Gellan Production Research Articles

Characterization and optimization of gellan gum production by natural Sphingomonas sp. SM2

Gellan is one of the most important industrial microbial exopolysaccharides used for a wide range of applications in food, pharmaceutical, and oil. In this study, a native strain of Sphingomonas producing gellan gum was identified from rice roots. Various factors, including carbon and nitrogen sources, pH levels, fermentation duration, and aeration, were investigated to enhance the production of gellan by Sphingomonas sp. SM2. The highest efficiency in gum production was achieved by a medium composed of 30 g/L glucose. When the carbon-to-nitrogen ratio was 30, organic nitrogen caused this increase. By incorporating all relevant factors, the optimized fermentation process can yield a gum concentration of 8.64 ± 0.34 g/L. The total carbohydrate content of the native gum was found to be 52.62 ± 0.52%. The existence of mucilage surrounding the bacterial cells was verified by scanning electron microscopy images. The extracted gellan gum was characterized using FTIR, 1H NMR, and HPLC analyses. The structure of the gum was confirmed by comparing it with the standard gellan gum. The production of gellan gum by Sphingomonas sp. SM2 using the utilization of diverse carbon sources suggests its immense potential as a gellan producer.

Exploring Potential of Gellan Gum for Enhanced Oil Recovery.

Extensive laboratory and field tests have shown that the gelation response of gellan gum to saline water makes it a promising candidate for enhanced oil recovery (EOR). The objective of this mini-review is to evaluate the applicability of gellan gum in EOR and compare its efficiency to other precursors, in particular, hydrolyzed polyacrylamide (HPAM). At first, the "sol-gel" phase transitions of gellan gum in aqueous-salt solutions containing mono- and divalent cations are considered. Then the rheological and mechanical properties of gellan in diluted aqueous solutions and gel state are outlined. The main attention is paid to laboratory core flooding and field pilot tests. The plugging behavior of gellan in laboratory conditions due to "sol-gel" phase transition is discussed in the context of conformance control and water shut-off. Due to its higher strength, gellan gum gel provided ~6 times greater resistance to the flow of brine in a 1 mm-width fracture compared to HPAM gel. The field trials carried out in the injection and production wells of the Kumkol oilfield, situated in Kazakhstan, demonstrated that over 6 and 11 months, there was an incremental oil recovery of 3790 and 5890 tons, respectively. To put it into perspective, using 1 kg of dry gellan resulted in the incremental production of 3.52 m3 (or 22 bbls) of oil. The treatment of the production well with 1 wt.% gellan solution resulted in a considerable decrease in the water cut up to 10-20% without affecting the oil flow rate. The advantages and disadvantages of gellan compared to HPAM are analyzed together with the economic feasibility of gellan over HPAM. The potential for establishing gellan production in Kazakhstan is emphasized. It is anticipated that gellan gum, manufactured through fermentation using glucose-fructose syrup from Zharkent and Burunday corn starch plants, could be expanded in the future for applications in both the food industry and oil recovery.

The fermentation-based production of gellan from rice bran and the evaluation of various qualitative properties of gum

The potential for the use of rice bran, an agricultural waste, as a substrate in the manufacture of gellan gum was examined. Using a standard strain of Sphingomonas paucimobilis (ATCC 31461) and rice bran substrate, gellan gum was produced under optimized conditions. The optimal yield of gellan gum using rice bran substrate was found to be 11.96 g L−1 with 5% glucose, 10% inoculum, and a mixing speed of 300 rpm. Native gum was found to have a consistency index of 2.00 Pa.sn. The viscosity of the gum was found to be extremely stable when exposed to thermal stress. Concerning the rheological characteristics, the Herschel–Bulkley model offered a more realistic representation of the flow characteristics of gum solutions. The synthesized gums were mostly composed of glucose, rhamnose, and glucuronic acid. The acetic acid content of gellan gums was 2.95%, while the molecular weight was 2.88 × 105 Da. Characterization of native gellan gums by UV–Vis spectroscopy, SEM, TEM and FTIR spectroscopy is also presented.

