Bio-Assisted Process for the treatment and regeneration of spent caustic

Abstract

The present invention relates to a bio-assisted method for treatment of spent caustic by treating with haloalkaliphilic consortium of bacteria capable of reducing or transforming sulphides, thiols, mercaptants and other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

Claims

1. A bio-assisted method of treating spent caustic using haloalkaliphilic bacterial consortium said method comprises the steps of: (a) mixing the spent caustic with haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389) and Lysinibacillus sp. (MTCC 5666) in a reactor; (b) maintaining the percentage of saturated oxygen in the reactor initially in the range of 0-20% or 60-100% saturation for 48 hrs followed by saturated oxygen in the range of 60-100% or 0-20% for 24-48 hrs respectively; (c) carrying out the reaction of steps (a) and (b) for about 3-5 days; and (d) obtaining caustic with reduced concentration of sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives.

2. The method as claimed in claim 1, wherein in step (a) is performed at the pH in the range of 8 to 13.

3. The method as claimed in claim 1, wherein in step (b) the saturated oxygen in the reactor is initially maintained in the range of 0-20% followed by saturated oxygen maintained in the range of 60-100%.

4. The method as claimed in claim 1, wherein in step (b) the saturated oxygen in the reactor is initially maintained in the range of 60-100% followed by saturated oxygen maintained in the range of 0-20%.

5. The method as claimed in claim 1, wherein the haloalkaliphilic bacterial consortium is present in a concentration of at least 10.sup.2 CFU/ml.

7. The method as claimed in claim 1, wherein said method is done either in same reactor by altering the air saturation after certain time or in two series reactor where effluent of one reactor work as influent to another reactor and both reactors has different air saturation level.

8. The method as claimed in claim 1, wherein haloalkaliphilic bacterial consortium is used in immobilized or free form.

9. The method as claimed in claim 1, wherein said method is performed in batch mode as well as continuous mode using continuously stirrer reactor, up-flow reactor and any such suitable continuous mode reactor.

10. The method as claimed in claim 1, wherein the haloalkaliphilic bacterial consortium are immobilized when the reaction is carried out in continuous mode.

11. The method as claimed in claim 1, wherein the reaction is carried out in a single reactor or two reactors.

12. A haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

13. The haloalkaliphilic consortium of bacteria as claimed in claim 8, wherein the said haloalkaliphilic consortium is capable of treating spent caustic.

14. Use of haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

15. Use of haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) in a method of for reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

Description

DETAILED DESCRIPTION

[0032] While the invention is susceptible to various modifications and/or alternative processes and/or compositions, specific embodiment thereof has been shown by way of example in the drawings and tables and will be described in detail below. It should be understood, however that it is not intended to limit the invention to the particular processes and/or compositions disclosed, but on the contrary, the invention is to cover all modifications, equivalents, and alternative falling within the spirit and the scope of the invention as defined by the appended claims.

[0033] The tables and protocols have been represented where appropriate by conventional representations, showing only those specific details that are pertinent to understanding the embodiments of the present invention so as not to obscure the disclosure with details that will be readily apparent to those of ordinary skill in the art having benefit of the description herein.

[0034] The following description is of exemplary embodiments only and is not intended to limit the scope, applicability or configuration of the invention in any way. Rather, the following description provides a convenient illustration for implementing exemplary embodiments of the invention. Various changes to the described embodiments may be made in the function and arrangement of the elements described without departing from the scope of the invention.

Definitions

[0035] The term “Haloalkaliphilic bacteria” as used in the context of the present invention means bacteria is able to grow under high alkaline conditions which comprises of high salt as well as high pH which is above pH 8. More specifically such “Haloalkaliphilic bacteria” grow at pH in the range of 8-13. The “Haloalkaliphilic bacteria” have a special mechanisms to survive and grow under salinity and high alkaline pH wherein these bacteria can survive at pH in the range of 8 to 13 and salt concentration of 2-8%.

