INTRODUCTION LAS was the first surfactant introduced to solve an enviromental problem. LAS is one of synthetic surfactants that is widely used. Synthe

Transcription

1 BIODEGRADATION OF LINEAR ALKYLBENZENE SULFONATE BY BACTERIAL CONSORTIUM Praswasti PDK Wulan, Misri Gozan, Anondho Wijanarko, Dianursanti,, Marisa Tejo, Mahmud Sudibandriyo

2 INTRODUCTION LAS was the first surfactant introduced to solve an enviromental problem. LAS is one of synthetic surfactants that is widely used. Synthetic type surfactants are often used because they perform better and more economical compared to natural detergents Although LAS is biodegradable, its introduction to the environment in big amount harms the water bodies. People assume that detergents performing bulk of foam represent the quality of detergents. In nature pseudomonas live contiguous with bacillus Using Bioremediation to degradate polutants in environment. The advantages are: no side effect, more economic compare with other methods Pusat Penelitian dan Pengembangan Teknologi Minyak dan Gas Bumi (LEMIGAS) has found mixtures of microorganism (bacterial consortium) in sewage plant that degrade oil pollutants

3 Chemical structure of LAS : have linear chain Critical Micelle Concentration (CMC) : Limiting concentration of surfactant for beginning a micelle formation which consist of several hundred collected surfactant monomers (Hart et al, 2003). micelle from LAS in water (Hart, et. al, 2003) the surfactant solution consists monomer in equilibrium condition and micelle form that contribute to stop the decreasing of solution surface tension (Rosen, 1989) CMC visualized a minimum surface tension (Rosen, 1989)

4 Aerob degradation Metabolism of Linear AlkylBenzene Sulfonate (LAS) (Schkeheck, 2003) A. Aerob Biodegradation: LAS will be change to SPC (sulphophenyl Carboxilates) = intermediate product. B. Aromatic chain will be total conversion of water, carbondioxide and sulphate

5 Pseudomonas aeroginosa Bacillus cereus (Scimat, 2005) Bacillus subtilis

6 Pure Cultivation Bacterial consortium which was collected from LEMIGAS had been cultivated purely with nutrient broth medium METHODOLOGY Enrichment of bacterial consortium in Lockhead and Chase (LC) medium To o adaptate the bacterial consortium toward LAS. Taking data and analysis Total Plate Count (TPC) which was used for counting bacteria inoculum. Measuring LAS concentration with Standard Test Method for Methylene Blue Active Substances (ASTM D 2330). Measuring test for Chemical Oxygen Demand (COD).: : to represent degradation LAS in system because LAS was the only organic chemistry in this system and to detect suitability of water. Degradating implementation of bacteria consortium towards LAS This step took time in 264 hours (12 days) in 250 ml erlenmeyer, 22 rpm with several variation concentration of LAS: 100, 400, 700, 1000, 1500 ppm. These concentrations were chosen in consideration of using amount of LAS in detergent.

7 Extraction at Methylene Blue Active Substances standard Methode : to measure LAS concentration LAS SOLUTION (LAS Solution + methylene blue) Methylene blue + LAS (anionic surfactant) blue color Extraction result with chloroform: 2 phase in LAS solution + methylene blue Callibration curve : absorbansi vs LAS concentration (ppm) has to be made before Turbidity was observed with Spektrophotometer with 650 nm wavelength

8 EXPERIMENT RESULT Cultivation : to get growth of bacteria data + growth rate - growth rate = - 0,032/h Optimum Growth of Bacteria Consortium in Nutient Both Medium = hour-1 CFU/mL 1.E+09 1.E+08 1.E+07 1.E+06 1.E+05 1.E+04 1.E+03 1.E+02 1.E+01 1.E hours It means growth rate of bacteria vanishes Biodegradation step can be running until 264 hour (12 days)