Synthesis of the Microbial Polysaccharide Gellan from Dairy and Plant-Based Processing Coproducts

This review examines the production of the microbial polysaccharide gellan, synthesized by Sphingomonas elodea, on dairy and plant-based processing coproducts. Gellan is a water-soluble gum that structurally exists as a tetrasaccharide comprised of 20% glucuronic acid, 60% glucose and 20% rhamnose, for which various food, non-food and biomedical applications have been reported. A number of carbon and nitrogen sources have been tested to determine whether they can support bacterial gellan production, with several studies attempting to optimize gellan production by varying the culture conditions. The genetics of the biosynthesis of gellan has been explored in a number of investigations and specific genes have been identified that encode the enzymes responsible for the synthesis of this polysaccharide. Genetic mutants exhibiting overproduction of gellan have also been identified and characterized. Several dairy and plant-based processing coproducts have been screened to learn whether they can support the production of gellan in an attempt to lower the cost of synthesizing the microbial polysaccharide. Of the processing coproducts explored, soluble starch as a carbon source supported the highest gellan production by S. elodea grown at 30 °C. The corn processing coproducts corn steep liquor or condensed distillers solubles appear to be effective nitrogen sources for gellan production. It was concluded that further research on producing gellan using a combination of processing coproducts could be an effective solution in lowering its overall production costs.

A Carotenoid- and Poly-β-Hydroxybutyrate-Free Mutant Strain of Sphingomonas elodea ATCC 31461 for the Commercial Production of Gellan.

Gellan gum is a microbial exopolysaccharide, produced after aerobic fermentation using the Gram-negative bacterium strain Sphingomonas elodea ATCC 31461. Due to its unique structure and excellent physical characteristics, gellan gum has a broad range of applications in food, pharmaceutical, and other industries where it is used for stabilizing, emulsifying, thickening, and suspending. During the fermentative production of gellan, strain ATCC 31461 also accumulates large amounts of the metabolic by-products yellow carotenoid pigments and poly-β-hydroxybutyrate (PHB), which is decreasing the gellan production and increasing processing costs. A pigment PHB-free mutant was obtained by knocking out the phytoene desaturase gene (crtI) in the carotenoid biosynthetic pathway and the phaC gene, encoding a PHB synthase for the polymerization of PHB. Unfortunately, the double gene knockout mutant produced only 0.56 g liter-1 gellan. Furthermore, blocking PHB and carotenoid synthesis resulted in the accumulation of pyruvate, which reduced gellan production. To elevate gellan production, combined UV irradiation and ethyl methanesulfonate (EMS) mutagenesis treatment were used. A mutant strain with the same level of pyruvate as that of the wild-type strain and higher gellan production was isolated (1.35 g liter-1, 132.8% higher than the double gene knockout mutant and 14.4% higher than the wild-type strain ATCC 31461). In addition, a new gellan gum recovery method based on the new mutant strain was investigated, in which only 30% isopropanol was required, which is twice for the wild-type strains, and the performance of the final product was improved. Thus, the mutant strain could be an ideal strain for the commercial production of gellan.IMPORTANCE A carotenoid- and PHB-free double gene knockout strain mutant was constructed to simplify the purification steps normally involved in gellan production. However, the production of gellan gum was unexpectedly reduced. A mutant with 14.4% higher gellan production than that of the wild-type strain was obtained and isolated after employing UV and EMS combined mutagenesis. Based on this high-yield and low-impurity-producing mutant, a new recovery method requiring less organic solvent and fewer operating steps was developed. This method will effectively reduce the production costs and improve the economic benefits of large-scale gellan production.

Effect of 2-deoxy-D-glucose on gellan gum biosynthesis by Sphingomonas paucimobilis.

2-Deoxy-D-glucose (2-DG) is a non-metabolizable glucose analogue and competitive inhibitor of glycolysis. Effect of 2-DG on gellan gum biosynthesis by Sphingomonas paucimobilis ATCC31461 were studied in this research. The concentrationand the addition time of 2-DG significantly affected the biomass and gellan gum accumulation. The maximum gellan gum yield of 20.78g/L was obtained with the addition of 50µg/L of 2-DG at 24h. The mechanism of 2-DG addition favoring to gellan production was revealed by determining the activities of key enzymes. Results indicated that 2-DG addition increased the activities of glucosyltransferase and inhibited UDP-glucose pyrophosphorylase activity. The result indicated that 2-DG inhibited glycolysis and changed metabolic driving force to activate gellan gum biosynthesis metabolism pathways.