[0036] The term “Haloalkaliphilic consortium” or “Haloalkaliphilic consortium of bacteria or “Haloalkaliphilic bacterial consortium”, as used in context of the present invention refers to consortium or mixture of bacteria selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of treating or breaking down contaminants from the spent caustic. The “Haloalkaliphilic consortium” or “Haloalkaliphilic consortium of bacteria or “Haloalkaliphilic bacterial consortium” or “microbial consortium” in the context of present invention could also means a consortium or mixture of any two or more bacteria selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of treating or breaking down contaminants from the spent caustic.

[0037] The term “Contaminants” as used in context of present invention refers to all the impurities present in the spent caustic not limited to but including sulphides, thiols, mercaptants and other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives.

[0038] The term “Ambient temperature” and “pressure” as used in context of the present invention relates to the temperature and pressure ranging from 20-40° C. and 1-2 bar, respectively.

[0039] The term “Hydraulic Retention Time (HRT)” as used in context of the present invention relates to average length of time that a spent caustic remains in the reactor.

[0040] The term “Industrial Application” as used in context of the present invention refers to use of treated spent caustic in industrial processes for removal of contaminants from gaseous and hydrocarbon streams in oil refinery processes or may be used for the pH maintenance of the waste water treatment plans or biological processes.

[0041] The term “Oxygen saturation” or “percentage of oxygen saturation” as used in context of the present invention refers to relative measure of the amount of oxygen that is dissolved or carried in a given medium. In context of present invention the percentage of oxygen saturation can be extensively varied at the start or initial stage of the treatment as well at the later stage of the treatment, as discussed below: [0042] (i) At the start of the treatment the oxygen saturation can be maintained as low as in the range of 0-20% followed by higher oxygen saturation in the range of 60-100%; or [0043] (ii) At the start of the treatment the oxygen saturation can maintained as high as in the range of 60-100% followed by lowering of the oxygen saturation in the range of 0-20%.

[0044] As used herein, the terms “Oxygen saturation” or “percentage of oxygen saturation” as used in the context of the present invention have been used interchangeably and are meant to have the same definition and meaning as herein described.

[0045] The main embodiment of the present invention provides a bio-assisted method of treating spent caustic using haloalkaliphilic bacterial consortium said method comprises the steps of: [0046] (a) mixing the spent caustic with haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389) and Lysinibacillus sp. (MTCC 5666) in a reactor; [0047] (b) maintaining the percentage of saturated oxygen in the reactor initially in the range of 0-20% or 60-100% saturation for 48 hrs followed by saturated oxygen in the range of 60-100% or 0-20% for 24-48 hrs respectively; [0048] (c) carrying out the reaction of steps (i) and (ii) for about 3-5 days; and [0049] (d) obtaining caustic with reduced concentration of sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives.

[0050] Another embodiment of the present invention provides a bio-assisted method of treating spent caustic using haloalkaliphilic bacterial consortium said method comprises the steps of: [0051] (a) mixing the spent caustic with haloalkaliphilic consortium of any two or more bacteria selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389) and Lysinibacillus sp. (MTCC 5666). and nutrient system in a reactor; [0052] (b) maintaining the percentage of saturated oxygen in the reactor initially in the range of 0-20% or 60-100% saturation for 48 hrs followed by saturated oxygen in the range of 60-100% or 0-20% for 24-48 hrs respectively; [0053] (c) carrying out the reaction of steps (i) and (ii) for about 3-5 days; and [0054] (d) obtaining caustic with reduced concentration of sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives.

[0055] Another embodiment of the present invention provides a method as herein described wherein the said haloalkaliphilic bacterial consortium in step (a) comprises any three or more bacteria selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389) and Lysinibacillus sp. (MTCC 5666).

