Effects of Cultural Conditions on Exopolysaccharide Production by Bacillus sp. ZBP4

Microbial exopolysaccharides (EPSs) are of great interest for the application in various industries due to their gelling, stabilizing, emulsifying, and antioxidant properties. In the present study, EPS production of 12 Bacillus strains were investigated and the best producer, namely Bacillus sp. ZBP4, was selected for further studies in order to determine the effects of fermentation conditions on the biosynthesis of EPSs. Beet molasses was used as substrate in the experiments. The highest amount of EPS was obtained at 60 g L-1 molasses concentration within 24 h. Optimum temperature and pH were determined as 45 oC and 5.0, respectively. Various carbon sources (glucose, starch, lactose, whey, mannitol, sucrose, beet molasses) have been tested for EPS production and beet molasses was found as the best. Using inorganic nitrogen source (ammonium sulfate) caused a decrease in the production of EPS. Tryptone gave the highest EPS yields amongst the organic nitrogen sources (yeast extract, peptone, tryptone) tested. Considerable increase in EPS production (1071 mg L-1) has been observed when the experiment was conducted under the optimized conditions (using tryptone and 60 g L-1 molasses at pH 5.0 and 45 °C in 24 h) which was 143 mg L-1 before the optimization studies.

Keywords:

Bacillus; Exopolysaccharide (EPS); Beet molasses,

PDF

___

Abdel-Aziz S M, Hamed H A, Mouafi F E & Gad A S (2012). Acidic pH-shock induces the production of an exopolysaccharide by the fungus Mucor rouxii: Utilization of beet-molasses. New York Science Journal 5(2): 52-61
Abdul Razack S, Velayutham V & Thangavelu V (2013). Medium optimization for the production of exopolysaccharide by Bacillus subtilis using synthetic sources and agro wastes. Turkish Journal of Biology 37: 280-288
Avcı A, Cagri-Mehmetoglu A & Arslan D (2017). Production of antimicrobial substances by a novel Bacillus strain inhibiting Salmonella Typhimurium. LWT Food Science and Technology 80: 265-270
Castillo E & Lopez-Mungia A (2004). Synthesis of levan in water-miscible organic solvents. Journal of Biotechnology 114(1-2): 209-217
Çelik G Y, Aslım B & Beyatli Y (2008). Characterization and production of the exopolysaccharide (EPS) from Pseudomonas aeruginosa G1 and Pseudomonas putida G12 strains. Carbohydrate Polymers 73: 178-182
De Vuyst L & Degeest B (1999). Heteropolysaccharides from lactic acid bacteria. FEMS Microbiology Reviews 23: 153-177
Donot F, Fontana A, Baccou J C & Galindo S S (2012). Microbial exopolysaccharides: main examples of synthesis, excretion, genetics and extraction. Carbohydrate Polymers 87: 951-962
Dubois M, Gilles K A, Hamilton J K, Rebers P A & Smith F (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28: 350-356
Fang Y, Ahmed S, Liu S, Wang S, Lu M & Jiao Y (2013). Optimization of antioxidant exopolysaccharides production by Bacillus licheniformis in solid state fermentation. Carbohydrate Polymers 98: 1377-1382
Freitas F, Alves V D & Reis M A M (2011). Advances in bacterial exopolysaccharides: From production to biotechnological applications. Trends in Biotechnology 29: 388-398
Göksungur Y, Uçan A & Güvenç U (2004). Production of pullulan from beet molasses and synthetic medium by Aureobasidium pullulans. Turkish Journal of Biology 28: 23-30
Koçberber K N & Dönmez G (2008). Environmental conditions affecting exopolysaccharide production by Pseudomonas aeruginosa, Micrococcus sp. and Ochrobactrum sp. Journal of Hazardous Materials 154: 1019-1024
Kumar T (2012). Microbial extracellular polymeric substances production, isolation and applications. IOSR Journal of Pharmacy 2(2): 276-281
Larpin S, Sauvageot N, Pichereau V, Laplace J M & Auffray Y (2002). Biosynthesis of exopolysaccharide by a Bacillus licheniformis strain isolated from ropy cider. International Journal of Food Microbiology 77: 1-9
Li W, Ji J, Chen X, Jiang M, Rui X & Dong M (2014). Structural elucidation and antioxidant activities of exopolysaccharides from Lactobacillus helveticus MB2-1. Carbohydrate Polymers 102: 351-359
Liu J, Luo J, Ye H, Sun Y, Lu Z & Zeng X (2009). Production, characterization and antioxidant activities in vitro of exopolysaccharides from endophytic bacterium Paenibacillus polymyxa EJS-3. Carbohydrate Polymers 78: 275-281
Mata J A, Béjar V, Llamas I, Arias S, Bressollier P, Tallon R, Urdaci M C & Quesada E (2006). Exopolysaccharides produced by the recently described halophilic bacteria Halomonas ventosae and Halomonas anticariensis. Research in Microbiology 157: 827-835
Miller G L (1959). Use of dinitrosalycylic acid reagent for determination of reducing sugar. Analytical Chemistry 31: 662-666 Öztürk S, Aslım B, Suludere B & Tan S (2014). Metal removal of cyanobacterial exopolysaccharides by uronic acid content and monosaccharide composition. Carbohydrate Polymers 101: 265-271
Salehizadeh H & Shojaosadati S A (2003). Removal of metal ions from aqueous solution by polysaccharide produced from Bacillus firmus. Water Research 37(17): 4231-4235
Shih I, Chen L D & Wu J Y (2010). Levan production using Bacillus subtilis natto cells immobilized on alginate. Carbohydrate Polymers 82(1): 111-117
Singh R P, Shukla M K, Mishra A, Kumari P, Reddy C R K & Jha B (2011). Isolation and characterization of exopolysaccharides from seaweed associated bacteria Bacillus licheniformis. Carbohydrate Polymers 84: 1019-1026
Sirajunnisa A, Vijayagopal V & Viruthagiri T (2012). Effect of synthetic carbon substrates and cane molasses, an agro waste, on exopolysaccharide production by P. fluorescens. International Journal of Science and Engineering Applications 1(1): 60-66
Tallon R, Bressollier P & Urdaci M C (2003). Isolation and characterization of two exopolysaccharides produced by Lactobacillus plantarum EP56. Research in Microbiology 154: 705-712
Van Geel-Schutten G H, Flesch F, ten Brick B, Smith M R & Dijkhuizen L (1998). Screening and characterization of Lactobacillus strains producing large amounts of exopolysaccharide. Applied Microbiology and Biotechnology 50: 697-703
Wang C L, Huang T H, Liang T W, Fang C Y, San-Lang & Wang S L (2011). Production and characterization of exopolysaccharides and antioxidant from Paenibacillus sp. TKU023. New Biotechnology 28(6): 559-565
Welman A D, Ian S & Maddox I S (2003). Exopolysaccharides from lactic acid bacteria: Perspectives and challenges. Trends in Biotechnology 21(6): 269-274
Yilmaz M, Yuvali Celik G, Aslim B & Onbasili D (2012). Influence of carbon sources on the production and characterization of the exopolysaccharide (EPS) by Bacillus sphaericus 7055 strain. Journal of Polymers and Environment 20: 152-156
Zhou F, Wu Z, Chen C, Han J, Ai L & Guo B (2014). Exopolysaccharides produced by Rhizobium radiobacter S10 in whey and their rheological properties. Food Hydrocolloids 36: 362-368