Production of Gellan Gum by Sphingomonas paucimobilis on Crude Sweet Whey Using Different Bioreactor Feeding Strategies

Aims: Production of gellan gum by Sphingomoas paucimobilis from whey was optimized by different fermentation techniques.
 Study Design: Study the growth behavior of Sphingomoas paucimobilis was cultivated on 40% sweet whey medium in the bioreactor as a batch, fed batch and continuous culture and effect of aeration and agitation speed on gellan production.
 Place and Duration of Study: Microbiology Dept., Fac. of Agriculture, Ain Shams Univ., Cairo, Egypt, 2016/ 2017.
 Methodology: Using Sphingomoas paucimobilis on sweet whey in the bioreactor as a batch, fed batch (pulsed & continuous) and continuous culture. Among the four levels of air saturation and four levels of agitation speeds.
 Results: Using the continuous feeding of sugar sweet whey at 1.53 gl-1h-1during 12 h was favorable than pulsed feeding for gellan production in fed-batch culture. In continuous culture addition of 40% SW at 0.055 h-1 dilution rate (110 ml h-1), the values of gellan parameters recorded by Sphingomoas paucimobilis were 24.34%, 26.54% & 0.337 gl-1h-1 for gellan yield, conversion coefficient and gellan productivity during 24 h. At 28ºC. This technique increments the gellan production (gl-1h-1) by 3.3 & 2.2-2.5 and 1.5- 1.6 fold as compared to that produced by using batch & fed- batch pulsed and fed-batch continuous techniques respectively. The emulsifying capability of the partially purified gellan was 100% whereas it was 95% for xanthan gum, as well as its high flocculating activity than xanthan. A tough worm-like gel or firm gel were formed when 10% calcium chloride solution or 0.5 g sodium chloride were added to gellan solution.
 Conclusion: The maximum gellan yield and lower fermentation period were obtained with air saturation of 60% at 750 rpm agitation speed. The continuous feeding at 1.53 gl-1h-1 was favorable than pulsed feeding for gellan production in fed-batch culture, while the maximum gellan productivity was obtained by using a continuous culture technique at 0.055 h-1 dilution rate.

Production of gellan gum, an exopolysaccharide, from biodiesel-derived waste glycerol by Sphingomonas spp.

In the present study, biodiesel-derived waste glycerol (WG) was used for the isolation and production of gellan, an exopolysaccharide, on media containing WG as the main carbon source. Two bacterial isolates showed gellan producing potential which were identified as Sphingomonas pseudosanguinis (Accession No. GI:724472387) and Sphingomonas yabuuchiae (GI:724472388) by 16S rRNA gene sequencing. To maximize gellan production by S. pseudosanguinis and S. yabuuchiae, media optimization was performed at different pHs and glycerol concentrations. Morphological observations through microscopic images showed the production of gellan from these isolates. Simple linear regression showed better utilization of WG by S. pseudosanguinis than S. yabuuchiae at pH 6 and pH 7. Though, both the strains showed reverse trend at pH 8. Both the strains were able to produce high amounts of gellan gum (51.6 and 52.6g/l, respectively) using WG (80g/l) as the sole carbon source, in a minimal medium. This is the first report on the efficient degradation of WG and low-cost production of gellan. Owing to these characteristics, S. pseudosanguinis and S. yabuuchiae demonstrate great potential for use in the commercial production of gellan and in the bioremediation of WG.