[0056] Another embodiment of the present invention provides haloalkaliphic bacterial consortium used for the treatment of spent caustic to remove sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, napthenic acids and their derivatives from the spent caustic, wherein the contaminants in spent caustics under conditions of ambient temperature and pressure.

[0057] In another aspect the present invention provides a method of treating contaminants of spent caustic with haloalkaliphic bacterial consortium which are having pH in the range of 8-13 and wherein haloalkaliphic bacterial consortium can tolerate a temperature in the range of 5−60° C.

[0058] Another embodiment of the present invention provides a method as herein described wherein the method in step (a) is performed at pH in the range of 8 to 13.

[0059] Another embodiment of the present invention provides a method as herein described wherein the method in step (a) is performed above pH 8.

[0060] Yet another embodiment of the present invention provides a method as herein described wherein the haloalkaliphilic microbial consortium is present in a concentration of 10.sup.2 CFU/ml.

[0061] Another embodiment of the present invention provides a method as herein described wherein in step (b) the saturated oxygen in the reactor is initially maintained in the range of 0-20% followed by saturated oxygen maintained in the range of 60-100%.

[0062] Another embodiment of the present invention provides a method as herein described wherein in step (b) the saturated oxygen in the reactor is initially maintained in the range of 60-100% followed by saturated oxygen is in the range of 0-20%.

[0063] Another embodiment of the present invention provides a method as herein described wherein then method is carried out in two reactors then saturated oxygen in the first reactor is maintained at 0-20% or 60-100% followed by saturated oxygen in the second reactor is maintained at 60-100% or 0-20% respectively.

[0064] Another embodiment of the present invention provides a method as herein described wherein then method is carried out in single reactor, wherein the saturated oxygen is initially maintained at 0-20% or 60-100% followed by saturated oxygen maintained at 60-100% or 0-20% respectively.

[0065] Another embodiment of the present invention provides a method as herein described wherein the haloalkaliphilic bacterial consortium is present in a concentration of at least 10.sup.2 CFU/ml.

[0066] Another embodiment of the present invention provides a method as herein described wherein the haloalkaliphilic bacterial consortium is used in immobilized or free form.

[0067] Another embodiment of the present invention provides a method as herein described wherein said method is done either in same reactor by altering the air saturation after certain time or in two series reactor where effluent of one reactor work as influent to another reactor and both reactors has different air saturation level.

[0068] Yet another embodiment of the present invention provides method as herein described wherein the method is performed in batch mode as well as continuous mode using continuously stirrer reactor, up-flow reactor and any such suitable continuous mode reactor.

[0069] Yet another embodiment of the present invention provides a method as herein described wherein the haloalkaliphilic bacterial consortium are immobilized when the reaction is carried out in continuous mode.

[0070] Yet another embodiment of the present invention provides a method as herein described wherein the reaction is carried out in a single reactor or two reactors.

[0071] Yet another embodiment of the present invention provides haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

[0072] Yet another embodiment of the present invention provides two or more haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

[0073] Yet another embodiment of the present invention provides haloalkaliphilic consortium of bacteria wherein the said haloalkaliphilic consortium is capable of treating spent caustic.

[0074] Yet another embodiment of the present invention provides use of haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

[0075] Yet another embodiment of the present invention provides use of two or more haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) capable of reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

[0076] Yet another embodiment of the present invention provides use of haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) in a method for reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

[0077] Yet another embodiment of the present invention provides use of two or more haloalkaliphilic bacterial consortium selected from Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389), Lysinibacillus sp. (MTCC 5666) in a method for reducing or transforming sulphides, thiols, other sulphur containing compounds, phenols, hydrocarbons, naphthenic acids and their derivatives in spent caustic.

