Gellan Gum Production Research Articles

Optimization of culture medium compositions for gellan gum production by a halobacterium Sphingomonas paucimobilis

The effect of culture medium compositions on gellan gum production produced by fermentation with a halobacterium Sphingomonas paucimobilis QHZJUJW CGMCC2428 was studied. In this work, a fractional factorial design was applied to investigate the main factors that affected gellan gum production by S. paucimobilis QHZJUJW CGMCC2428. Sucrose was the best carbon source for gellan gum and peptone displayed better inducing effect. Central composite design and response surface methodology were adopted to derive a statistical model for optimizing submerged culture medium composition. These experimental results showed that the optimum culture medium for producing gellan gum was composed of 40.00 (w/v) sucrose, 3.00% peptone (w/v), MgSO4 (w/v), 9.20% KH2PO4 (w/v), 7.50% Na2HPO4 (w/v), 4.30% K2SO4 (w/v), pH 6.8–7.0. The maximal gellan gum was 19.89±0.68g/L, which was agreed closely with the predicated value (20.12g/L). After incubated for 72h under the optimized culture medium in 5-L bioreactor, the gellan gum fermentation reached about 19.90±0.68g/L, which was higher than that in the initial cultivation medium.

Enhanced gellan gum production by hydrogen peroxide (H2O2) induced oxidative stresses in Sphingomonas paucimobilis

In this study, the effect of H2O2-induced oxidative stress on gellan gum production and cell growth were investigated. Gellan gum production was improved and cell growth was inhibited by H2O2. A multiple H2O2 stresses with different concentrations were developed to optimize gellan gum production. A maximal gellan gum yield (22.52g/L), which was 35.58% higher than the control, was observed with 2, 2, 3, 4mmol/L H2O2 added at 6, 12, 18, 24h, respectively. Moreover, UDP-glucose pyrophosphorylase activity and glucosyltransferase activity were increased with H2O2 stresses. This new strategy of multiple H2O2-induced oxidative stresses would be further applied to gellan gum production in future study.

Effects of carbon/nitrogen ratio, dissolved oxygen and impeller type on gellan gum production in Sphingomonas paucimobilis

Among exopolysaccharides, gellan gum is used widely as a gelling agent and food additive, and its commercial market is rapidly increasing. To optimize the fermentation production of gellan gum in Sphingomonas paucimobilis ZJU3108, carbon/nitrogen (C/N) ratio, dissolved oxygen (DO) and impeller type were studied. The C/N ratio, rather than the absolute concentration of carbon or nitrogen source, was found to be the major factor affecting cell growth. A C/N ratio above 20.7 had no significant effect on gellan gum production. Although the final yield of gellan gum increased with C/N ratio increase, the glucose to gellan gum conversion rate decreased. This was due to the high viscosity, which caused oxygen limitation from 16 h to 40 h in the inhomogeneous broth. Due to low stationary phase oxygen demand, the effect of DO on biomass and gellan gum production was reduced. A simple DO control strategy was developed to maintain 30% DO for the first 32 h cultivation, followed by anaerobic conditions for the remaining 8 h. Comparison of different impellers suggested that an anchor impeller with impeller diameter of 11.0 cm (D* = 11.0 cm) was suitable for high yields of high molecular weight gellan gum. These studies simplify production of gellan gum by fermentation and improve its yield.

Cloning and knockout of phytoene desaturase gene in Sphingomonas elodea ATCC 31461 for economic recovery of gellan gum

A gene encoding phytoene desaturase (crtI) in the carotenoid biosynthetic pathway of Sphingomonas elodea ATCC 31461, an industrial gellan gum-producing strain, was cloned and identified. This gene is predicted to encode a 492-amino acid protein with significant homology to the phytoene desaturase of other carotenogenic organisms. Knockout of crtI gene blocked yellow carotenoid pigment synthesis and resulted in the accumulation of colorless phytoene, confirming that it encodes phytoene desaturase. Further research indicates that the yield of gellan gum production by crtI gene knockout mutants is almost the same as that by the wild-type strain. In addition, a recovery method based on the colorless fermentation broth of the crtI gene knockout mutant was investigated. Compared to the volume of alcohol for the parent strain, much less alcohol (30%) is required in this recovery process; thus, the costs of downstream purification of gellan gum can be substantially reduced.

