Statistical Design, a Powerful Tool for Optimizing Biosurfactant Production: A Review <s> Strains, Bs Classification and Metabolism <s> The production of biosurfactant from Rhodococcus spp. MTCC 2574 was effectively enhanced by response surface methodology (RSM). Rhodococcus spp. MTCC 2574 was selected through screening of seven different Rhodococcus strains. The preliminary screening experiments (one-factor at a time) suggested that carbon source: mannitol, nitrogen source: yeast extract and meat peptone and inducer: n-hexadecane are the critical medium components. The concentrations of these four media components were optimized by using central composite rotatable design (CCRD) of RSM. The adequately high R2 value (0.947) and F score 19.11 indicated the statistical significance of the model. The optimum medium composition for biosurfactant production was found to contain mannitol (1.6 g/L), yeast extract (6.92 g/L), meat peptone (19.65 g/L), n-hexadecane (63.8 g/L). The crude biosurfactant was obtained from methyl tert-butyl ether extraction. The yield of biosurfactant before and after optimization was 3.2 g/L of and 10.9 g/L, respectively. Thus, RSM has increased the yield of biosurfactant to 3.4-fold. The crude biosurfactant decreased the surface tension of water from 72 mN/m to 30.8 mN/m (at 120 mg L(-1)) and achieved a critical micelle concentration (CMC) value of 120 mg L(-1). <s> BIB001 </s> Statistical Design, a Powerful Tool for Optimizing Biosurfactant Production: A Review <s> Strains, Bs Classification and Metabolism <s> Microbial biosurfactants are surface active metabolites synthesized by microbes growing on a variety of substrates. In spite of having great potential for commercial, therapeutic and environmental applications, industrial level production has not been realized for their low yields and productivities. One vital factor determining their biosynthesis is the genetic makeup of the producer organisms. Studies on molecular genetics and biochemistry of the synthesis of several biosurfactants have revealed the operons, the enzymes and the metabolic pathways required for their extracellular production. Surfactin, a cyclic lipopeptide biosurfactant is a potent antimicrobial agent and is produced as a result of non-ribosomal biosynthesis catalyzed by a large multienzyme peptide synthetase complex called the surfactin synthetase. Pathways for the synthesis of other lipopeptides such as iturin, lichenysin and arthrofactin are also mediated by similar enzyme complexes. These non-ribosomal peptide synthetases (NRPSs) responsible for lipopeptide biosynthesis display a high degree of structural similarity among themselves even from distant microbial species. Plasmid-encoded- rhlA, B, R and I genes of rhl quorum sensing system are required for production of glycolipid biosurfactants by Pseudomonas species. Molecular genetics of biosynthesis of alasan and emulsan by Acinetobacter species and of the fungal biosurfactants such as mannosylerythritol lipids (MEL) and hydrophobins have been deciphered. However, limited genetic information is available about biosynthesis of other biosurfactants such as viscosin, amphisin and putisolvin produced by some strains of Pseudomonas species. Understanding of the genetic regulatory mechanisms would help to develop metabolically engineered hyper-producing strains with better product characteristics and acquired capability of utilizing cheap agro-industrial wastes as substrates. This article thus provides an overview of the role and importance of molecular genetics and gene regulation mechanisms behind the biosynthesis of various microbial surfactants of commercial importance. <s> BIB002 </s> Statistical Design, a Powerful Tool for Optimizing Biosurfactant Production: A Review <s> Strains, Bs Classification and Metabolism <s> The strain SmSA, identified as Serratia marcescens and known as a biosurfactant producer, was isolated from hydrocarbon contaminated soil from Veracruz, México. The interactions among the C/N, C/Mg and C/Fe ratios have not been examined for this microorganism. In this work was evaluated the effect of these nutrients at three levels using a mineral medium with glucose as the carbon source. A Box-Behnken experimental design was utilised to maximise biosurfactant production, which was assessed by oil spreading and surface tension tests. The treatment with C/N=5, C/Fe=26,000 and C/Mg=30 showed the best result since the surface tension was reduced to 30 mN m(-1). The multiple regression and response surface analyses indicated that the interaction between C/N and C/Mg had the utmost effect on the reduction of surface tension and biosurfactant production. The conditions of the best treatment were used to scale up biosurfactant production in a 3L bioreactor giving a yield of 4.1 gL(-1) of pure biosurfactant. It was found that the biosurfactant was mainly produced in the exponential phase and decreased the surface tension to 31 mN m(-1). The contact between the biosurfactant with heavy oil (15° API) increased its displacement from 9.3 to 18 cm. <s> BIB003
There are many microorganisms that have been reported to produce Bs including fungi, yeast, and bacteria, although bacteria are the most exploited. Pseudomonas and Bacillus are generally described as super-producers  . Other genera such as Serratia, Vibrio, Rhodococcus, Strepococcus, Acinetobacter, and even Lactococcus have also been reported to produce good amounts of Bs  BIB003 BIB001 . Additionally, yeast such as Candida, Yarrowia, Saccharomyces, and Kluveromyces have been extensively studied for their Bs producing capacities. The Bs produced by different microorganisms are classified into glycolipids, lipopeptides, fatty acids, phospholipids, polymeric surfactants, and particulate surfactants  BIB002 . Each group of Bs can be further divided into sub classifications. For example, glycolipids can be divided into rhamnolipids, trehalolipids, or sophorolipids. Glycolipids are generally produced by a wide range of bacteria and yeast, while other types of surfactants are reduced to a more limited range of bacteria. For example, Bacillus is generally prone to synthesize lipopeptide Bs and do not produce glycolipids. Acinetobacter and Pseudomonas are known to produce a diversity of Bs  BIB002 .