[0078] In another aspect of the method of present invention the treatment of spent caustic with haloalkaliphilic bacterial consortium is carried out at percentage of saturated oxygen, wherein the percentage of oxygen saturation can be extensively varied at the start or initial stage of the treatment as well at the later stage of the treatment, as discussed below. The said aspect of the invention has been demonstrated in Examples 4 and 6. [0079] (i) At the start of the treatment the oxygen saturation can be maintained as low as in the range of 0-20% followed by higher oxygen saturation in the range of 60-100%; or [0080] (ii) At the start of the treatment the oxygen saturation can maintained as high as in the range of 60-100% followed by lowering of the oxygen saturation in the range of 0-20%.

[0081] Another embodiment of the present invention provides a method as herein described wherein the percentage of saturated oxygen in the reactor is initially maintained as low as in the range of 0-20% followed by a higher percentage of saturated oxygen in the range of 60-100% (Example 4).

[0082] Yet another embodiment of the present invention provides a method as herein described wherein the percentage of saturated oxygen in the reactor is maintained for 5-48 hrs at 0-20% saturation followed by 60-100% saturation for 10-48 hrs.

[0083] Yet another embodiment of the present invention provides a method as herein described wherein the percentage of saturated oxygen in the reactor is maintained for 30 hrs at 10% saturation followed 100% saturation for 30 hrs.

[0084] Another embodiment of the present invention provides a method as herein described wherein the percentage of saturated oxygen in the reactor is maintained for 5-48 hrs at 60-100% saturation followed by 0-20% saturation for 10-48 hrs.

[0085] Yet another embodiment of the present invention provides a method as herein described wherein the percentage of saturated oxygen is the reactor is maintained for a week at 0-20% saturation followed by 60-100% saturation.

[0086] In another aspect the present invention provides a method wherein the treatment of spent caustic can be carried out in a single reactor or two reactors. When the reaction is carried out in two reactors then product of first reactor is transferred to the second reactor. In case of two reactor both reactors are maintained at different oxygen saturation concentrations i.e. in the first reactor the oxygen saturation is in the range of 0-20% saturation and in the second reactor the oxygen saturation is in the range of 60-100% saturation.

[0087] Further another aspect of the present invention provides that when reaction is carried in two reactors in series then the reaction is run in continuous mode, whereas when the reaction is carried out in a single reactor the reaction is run in batch mode.

[0088] Another aspect of the present invention provides a reactor system in which the exhaust gas that is stripped from the reactor suspension, containing volatile compounds like sulfur dioxide, hydrogen sulfide, hydrocarbons and their and metabolites are continuous recycled to the reactor after condensing them in a condenser maintained at temperature of 5° C.

[0089] In another aspect the present invention provides a bio-assisted method wherein the disposal and regeneration of spent caustic is performed at an ambient temperature and pressure.

[0090] Yet another embodiment of the present invention after treatment of the spent caustic the biomass can be recovered by suitable technique like centrifugation, filtration and the recovered biomass can be re-used for treatment of fresh spent caustic.

[0091] Another aspect of the present invention provides a method as herein described wherein the mixture of spent caustic and haloalkaliphilic consortium treated in batch mode as well as continuous mode using a stirrer reactor, up-flow reactor and such suitable reactor.

[0092] In another embodiment the present invention provides a method in which after treatment of spent caustic using the haloalkaliphilic consortium as herein described in present invention the recovered caustic free of contaminants can be used for various purposes like maintenance of pH of ETPs, removal of contaminants from gaseous and hydrocarbon streams.

[0093] Another aspect of the present invention provides a method of treatment of spent caustic as herein described wherein the method uses a nutrient system consisting of K.sub.2HPO.sub.4 (4-10 g/l), KH.sub.2PO.sub.4 (4-10 g/l), MgCl.sub.2 (0.2-2 g/l), 0.5-2 ml trace elements, sodium carbonate (2-20 g/l), yeast extract (4-10 g/l), ammonium nitrate (5-8 g/l), citrate (10-20 g/l), sorbitol ester (5-25 ppm), Oleic acid (100-1000 ppm), pantothenic acid (20-500 ppm), thiamine (25-300 ppm). The trace element solution (gram per liter) comprises nitrilotriacetic acid (1.0), FeSO.sub.4.7H.sub.2O (0.01), MnCl.sub.2.4H.sub.2O (0.005), CoCl.sub.2.6H.sub.2O (0.09), CaCl.sub.2.2H.sub.2O (0.9), ZnCl.sub.2 (0.55), CuCl.sub.2.H.sub.2O (0.03), H.sub.3BO.sub.3 (0.02), Na.sub.2MoO.sub.4 (0.02).