A new strategy to enhance gellan production by two-stage culture in Sphingomonas paucimobilis

The effects of different initial sucrose concentrations and temperatures on gellan biosynthesis by Sphingomonas paucimobilis ATCC 31461 were investigated. Lower sucrose concentrations and higher temperatures were favorable for cell growth. Higher sucrose concentrations and lower temperatures promoted gellan production but retarded cell growth. Based on these results, a two-stage culture strategy was developed to improve gellan production. During the first 24h, S. paucimobilis was cultured in a pulse fed-batch mode with an initial sucrose concentration 10g/L. Ten grams per liter of sucrose were added at 12h and 24h, and the temperature was controlled at 33°C. Batch culture was performed, and the temperature was reduced to 28°C to achieve a high gellan accumulation. The two-stage culture strategy achieved the highest gellan production (22.61g/L) at 60h that was 35.71% higher than the result of the best conventional batch operation (16.66g/L). Meanwhile, high gellan yield was related to high UDPG-pyrophosphorylase activity and glucosyltransferase activity.

PRODUCTION OF THE MICROBIAL EXOPOLYSACCHARIDE, GELLAN, BY SPHINGOMONAS PAUCIMOBILIS BACTERIA. 2.OPTIMIZATION OF GROWTH CULTURAL COMPOSITION

The purpose of this study was to find the optimum growth cultural composition for the production of the microbial exopolysaccharide, gellan, by the bacteria Sphingomonas paucimobilis using batch cultural method in laboratory media. The study included the effect of the type and concentration of organic compounds as a sole carbon source, type and concentration of organic and non-organic nitrogen source, as well as, the effect of adding of some amino acids and non-ionic surfactants to the gellan production basal medium. The physical cultural conditions were fixed, and the production was carried for 48hrs incubation period, 10% inoculums volume, initial pH of 7.0, incubation temperature at 30oc and agitation speed of 250 rpm in an orbital shaker incubator. Results indicated that the best organic source was glucose at a concentration of 2% as it resulted in 4.44g of gellan/l of production medium, and the best organic nitrogen source was yeast extract at 0.05% and the best non-organic nitrogen source was ammonium chloride at 0.1% concentration, as they gave 4.27 and 4.41 g/l, respectively. On the other hand, the amino acid threonine at 0.05% was the best among other amino acids, and the non-ionic surfactant Triton X-100 at 0.1% was the best and they produced 5029 and 5.59 g of gellan/l of the medium.

A carotenoid-free mutant strain of Sphingomonas paucimobilis ATCC 31461 for the commercial production of gellan

Gellan, an exopolysaccharide produced by Sphingomonas paucimobilis ATCC 31461, is used in the food, pharmaceutical, and other industries. During the fermentative production of gellan, strain ATCC 31461 also accumulates large amounts of yellow carotenoid pigments, thereby increasing processing costs. A carotenoid-free mutant strain of ATCC 31461 was isolated by ethyl methanesulfonate mutagenesis combined with ultraviolet irradiation. A new gellan purification method which reduced the consumption of isopropanol was developed based on this mutant strain. The final gellan production of the mutant strain was 13% higher than that of the parent strain. The glycerate and acetate substituents of gellan produced by the mutant strain were 17% and 65% higher than those produced by gellan from ATCC 31461, respectively. Stability tests suggested that the broth produced by the mutant strain was more stable during storage at room temperature compared with that produced by the parent strain. The mutant strain seems to be an ideal strain for use in the commercial production of gellan.

Constitutive expression of Vitreoscilla haemoglobin in Sphingomonas elodea to improve gellan gum production

To improve a commercially used strain for gellan production by exogenous Vitreoscilla haemoglobin (VHb). VHb gene was expressed in Sphingomonas elodea under the control of constitutive bla promoter. Biochemical activity of expressed VHb was confirmed by CO-difference spectra analysis that exhibited a characteristic absorption maximum at 419 nm. During cultivation, not only enhanced cell growth was detected, but also 20% improvement in gellan production was observed after 48 h of incubation, with a maximum yield of 16·82 g l(-1). Moreover, maximum sucrose conversion efficiency (g gellan per g sucrose) was 57·8, 20% higher than that of the parental strain. We further examined the polysaccharide production of VHb-expressing strain at different aeration levels in Erlenmeyer flasks. Again, in all cases, a significant enhancement of gellan production was observed, and the enhancement was more significant under oxygen-limiting conditions (up to 26·8%). VHb exhibited positive effect on cell growth and gellan yield of S. elodea, especially under hypoxic conditions. This is the first application of VHb as an effective metabolic engineering strategy in S. elodea to regulate cell growth and optimize gellan yield.