9 Growth of Bacteria Consortium in Enrichment Step : The first enrichment The first adaptation bacterial consortium toward LAS to o adaptate the bacterial consortium toward LAS and using LAS as a carbon sources for its metabolism Start to death phase Leads to decrease amount of bacteria because their adapting phase in new environment which contains of LAS. Sudden contact of bacteria consortium with relatively high LAS could disturb their living to maintain carbon source for bacteria growth. added 100 ppm periodically: to ensure that the consortium could acclimatize with higher concentration of LAS. to definite there was no lack of carbon source. This addition increase amount of consortium. It means that the consortium could acclimatize with LAS Optimum Growth of Bacteria Consortium = hour-1 1 at 360 h

10 Amount of Bacteria Consortium in Various LAS Concentration Amount of Bacteria Consortium 3.00E E E E E E E+00 Standard of comparison system t (hours) Bacteria do not need long time period compare to the other concentrations for adapt well with new environment. It can reach peak condition and stationer condition. After reaching maximum amount of population, there was lack of nutrition, and then the consortium started to reach death phase. 0 ppm 100 ppm 400 ppm 700 ppm 1000 ppm 1500 ppm 1. Bacteria consortium can live in LAS environment. The use of LC medium which contains adequate nutrition made the consortium could live and grow. 2. Bacteria decrease at the first time biodegradation process because they have to adapt well with new environment (enter to growth phase). Weak bacteria will be dead. 3. Bacteria that can adapt well will be growth until reach its peak. In this condition LAS biodegradation will increase until maximum condition. 4. Growing phase pattern of bacteria consortium relatively more unclear. After they had reached maximum population, they shortly entered death phase. The stationary phase relatively unclear at higher LAS concentration.

11 Optimum Growth Rate of Bacteria in Variated LAS Concentration LAS Concentration (ppm) Optimum Growth Rate (per hours) Several factors that may affect grow of bacteria in this research are : Agreement of bacterial consortium with LC medium. Growing time period which did not same for each bacteria in this consortium. LAS could be toxic to some bacteria. Competition with each bacteria.

12 Relation of LAS Concentration with Surface Tension : CMC LAS = 513,98 ppm. Below CMC : higher LAS concentration, lower surface tension will be happened. Surface tension has already reached minimum number and start to be steady at LAS concentration above CMC. 0 ppm >100 ppm >400 ppm >700 ppm, LAS 700 ppm = 1000 ppm = 1500 ppm. Addition of LAS with larger concentration influenced bacteria s living environment. The first adaptation of bacteria with new environment: They had to do adaptation hardly. Their ability to do metabolism by explored LAS as carbon source and agreement with their living medium which might toxic to them Bacteria adapt well with environment that they can reproduce and utilize LAS as a carbon source

13 COD in LAS Biodegradation Process COD (ppm) t (hours) COD symbolizes oxygen which is needed for degradating organic chemical in water. This figure show decreasing pattern of COD in all system with variated LAS concentration. This indicated that decreasing concentration of LAS or LAS biodegradation process by bacteria consortium was trully happened.

14 Result of Degradation COD Research in Various LAS Concentration : LAS Concentration Total Degradation COD (%) 100 ppm ppm ppm ppm ppm COD that measured : amount oxygen is needed by contaminant in system. Can not see contaminant in LAS form or intermediate product biodegradation by bacteria. COD = amount of oxigen needed that degradation of organic substances in water. Lower LAS concentration higher COD decreasing COD value was not greatly reduced. It means that biodegradation had happened but there still a lot of carbon source available whether in form of LAS or intermediate form. All this organic carbons was counted as COD value. Using this Bacteria consortium have advantages : when there were lot of bacteria died because of toxic material, there is still another bacteria could hold out and do the degradation.

15 CONCLUSION The consortium of bacteria (Pseudomonas( aeroginosa, Bacillus subtilis, Bacillus aglomerans, Bacillus cereus, Bacillus alvae) ) could survive until 1500 ppm of LAS. While at higher LAS concentration the stationery phase was relatively unclear. LAS concentration above CMC,, surface tension has already reached minimum number and start to be steady. Below CMC, higher LAS concentration lower surface tension will be happened : 0 ppm >100 ppm >400 ppm >700 ppm, LAS 700 ppm = 1000 ppm = 1500 ppm. Bacteria consortium that was used in this research could degradate LAS but less efficient to reducing COD value in those systems.