[0094] The invention will now be explained with the help of following examples. However, the scope of the invention should not be limited to these examples as the person skilled in the art can easily vary the proportion of the ingredients and combinations.

Example 1: Isolation of the Microbes

[0095] The Microbes were Isolated by Enrichment:

[0096] Microbes were isolated through enrichment culture techniques and directed forced evolution. Briefly, to isolate the microbes with desired traits activated sludge from aeration tank of refinery (Indian Oil, Panipat Refinery Panipat, Haryana, India) effluent treatment plant was added to spent caustic obtained from refinery after amending with growth supplements. The growth supplements contained (g/l) Na.sub.2CO.sub.3 (5), NaHCO.sub.3 (2.5) KH.sub.2PO.sub.4 (4), K.sub.2HPO.sub.4 (1), MgSO.sub.4 (1.0), (NH.sub.4).sub.2SO.sub.4 (0.50), KNO.sub.3 (2.0), ZnSO.sub.4 (0.5), yeast extract (4), and trace element (2 ml solution). The trace element solution (gram per liter) comprises Nitrilotriacetic acid (1.0), FeSO.sub.4.7H.sub.2O (0.01), MnCl.sub.2.4H.sub.2O (0.005), CoCl.sub.2.6H.sub.2O (0.02), CaCl.sub.2.2H.sub.2O (0.5), ZnCl.sub.2 (0.15), CuCl.sub.2.H.sub.2O (0.03), H.sub.3BO.sub.3 (0.02), Na.sub.2MoO.sub.4 (0.02), Na.sub.2SeO.sub.3 (0.02), NiSO.sub.4 (0.03), SnCl.sub.2 (0.03). The contaminants present spent caustic served as only source of carbon and energy. The culture system incubated aerobically at temperature ranging from 30-50° C. At weekly interval it was transfer to fresh media also using refinery spent caustic as sole carbon and energy source. This was repeated for 7 cycles and subsequently microbes were isolated on agar plate having pH 14 and salinity 4%. The bacterial colonies appeared on the agar plate were further purified. Subsequently, individual bacteria are grown on refinery spent caustic and its growth is evaluated at timed interval by measuring colony forming units (CFU) on agar plate. Bacteria are selected for further studies based on their ability to grow faster and degrade at least one of the contaminants at high pH and salinity. The bacteria which were found suitable for the present invention comprises of Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024). These bacteria were used in combination with hydrocarbon degrading Bacillus substilis (MTCC 5386), Pseudomonas aeruginosa (MTCC 5389) and Lysinibacillus sp. (MTCC 5666).

[0097] The foresaid microbes have been deposited with the Microbial Type Culture Collection (MTCC), Chandigarh, India as required under the Budapest Treaty and have been assigned there respective Accession numbers as recited above.

Example 2: Development of the Consortia

[0098] Based on growth ability, a combination of bacteria including Pseudomonas stutzeri (MTCC 25027), Arthobacter sp. (MTCC 25028), Bacillus subtilis (MTCC 25026), Achromobacter xylooxidan (MTCC 25024), Lysinibacillus sp. (MTCC 5666) are developed. The consortium showed synergistic effect in terms of degradation of contaminants present in spent caustic as represented in Table 1.

