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 (Accession No. GI:724472387) and (GI:724472388) by 16S rRNA gene sequencing. To maximize gellan production by and, 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 than 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 52.6 g/l, respectively) using WG (80 g/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, and demonstrate great potential for use in the commercial production of gellan and in the bioremediation of WG.
Glycerol, a non-biodegradable by-product during biodiesel production is a major concern to the emerging biodiesel industry. Many microbes in natural environments have the ability to utilize glycerol as a sole carbon and energy source. The focus of this study was to screen for microorganisms from soil, capable of glycerol utilization and its conversion to value added products such as ethanol and 1,3-propanediol (1,3-PDO). Twelve bacterial isolates were screened for glycerol utilization ability in shake flask fermentations using M9 media supplemented with analytical grade glycerol (30 g/L) at various pH values (6, 7 and 8) and temperatures (30°C, 35°C and 40°C). Among these, six bacterial isolates (SM1, SM3, SM4, SM5, SM7 and SM8) with high glycerol degradation efficiency (>80%) were selected for further analysis. Highest level of 1,3-PDO production (15 g/L) was observed with isolate SM7 at pH 7 and 30°C, while superior ethanol production (14 g/L) was achieved by isolate SM9 at pH 8 and 35°C, at a glycerol concentration of 30 g/L. The selected strains were further evaluated for their bioconversion efficiency at elevated glycerol concentrations (50-110 g/L). Maximum 1,3-PDO production (46 g/L and 35 g/L) was achieved at a glycerol concentration of 70 g/L by isolates SM4 and SM7 respectively, with high glycerol degradation efficiency (>90). Three isolates (SM4, SM5 and SM7) also showed greater glycerol tolerance (up to 110 g/L). The isolates SM4 and SM7 were identified as Klebsiella pneumoniae and SM5 as Enterobacter aerogenes by 16S rDNA analysis. These novel isolates with greater glycerol tolerance could be used for the biodegradation of glycerol waste generated from the biodiesel industry into value-added commercial products.
The continual growth of the global biodiesel industry has resulted in a proportional increase in crude glycerol production. The by-production of glycerol waste during the manufacture of biodiesel has, with recent research, proven to hold use as a feedstock for the production of several commodity chemicals. The conversion of glycerol may be carried out by both chemical as well as biological means. The biological conversion of glycerol surpasses chemical conversion with respect to higher yield and selectivity, normal reaction conditions and the use of cheaper biological catalysts. Many microorganisms are known to convert glycerol to different value added products. This study involved the isolation of bacteria from soil and crude glycerol from a local biodiesel plant. Isolates were then used to convert crude glycerol supplemented with salts and a nitrogen source into commercially viable products. Isolates which successfully degraded glycerol were then identified via 16S PCR. A strain of Klebsiella pneumoniae, which is a known producer of 1,3-propanediol (1,3-PDO), was isolated from soil and two strains of Sphingomonas sp., which is a known gellan producer, was isolated from biodiesel waste. Gellan is an exopolysaccharide used in the food, cosmetic and pharmaceutical industries sold commercially as a product known as Gelrite or Gelzan while 1,3-PDO is an important component of fuels and polyesters (used widely in the petroleum industry) and is currently chemically produced. Using crude glycerol for producing 1,3-PDO is a good solution from an economic as well as ecological point of view. K. pneumoniae, Sphingomonas psueudosanguinis and Sphingomonas yabuuchiae were subjected to a series of shake flask fermentations in order to determine optimal growth conditions. This microoganism was able to successfully produce significant amounts of 1,3-PDO and lactic acid using crude glycerol (80 g/l), without pre-treatment (37 and 6.8 g/l respectively). S psueudosanguinis and S. yabuuchiae were both able to produce two of the highest amounts of gellan gum than that reported by other studies using crude glycerol (80 g/l) as a sole carbon source in a minimal medium (50.9 and 52.6 g/l respectively).
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