Sphingomonas paucibilis NK-2000 균주가 생산하는 젤란의 생산 농도 향상을 위한 포도당 첨가 및 교반속도와 통기량 변화 방법의 최적화

Sphingomnas paucibilis NK2000 균주를 사용한 젤란의 최적 교반속도 및 통기량은 각각 400 rpm 및 1.0 vvm이었다. 이 균주를 사용하여 젤란의 생산성을 향상시키기 위한 포도당의 최적 첨가시기는 배양을 시작한 36시간이었다. 젤란의 생산성을 향상시키기 위한 5가지 방법, 1) 포도당을 첨가하지 않는 방법, 2) 배양 36시간 후에 포도당을 첨가하지만 교반속도 및 통기량을 변화시키지 않는 방법, 3) 배양 36 시간 후에 포도당을 첨가하고 교반속도를 400 rpm에서 600 rpm으로 변화시키는 방법, 4) 배양 36 시간 후에 포도당을 첨가하고 교반속도를 400 rpm에서 600 rpm으로 증가시키고 통기량을 1.0 vvm에서 1.5 vvm으로 증가시키는 방법 및 5) 배양 36 시간 후에 포도당을 첨가하고 교반속도를 400 rpm에서 600 rpm으로 증가시키고 통기량을 1.0 vvm에서 2.0 vvm으로 증가시키는 방법 등을 실험한 결과, 젤란의 생산성은 각각 5.19, 5.74, 6.73, 7.93, 및 9.40 g/l이었으며, 변환률은 각각 26.0, 19.1, 22.4, 26.4, and 31.3%이었다. 최적의 방법으로 생산한 젤란의 생산농도 및 포도당 전환율은 포도당을 첨가하지 않은 방법으로 생산한 젤란의 생산농도 및 포도당 전환율에 비하여 각각 1.81 및 1.20 배 증가하였다. Optimal agitation speed and aeration rate for the production of gellan by Sphingomnas paucibilis NK2000 in a 7 l bioreactor were found to be 400 rpm and 1.0 vvm. The best time for glucose feeding into the medium for enhanced production of gellan by S. paucibilis NK2000 was 36 hr after cultivation. The concentrations of gellan produced by S. paucibilis NK2000 from 1) 20.0 g/l glucose without additional feeding, 2) 20.0 g/l glucose with feeding of 200.0 g/l glucose at 36 hr, in which the final concentration in the medium was 10.0 g/l, 3) 20 g/l glucose with feeding of 200.0 g/l glucose and a shift in an agitation speed from 400 to 600 rpm, 4) 20.0 g/l glucose with feeding of 200.0 g/l glucose at 36 hr and shifts in an agitation speed from 400 to 600 rpm and an aeration rate from 1.0 to 1.5 vvm, 5) and 20.0 g/l glucose with feeding of 200.0 g/l glucose at 36 hr and shifts in an agitation speed from 400 to 600 rpm and an aeration rate from 1.0 to 2.0 vvm, were 5.19, 5.74, 6.73, 7.93, and 9.40 g/l, respectively, and their conversion rates from glucose were 26.0, 19.1, 22.4, 26.4, and 31.3%, respectively. Compared to those developed using a normal process, production of gellan by S. paucibilis NK2000 from 20.0 g/l glucose was 1.81 times higher, and and its conversion rate was 1.20 times higher when the optimized process developed in this study was used.

Sphingomonas paucibilis NK-2000에 의한 젤란의 생산에 미치는 pH 안정제로서의 인산칼슐의 영향

Maximal productions of gellan by Sphingomnas paucibilis NK2000 from 20 g/l glucose and 10 g/l soybean pomace were 7.46 g/l in a flask and 7.35 g/l in a 7 l bioreactor, when the initial pH of media was 6.8. Maximal production of gellan in a 7 l bioreactor under pH control by sodium hydroxide was 8.42 g/l, whereas that under control by potassium phosphate was 8.50 g/l. The optimal concentration of potassium phosphate in a medium for production of gellan by S. paucibilis NK2000 was found to be 5.0 g/l. Maximal production of gellan in a medium containing 5.0 g/l potassium phosphate without pH control was 8.93 g/l in a 7 l bioreactor. In this study, a simple process without pH control was developed to enhance the production of gellan, with optimized concentration of potassium phosphate in the medium.