TABLE-US-00001 TABLE 1 The effect of Haloalkaliphilic bacterial consortium % hydrocarbon % phenol degradation degradation S. No. Bacteria/mixture in 50 hr in 50 hr 1 Pseudomonas stutzeri 63.3 23.4 (MTCC 25027), 2 Arthobacter sp. 42.2 52.8 (MTCC 25028), 3 Bacillus subtilis 71.1 45.1 (MTCC 25026), 4 Achromobacter xylooxidan 56.6 23.8 (MTCC 25024) 5 Lysinibacillus sp. 51.2 43.6 (MTCC 5666) 6 Consortium/Mixture of above 97.9 99.2 bacteria (as mentioned in S. Nos. 1-6 above) in equal proportion

[0099] The Haloalkaliphilic bacterial consortium was evaluated in shake flask at 35° C. and 120 rpm actual spent caustic obtained from refinery. The control without inoculation is used as negative control and is run under similar conditions. At the time interval samples are taken and analyzed for CFU/ml on agar plate as well for concentration of contaminant by suitable analytical techniques. The Haloalkaliphilic bacterial consortium also can be used in immbolized form or in free form. The Haloalkaliphilic bacterial consortium can be immobilized varieties of membranes such as synthetic plastics, surface-modified carbon nanotubes, poly (tetrafluoroethylene) (PTFE) fibrils, zeolite, clay, anthracite, porous glass, activated charcoal, ceramics, acrylamide, polyurethane, polyvinyl, resins and natural polymer etc. The advantage of using Haloalkaliphilic bacterial consortium in an immobilized form provides enhanced microbial cell stability, allows continuous process operation and prevents the requirement to separate the biomass-liquid separation requirement. The immobilization can be done as per the method known in prior art.

Example 3: Treatment in Bioreactor

[0100] The spent caustic is fed in a CSTR with air bubbling system reactor and to the reactor nutrient system containing K.sub.2HPO.sub.4 (4 g/l), KH.sub.2PO.sub.4 (4 g/l), MgCl.sub.2 (0.2 g/l), 0.5 g/l of trace elements, sodium carbonate (2 g/l), yeast extract (5 g/l), sodium nitrate (4 g/l), citrate (5-10 g/l), sorbitol ester (5 ppm), Oleic acid (100 ppm), pantothenic acid (20 ppm), thiamine (25 ppm) was added. The reactor is inoculated with haloalkaliphilic microbial consortium to the cell concentration of 10.sup.2 CFU/ml. The spent caustic was treated as received from an oil refinery and reactor was run at ambient temperature. The % of oxygen saturation of the reactor was maintained initially for 30 hr at 10% saturation followed by saturation at 100% level. The stirring of the reactor was adjusted at 50 rpm for initial 30 hrs followed by 800 rpm for next 30 hrs. To prevent the release of volatile compounds from the system, gas phases are continuously recycled. The recycled gas is first passed to a condenser (maintained at 5° C.) to recover the volatile compounds and metabolites. A control without bacteria was also run under similar conditions. The treatment was done without sterilizing the spent caustic.

[0101] At the time interval samples are taken and analyzed for CFU/ml on agar plate as well for concentration of contaminant by suitable analytical techniques as represented in Table 2.

TABLE-US-00002 TABLE 2 Haloalkaliphilic bacterial consortium count during contaminant removal process in refinery spent caustic Time Control without Inoculated with microbial (in hrs) inoculation consortia 0 Nil 7.20E+02 10 Nil 8.90E+07 20 Nil 7.80E+08 30 Nil 3.20E+11 40 Nil 3.80E+12 50 Nil 7.60E+14 60 Nil 8.80E+14

[0102] Table-2 shows the time dependent growth of the Haloalkaliphilic bacterial consortium indicating that they utilize contaminates present in the spent caustic as carbon and energy source. In the control almost no growth of bacteria was found up which showed that indigenous Haloalkaliphilic bacterial consortium has no role in the contaminant removal which is further corroborated by decrease in the concentration of the contaminates removal in the system where microbial consortia was added (Table-3).