Optimization of gellan gum production by Sphingomonas paucimobilis ATCC 31461 with nonionic surfactants using central composite design

The effect of nonionic surfactants on gellan production by Sphingomonas paucimobilis was studied by the addition of 0.5, 0.75, 1.0, 1.25 and 1.5 g/l surfactants to shake flask culture. The nonionic surfactants Tween 80, Tween 40 and Triton X-100 improved gellan production by S. paucimobilis, and the maximum yield (10.44 g/l) was obtained with Triton X-100 at 0.75 g/l compared with that of the control fermentation (8.63 g/l) without surfactant. The DO profiles associated with gellan production in a 5-l laboratory fermentor showed higher oxygen and mass transfers during fermentation with surfactant than during control fermentation without surfactant. The addition of surfactant also resulted in a polymer with high viscosity as manifested by its lower acetyl content, than that obtained by control fermentation. A central composite design (CCD) was used to determine the maximum gellan production at optimum values for three process parameters (Triton X-100 concentration, pH, and temperature) each at five levels in a laboratory fermentor. The maximum gellan yield (14.62 g/l) was obtained in a 5-l laboratory fermentor with 1.0 g/l Triton X-100 and at pH 6.0 and temperature 29.6 degrees C. Further studies on the effects of agitation and DOT level demonstrated that the surfactants enhanced oxygen transfer resulting in higher gellan production (27.86 g/l) at higher agitation speed (1000 rpm) and 100% DOT level.

Statistical approach to optimization of fermentative production of gellan gum from Sphingomonas paucimobilis ATCC 31461

Gellan gum, a high-molecular-weight anionic linear polysaccharide produced by pure-culture fermentation from Sphingomonas paucimobilis ATCC 31461, has elicited industrial interest in recent years as a high-viscosity biogum, a suspending agent, a gelling agent, and an agar substitute in microbial media. In this paper we report on the optimization of gellan gum production using a statistical approach. In the first step, the one factor-at-a-time method was used to investigate the effect of medium constituents such as carbon and nitrogen sources; subsequently, the intuitive analysis based on statistical calculations carried out using the L16 -orthogonal array method. The design for the L16 -orthogonal array was developed and analyzed using MINITAB 13.30 software. All the fermentation runs were carried out at 30+/-2 degrees C on a rotary orbital shaker at 180 rpm for 48 h. In the second step, the effects of amino acids and gellan precursors such as uridine-5'-diphospate (UDP) and adenosine-5'-diphospate (ADP) on the fermentative production of gellan gum were studied. Media containing 4% soluble starch, 0.025% yeast extract, 1.0 mM ADP and 0.05% tryptophan gave a maximum yield of 43.6 g l(-1) starch-free gellan gum, which was significantly higher than reported values in the literature.

Improvement in Production and Quality of Gellan Gum by Sphingomonas paucimobilis Under High Dissolved Oxygen Tension Levels

The effect of agitation rate and dissolved oxygen tension (DOT) on growth and gellan production by Sphingomonas paucimobilis was studied. Higher cell growth of 5.4 g l(-1) was obtained at 700 rpm but maximum gellan (15 g l(-1)) was produced at 500 rpm. DOT levels above 20% had no effect on cell growth but gellan yield was increased to 23 g l(-1 )with increase in DOT level to 100%. Higher DOT levels improved the viscosity and molecular weight of the polymer with change in acetate and glycerate content of the polymer.

Optimization of nutrients for gellan gum production by Sphingomonas paucimobilis ATCC-31461 in molasses based medium using response surface methodology

A molasses based medium for the production of gellan by Sphingomonas paucimobilis ATCC-31461 was developed. Placket–Burman design criterion was applied to study the effect of various nutrient supplements on gellan production using molasses. Among the 20 variables tested, molasses, tryptone, casaminoacid, disodium hydrogen orthophosphate and manganese chloride showed significant effect on gellan production. A central composite design was applied to determine the optimum concentrations of the significant variables obtained from Placket–Burman design. Most suitable medium composition for production of gellan was (g/l): molasses—112.5; tryptone—1; casaminoacid—1; disodium hydrogen orthophosphate—1; manganese chloride—0.947 and the optimum gellan production was 13.814 g/l.

Modeling for gellan gum production by Sphingomonas paucimobilis ATCC 31461 in a simplified medium.