Occurrence, production, and applications of gellan: current state and perspectives

Bacterial exopolysaccharides (EPS) are products of biotechnology that are of high interest due to their rheological properties. This is the case of sphingans, a group of structurally related EPS secreted by members of the genus Sphingomonas. Among these, gellan is a multifunctional gelling agent produced in high yields by the non-pathogenic strain Sphingomonas elodea ATCC 31461. In its native form, gellan is a linear anionic EPS based on a tetrasaccharide repeat unit composed of two molecules of D: -glucose, one of L: -rhamnose and one of D: -glucuronic acid. The native gellan is partially esterified with acyl substituents (1 mol of glycerate and 0.5 mol of acetate) per repeat unit. Gellan has unique characteristics and has many applications, particularly in the food, pharmaceutical, and biomedical fields. This review summarizes current knowledge on the structure and properties of gellan and provides details about the biosynthesis of this exopolysaccharide. In addition, a highlight of the importance of gellan in industrial and medicinal applications is given.

Production and rheological characteristics of the microbial polysaccharide gellan

B. MANNA, A. GAMBHIR AND P. GHOSH. 1996. Sphingomonas paucimobilis was used for the synthesis of the microbial polysaccharide gellan. In a 60 h fermentation, polysaccharide yield and productivity obtained were 0.45 g gellan per g glucose consumed and 0.21 g I−1 h−1 respectively. The broth showed pseudoplastic behaviour with yield stress. The requirement of the solvent propanol to precipitate gellan from the broth depended on the volume of the broth rather than on gellan concentration. The addition of salt to the broth reduced the propanol requirement. Attempts to separate cells from the highly viscous broth by microfiltration were not successful. Ultrafiltration reduced the propanol requirement but appreciable membrane fouling and loss of gellan was observed.

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.

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.

The effect of agitation and aeration on the synthesis and molecular weight of gellan in batch cultures of Sphingomonas paucimobilis

The effects of agitation and aeration upon synthesis and molecular weight of the biopolymer gellan were systematically investigated in batch fermenter cultures of the bacterium, Sphingomonas paucimobilis. High aeration rates and vigorous agitation enhanced growth of S. paucimobilis. Although gellan formation occurred mainly in parallel with cell growth, the increase in cells able to synthesise gellan did not always lead to high gellan production. For example, at very high agitation rates (1000 rpm) growth was stimulated at the expense of biopolymer synthesis. Maximal gellan concentration was obtained at 500 rpm agitation and either 1 or 2 vvm aeration (12.3 and 12.4 g/l gellan, respectively). An increase in aeration (from 1 to 2 vvm) enhanced gellan synthesis only at low agitation rates (250 rpm). However, high aeration or dissolved oxygen was not necessary for high gellan synthesis, in fact oxygen limitation always preceded the phase of maximum gellan production and probably enhanced polysaccharide biosynthesis. Some gellan was formed even after glucose exhaustion. This was attributed to the intracellular accumulation of polyhydroxyalkanoates, (such as polyxydroxybutyrate) which were found in S. paucimobilis cells indicating the existence of a carbon storage system, which may contribute to gellan biosynthesis under glucose-limiting conditions. The autolysis of the culture, which occurred at the late stages of the process, seemed to be triggered mainly by limitations in mass (nutrient) transfer, due to the highly viscous process fluid that gradually develops. Rheological measurements generally gave a very good near real time estimate of maximum biopolymer concentration offering the possibility of improved process control relative to time consuming gravimetric assay methods. While mechanical depolymerisation of gellan did not occur, high aeration rates (2 vvm) led to production of gellan of low molecular weight (at either 250 or 500 rpm). This effect of aeration rate upon gellan molecular weight is reported here for the first time, and is important for the properties and applications of gellan. Mechanisms which may have led to this are discussed, but control of molecular weight of the biopolymers is clearly an area needing further research.