[0103] The treated spent caustic has more than 98% reduction in total sulfur, sulphides, mercaptans, hydrocarbon, phenol and other contaminants in comparison to abiotic control without the Haloalkaliphilic bacterial consortium (as represented in Table 3). The pH of the reactor with Haloalkaliphilic bacterial consortium was 13 in comparison to 14 in abiotic control after 60 hrs. The treated spent caustic is centrifuged at 8000 rpm for 10 min and biomass s removed. Such obtained cell and contaminant free caustic can be reused. The biomass obtained after centrifugation is used for treatment of another batch of the spent caustic.

TABLE-US-00003 TABLE 3 The contaminant removal in refinery spent caustic % degradation Thiol Merceptans Phenol sulphides Control Control Control Control without without without without microbial With microbial microbial With microbial microbial With microbial microbial With microbial Time (hrs.) consortia consortia consortia consortia consortia consortia consortia consortia 10 0.1 33.2 0.1 28.3 0 43.4 1 32.4 20 0.2 75.4 0.2 70.4 0 63.6 2 43.8 30 0.2 92.4 0.3 90.7 0.1 93.4 3.1 67.5 40 0.6 97.7 0.7 87.4 0.3 95.8 3.3 85.4 50 1 97.9 2 95.6 0.4 99.4 3.4 99.4 % degradation Hydrocarbon Benzene Naphthenic acid Control Control Control without without without microbial With microbial microbial With microbial microbial With microbial Time (hrs.) consortia consortia consortia consortia consortia consortia 10 1.4 40.3 1.9 47.8 0 47.3 20 1.9 65.5 2.4 51.4 0 54.7 30 3.5 78.6 3.1 67.7 0 78.2 40 3.7 96.8 3.4 95.8 0.1 92.1 50 3.9 99.2 4.9 99.4 0.5 97.4

Example 4: Bio-Assisted Method Using with Two Reactors (with Free Form Haloalkaliphilic Bacterial Consortium)

[0104] Treatment is done in two continuously fed systems consisting of two CSTRs in series. The spent caustic is fed in the first reactor (2 L volume) along with nutrient system consisting of K.sub.2HPO.sub.4 (4 g/l), KH.sub.2PO.sub.4 (4 g/l), MgCl.sub.2 (0.2 g/l), 0.5 g/l of trace elements, sodium carbonate (5 g/l), yeast extract (7 g/l), ammonium nitrate (8 g/l), citrate (8 g/l), sorbitol ester (5 ppm), Oleic acid (230 ppm), pantothenic acid (20 ppm), thiamine (25 ppm).

[0105] The first reactor (2 L volume) was operated as 40° C. and inoculated with microbial consortium to obtain the cell count of >10.sup.2 CFU/ml and allowed to grow for 24 hrs. Subsequently, the spent caustic solution was continuously fed with HRT of 24 hrs.

[0106] The percentage oxygen saturation in the first reactor was maintained 20% maintain by sparging the air and it was stirred at 100 rpm. The effluent of first reactor was introduced in the second CSTR where the percentage of oxygen saturation level in the second reactor was maintained at 80% with stirring of 500 rpm. The second reactor was operated at ambient temperature and pressure. The HRT of the second reactor was 24 hrs.

[0107] To prevent the release of VOC from the system, the exhaust gas from the reactor was continuously recycled to the reactor. The recycled gas first passed a condenser to recover VOC and the fed to the same reactor. A control without microbes was run parallel. Un-treated and treated were analyzed for contaminant level using appropriate analytical tools. The results are shown in Table 4.