Gellan gum production was carried out by Sphingomonas paucimobilis ATCC 31461 in a simplified medium with a short incubation time, and a kinetic model for understanding, controlling, and optimizing the fermentation process was proposed. The results revealed that glucose was the best carbon source and that the optimal concentration was 30 g liter(-1). As for the fermenting parameters, considerably large amounts of gellan gum were yielded by an 8-h-old culture and a 4% inoculum at 200 rpm on a rotary shaker. Under the optimized conditions, the maximum level of gellan gum (14.75 g liter(-1)) and the highest conversion efficiency (49.17%) were obtained in a 30-liter fermentor in batch fermentation. Logistic and Luedeking-Piret models were confirmed to provide a good description of gellan gum fermentation, which gave some support for the study of gellan gum fermentation kinetics. Additionally, this study is the first demonstration that gellan gum production is largely growth associated by analysis of kinetics in its batch fermentation process. Based on model prediction, higher gellan gum production (17.71 g liter(-1)) and higher conversion efficiency (57.12%) were obtained in fed-batch fermentation at the same total glucose concentration (30 g liter(-1)).

Production of gellan gum by Sphingomonas paucimobilis NK2000 with soybean pomace

The ammonium nitrate as a nitrogen source for the production of gellan gum by Sphingomonas paucimobilis NK2000 enhanced the cell growth but its depletion was essential for higher production of gellan gum. The highest production of gellan gum was 3.27 g l −1 when the concentrations of glucose and bacto-peptone were 2.0% (w/v) and 0.05% (w/v). The production of gellan gum by S. paucimobilis NK2000 increased with increased concentration of soybean pomace substituted for bacto-peptone up to 2.0% (w/v). The highest production of gellan gum was 7.33 g l −1 in a flask scale when the concentrations of glucose and soybean pomace without ammonium nitrate were 2.0% and 2.0% (w/v). The highest production of gellan gum with the same concentrations of glucose and soybean pomace was 7.50 g l −1 in a 7 l bioreactor whereas that with 2.0% (w/v) glucose and 0.05% (w/v) bacto-peptone was 4.95 g l −1.

Effect of environmental factors and carbohydrate on gellan gum production.

Submerged culture fermentation studies were carried out in batch mode for optimizing the environmental parameters and carbon source requirement by Pseudomonas elodea for the production of gellan gum. The maximum production of gellan gum was obtained with 16-h-old culture and 8% inoculum at 30 degrees C and pH 7.0 after 52 h of incubation (6.0 g/L). Of the various carbon sources tested, 2% sucrose, glucose, and soluble starch yielded considerably high amounts of gellan. Studies on the concentration of various carbohydrates on gellan gum production indicated that the optimum concentration of glucose and starch was 3%, whereas for sucrose it was 4%. The addition of glucose in the medium above 3% had a detrimental effect on gellan yield. The investigation of intermediate two-step addition of glucose under identical conditions of fermentation showed an enhanced production of gellan (8.12 g/L) as compared with the control (6.0 g/L). To optimize the recovery of gellan from fermented broth, different solvents were tested for precipitation of gellan gum. Among the various solvents tested, tetrahydrofuran gave better recovery of gellan (82%) as compared with the conventional solvent isopropanol (49%).

Exopolysaccharide of the gellan family: prospects and potential

The use of microbial polysaccharides in the food, pharmaceutical and chemical industries has increased steadily during the past decade. The biopolymer gellan is a more recent addition to the family of microbial polysaccharides that is gaining much importance due to its novel property of forming thermo-reversible gels when heated and cooled. It is produced and marketed by some companies of Europe, USA, etc under trade names such as ‘Gelrite’, ‘Phytagel’ and ‘Kelcogel’. It has applications in diverse fields in the food, pharmaceutical and many other industries. Further research and development in biopolymer technology is expected to expand its use. This article presents a critical review of the available published information on the gellan exopolysaccharide synthesized by Pseudomonas species. In particular information on its structure, physico-chemical properties and the rheology of its solutions etc. is critically assessed. Emphasis has also been paid to characterization of gellan. A brief historical background of the polymer and the biochemical and physiological characteristics of several different existing bacterial isolates which secrete gellan and related polysaccharides are discussed. An attempt has also been made to review the potential and future prospects, highlighting some novel techniques adopted to overcome the mass transfer problems associated with the fermentative production of gellan gum. The efficient downstream processes used for obtaining purified gellan are also highlighted. Attention has also been drawn to the problem associated with the fermentation processes due to the highly viscous nature of gellan gum and effect of different impeller systems on gellan fermentation kinetics and rheological properties.