium Optimization of Carbon and Nitrogen Sources for the Production of Spores from B. amyloliquefaciens B128 Using RSM, Process Biochemistry, 42(4), p. 535 (2007).

[13] Vimala Devi P.S., Ravinder T., Jaidev C., Cost-Effective Production of Bacillus thuringiensis by Solid-State Fermentation, Journal of Invertebrate Pathology, 88(2), p. 163 (2005).

[14] Abdel-Hameed A., Carlberg G., El-Tayeb O.M., Studies on B. thuringiensis H-14 Strains Isolated in Egypt-IV. Characterization of Fermentation Conditions for δ-Endotoxin, World J. Microb. Biotech., 7(2), p. 231 (1991).

[15] Liu Y-B., Tabashnik, B.E., Johnson M.W., Larval Age Affects Resistance to Bacillus thuringiensis in Diamondback Moth (Lepidoptera: Plutellidae), J. Econ. Entomol., 88, p. 788 (1995).

[16] Prabakaran G., Balaraman K., Hoti S.L., Manonmani A.M., A Cost-Effective Medium for the Large-Scale Production of B. sphaericus H5a5b for Mosquito Control, Biological Control, 41(3), p. 379 (2007).

[17] Pearson D., Ward OP., Effect of Culture Conditions on Growth and Sporulation of B. thuringiensis Subsp. israelensis and Development of Media for Production of the Protein Crystal Endotoxin, Biotech Lett, 10, p. 451 (1988).

[18] Foda M.S., Salama H.S., Selim M., Factors Affecting Growth Physiology of B. thuringiensis, Applied Microbiology and Biotechnology, 22, p. 50 (1985).

[19] Magda A. El-Bendary, Dr., Bacillus thuringiensis and Bacillus sphaericus Pesticides Production, Journal of Basic Microbiology, 46(2), p. 158 (2006).

[20] Tasharrofi N., Adrangi S., Fazeli M., Rastegar H., Khoshayand M.R., Faramarzi M.A., Optimization of Chitinase Production by Bacillus Pumilus Using Plackett-Burman Design and Response Surface Methodology, Iranian Journal of Pharmaceutical Research, 10 (4), p. 759 (2011)

[21] Hoseyni S.M., Khosravi-Darani K., Mohammadifar M. A., Nikoopour, H., Production of Mycoprotein by Fusarium Venenatum Growth on Date Sugar, Asian Journal of Chemistry, 21(5), p. 4017 (2009).

[22] Khosravi-Darani K., Zoghi A., Comparison of Pretreatment Strategies of Sugarcane Baggase: Experimental Design for Citric Acid Production, Bioresource Technology, 99, p. 6986 (2008).

[23] Mokhtari-Hosseini Z.B., Vasheghani-Farahani E., Heidarzadeh-Vazifekhoran A., Shojaosadati S.A., Karimzadeh R., Khosravi-Daran K., Statistical Media Optimization for Growth and PHB Production from Methanol by a Methylotrophic Bacterium, Bioresour. Technol., 100, p. 2436 (2009).

[24] Ghaemi-Oskouie S.F., Tabandeh F., Yakhchali B., Eftekhar F., Response Surface Optimization of Medium Composition for Alkaline Protease Production by Bacillus clausii, Biochemical Engineering Journal, 39, p. 37 (2008).

[25] Sarrafzadeh M.H., Belloy L., Esteban G., Navarro J.M., Ghommidh C., Dielectric Monitoring of the Growth and Sporulation of Bacillus thuringiensis. Biotechnology Letters, 27, p. 511 (2005).

[26] Arcas J., Yantorno O., Ertola R., Effect of High Concentration of Nutrients on Bacillus thuringiensis Cultures, Biotechnology Letters, 9 (2), p. 105 (1987).

[27] Wan M., Wan Y., Mohd M., Chan-Choy M., Effect of Ammonium Sulphat on the Sporulation of B. thuringiensis (Local Isolate) During Batch Fermentation., J. Tech., 39, p. 53 (2003).

[28] Zeinat K.M., Nashwa A.H., Fetyan A., Mohamed A.I., Sherif E., Biodegradation and Detoxification of Malathion by B. thuringiensis MOS-5, Australian Journal of Basic and Applied Sciences, 2(3), p. 724, ISSN 1991-8178 (2008).