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Statistical approach to optimize production of biosurfactant by Pseudomonas aeruginosa 2297.

ABSTRACT: The main objective of this paper is to optimize biosurfactant production by Pseudomonas aeruginosa 2297 with statistical approaches. Biosurfactant production from P. aeruginosa 2297 was carried out with different carbon sources, and maximum yield was achieved with sawdust followed by groundnut husk and glycerol. The produced biosurfactant has showed active emulsification and surface-active properties. From the kinetic growth modeling, the specific growth rate was calculated on sawdust and it was 1.12 day^-1. The maximum estimated value of product yield on biomass growth (Y_p/x) was 1.02 g/g. The important medium components identified by the Plackett-Burman method were sawdust and glycerol along with culture parameter pH. Box-Behnken response surface methodology was applied to optimize biosurfactant production. The obtained experimental result concludes that Box-Behnken designs are very effective statistical tools to improve biosurfactant production. These results may be useful to develop a high efficient production process of biosurfactant. In addition, this type of kinetic modeling approach may constitute a useful tool to design and scaling-up of bioreactors for the production of biosurfactant.

SUBMITTER: Kumar AP

PROVIDER: S-EPMC4327757 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Statistical approach to optimize production of biosurfactant by Pseudomonas aeruginosa 2297.

Kumar Arthala Praveen AP Janardhan Avilala A Radha Seela S Viswanath Buddolla B Narasimha Golla G

3 Biotech 20140308 1

The main objective of this paper is to optimize biosurfactant production by Pseudomonas aeruginosa 2297 with statistical approaches. Biosurfactant production from P. aeruginosa 2297 was carried out with different carbon sources, and maximum yield was achieved with sawdust followed by groundnut husk and glycerol. The produced biosurfactant has showed active emulsification and surface-active properties. From the kinetic growth modeling, the specific growth rate was calculated on sawdust and it was ...[more]

PMID: 28324363

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Project description:Tea is an ancient non-alcoholic beverage plantation crop cultivated in the most part of Assam, India. Being a long-term monoculture, tea plants are prone to both biotic and abiotic stresses, and requires massive amounts of chemicals as fertilizers and pesticides to achieve worthy crop productivity. The rhizosphere bacteria with the abilities to produce phytohormone, secreting hydrolytic enzyme, biofilm formation, bio-control activity provides induced systemic resistance to plants against pathogens. Thus, plant growth promoting (PGP) rhizobacteria represents as an alternative candidate to chemical inputs for agriculture sector. Further, deciphering the secondary metabolites, including biosurfactant (BS) allow developing a better understanding of rhizobacterial strains. The acidic nature of tea rhizosphere is predominated by Bacillus followed by Pseudomonas that enhances crop biomass and yield through accelerating uptake of nutrients. In the present study, a strain Pseudomonas aeruginosa RTE4 isolated from tea rhizosphere soil collected from Rosekandy Tea Garden, Cachar, Assam was evaluated for various plant-growth promoting attributes. The strain RTE4 produces indole acetic acid (74.54 μg/ml), hydrolytic enzymes, and solubilize tri-calcium phosphate (46 μg/ml). Bio-control activity of RTE4 was recorded against two foliar fungal pathogens of tea (Corticium invisium and Fusarium solani) and a bacterial plant pathogen (Xanthomonas campestris). The strain RTE4 was positive for BS production in the preliminary screening. Detailed analytical characterization through TLC, FTIR, NMR, and LCMS techniques revealed that the strain RTE4 grown in M9 medium with glucose (2% w/v) produce di-rhamnolipid BS. This BS reduced surface tension of phosphate buffer saline from 71 to 31 mN/m with a critical micelle concentration of 80 mg/L. Purified BS of RTE4 showed minimum inhibitory concentration of 5, 10, and 20 mg/ml against X. campestris, F. solani and C. invisium, respectively. Capability of RTE4 BS to be employed as a biofungicide as compared to Carbendazim - commercially available fungicide is also tested. The strain RTE4 exhibits multiple PGP attributes along with production of di-rhamnolipid BS. This gives a possibility to produce di-rhamnolipid BS from RTE4 in large scale and explore its applications in fields as a biological alternative to chemical fertilizer.

Project description:BACKGROUND:Rhamnolipids are the most extensively studied biosurfactants and has been successfully used in various areas from bioremediation to industrial fields. Rhamnolipids structural composition decide their physicochemical properties. Different physicochemical properties influence their application potential. Rhamnolipids can be produced at both aerobic conditions and anaerobic conditions by Pseudomonas aeruginosa. This study aims to evaluate the oxygen effects on the rhamnolipids yield, structural composition, physicochemical properties and the rhl-genes expression in P. aeruginosa SG. Results will guide researchers to regulate microbial cells to synthesize rhamnolipids with different activity according to diverse application requirements. RESULTS:Quantitative real-time PCR analysis revealed that rhlAB genes were down-regulated under anaerobic conditions. Therefore, strain P. aeruginosa SG anaerobically produced less rhamnolipids (0.68 g/L) than that (11.65 g/L) under aerobic conditions when grown in media containing glycerol and nitrate. HPLC-MS analysis showed that aerobically produced rhamnolipids mainly contained Rha-C8-C10, Rha-Rha-C10-C12:1 and Rha-Rha-C8-C10; anaerobically produced rhamnolipids mainly contained Rha-C10-C12 and Rha-C10-C10. Anaerobically produced rhamnolipids contained more mono-rhamnolipids (94.7%) than that (54.8%) in aerobically produced rhamnolipids. rhlC gene was also down-regulated under anaerobic conditions, catalyzing less mono-rhamnolipids to form di-rhamnolipids. Aerobically produced rhamnolipids decreased air-water surface tension (ST) from 72.2 to 27.9 mN/m with critical micelle concentration (CMC) of 60 mg/L; anaerobically produced rhamnolipids reduced ST to 33.1 mN/m with CMC of 80 mg/L. Anaerobically produced rhamnolipids emulsified crude oil with EI24 = 80.3%, and aerobically produced rhamnolipids emulsified crude oil with EI24 = 62.3%. Both two rhamnolipids products retained surface activity (ST < 35.0 mN/m) and emulsifying activity (EI24 > 60.0%) under temperatures (4-121 °C), pH values (4-10) and NaCl concentrations less than 90 g/L. CONCLUSIONS:Oxygen affected the rhl-genes expression in P. aeruginosa, thus altering the rhamnolipids yield, structural composition and physicochemical properties. Rhamnolipids produced at aerobic or anaerobic conditions was structurally distinct. Two rhamnolipids products had different application potential in diverse biotechnologies. Although both rhamnolipids products were thermo-stable and halo-tolerant, aerobically produced rhamnolipids possessed better surface activity, implying its well wetting activity and desorption property; anaerobically produced rhamnolipids exhibited better emulsifying activity, indicating its applicability for enhanced oil recovery and bioremediation of petroleum pollution.

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Statistical approach to optimize production of biosurfactant by Pseudomonas aeruginosa 2297.

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Statistical approach to optimize production of biosurfactant by Pseudomonas aeruginosa 2297.

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