TABLE-US-00004 TABLE 4 Treatment of spent caustic with free form haloalkaliophilic bacterial consortium Content in % Control without After treatment with Haloalkaliphilic Haloalkaliphilic bacterial bacterial Contaminant consortium consortium Total sulfur 0.012 2.69 Sulphides 0.003 1.67 Mercaptans 0.017 2.62 Phenol 0.0002 0.020 Hydrocarbons 0.0001 0.32 Naphthenic acid 0.0002 0.024

Example 5: Bio-Assisted Method Using with Two Reactors (with Immobilized Haloalkaliphilic Bacterial Consortium)

[0108] Treatment is done in two continuously fed system consisting of two CSTRs in series as in the example-3 the consortia used in both CSTR was immobilized in porous glass and 10 ml (20 mg dry wt/ml porous glass) were used as inoculum. Table-5 showed that immobilized cells were equally effective as of free cells.

TABLE-US-00005 TABLE 5 Treatment of spent caustic with immobilized cells haloalkaliphilic bacterial consortium Content in % Control without After treatment with haloalkaliphilic haloalkaliphilic bacterial bacterial Contaminant consortium consortium Total sulfur 0.010 2.69 Sulphides 0.002 1.67 Mercaptans 0.018 2.62 Phenol 0.0001 0.020 Hydrocarbons 0.0002 0.32 Naphthenic acid 0.0001 0.024

Example 6: Bio-Assisted Method Using with Two Reactors (with Free Form Haloalkaliphilic Bacterial Consortium)

[0109] Treatment is done in two continuously fed systems consisting of two CSTRs in series. The spent caustic is fed in the first reactor (2 L volume) along with nutrient system consisting of K.sub.2HPO.sub.4 (4 g/l), KH.sub.2PO.sub.4 (4 g/l), MgCl.sub.2 (0.2 g/l), 0.5 g/l of trace elements, sodium carbonate (5 g/l), yeast extract (7 g/l), ammonium nitrate (8 g/l), citrate (8 g/l), sorbitol ester (5 ppm), Oleic acid (230 ppm), pantothenic acid (20 ppm), thiamine (25 ppm). The first reactor (2 L volume) was operated as 40° C. and inoculated with microbial consortium to obtain the cell count of >10.sup.2 CFU/ml and allowed to grow for 24 hrs. Subsequently, the spent caustic solution was continuously fed with HRT of 24 hrs. The percentage oxygen saturation in the first reactor was maintained 80% maintain by sparging the air and it was stirred at 1000 rpm. The effluent of first reactor was introduced in the second CSTR where the percentage of oxygen saturation level in the second reactor was maintained at 20% with stirring of 200 rpm. The second reactor was operated at ambient temperature and pressure. The HRT of the second reactor was 24 hrs. To prevent the release of VOC from the system, the exhaust gas from the reactor was continuously recycled to the reactor. The recycled gas first passed a condenser to recover VOC and the fed to the same reactor. A control without microbes was run parallel. Un-treated and treated were analyzed for contaminant level using appropriate analytical tools. The results are shown in table-5.

TABLE-US-00006 TABLE 6 Treatment of spent caustic with free form haloalkaliphilic bacterial consortium Content in % After treatment with Control without Contaminant microbial blend microbial blend Total sulfur 0.012 2.98 Sulphides 0.002 2.17 Mercaptans 0.019 3.69 Phenol 0.0002 0.050 Hydrocarbons 0.0001 0.67 Napthenic acid 0.0003 0.076

ADVANTAGES

[0110] The method of treatment of spent caustic of the present invention does not require efforts for maintaining critical parameters of pH as compared to that of the prior art wherein pH is maintained by addition of acids.

[0111] Further, treated spent caustic can be used for industrial applications by supplementing with minimum amount of solid metal hydroxide for treating oil and gas streams, maintenance of pH of ETP, paper and pulp industry etc.

[0112] The microbial consortia and its mediated process used in the present invention can degrade/transform multiple contaminants from the spent caustic in contrast to prior art wherein sulfides are treated in particular.

[0113] Also, said microbial consortia can be easily reproduced and does not require acclimatization.