Chemistry: molecular biology and microbiology – Process of utilizing an enzyme or micro-organism to destroy... – Destruction of hazardous or toxic waste
Reexamination Certificate
2001-03-19
2003-09-02
Marx, Irene (Department: 1651)
Chemistry: molecular biology and microbiology
Process of utilizing an enzyme or micro-organism to destroy...
Destruction of hazardous or toxic waste
C435S254100, C435S911000
Reexamination Certificate
active
06613559
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to a process for the simultaneous decolorization and detoxification of molasses spent wash using strain of white-rot fungus, “
Flavodon flavus
” having accession No. NRRL 30302. The invention also relates to a novel strain “
Flavodon flavus
” which has been identified and which has been deposited at National Institute of Oceanography, Dona Paula, Goa, India, bearing the accession No. NIOCC #312 and also at Agricultural Research Service Culture Collection (ARS) [Patent collection (NRRL), USDA], 1815 North University Street, Peoria, Ill. 61604, U.S.A., bearing accession No. NRRL 30302 on Mar. 10, 2000.
BACKGROUND OF THE INVENTION
Distilleries in India use sugarcane molasses as the raw material for production of alcohol. The effluents from such distilleries contain large amounts of molasses spent wash (MSW). Molasses spent wash from such plants contain high Biological oxygen demand (BOD) and chemical oxygen demand (COD) and suspended solids. MSW pollutes aquatic ecosystems due to its intense brown color which cuts off light, prevents photosynthesis and causes anaerobic conditions. Next to effluent from paper and pulp mill and tannery, molasses spent wash is a major environmental hazard to land or aquatic sources where they are discharged. Due to the importance attached to prevention of environmental pollution, environmental agencies all over the world are imposing strict regulations for mitigation of pollution from industries. The effluents from distilleries containing colored pigments, high-suspended solids, a high concentration of BOD and COD, besides causing aesthetic damage to sites, are toxic to resident flora and fauna. Wastewater from fermentation plant using sugarcane molasses contains a large amount of a dark brown pigment called molasses melonoidin which is not broken down by usual biological treatments. Melanoidin pigments are the products of “Maillard reaction” between sugars and amino compounds produced on heating (Wedzicha and Kaputo, 1992). Anaerobic digestion of MSW-containing effluents is one of the treatments followed by distilleries and the resulting dark brown sludge is used as a fertilizer. The effluent after such treatment has reduced Chemical Oxygen Demand (COD) and Biological Oxygen Demand (BOD) but is still dark brown in color and is a major problem with distilleries. Removal of such colored pigments in molasses spent wash is termed bioremediation. Color removal in such effluents using terrestrial fungi has been reported (Sirianantapiboon et al. 1988).
Normally, the molasses spent wash wastewater disposal includes physical-chemical treatment, waste-minimization and biological treatment. Biological treatment includes pretreatment with activated sludge of spent wash and treatment in stabilization ponds. Unfortunately, wastewater treatment facilities are often unable to completely remove melanoidin pigments from wastewater and thus contribute to pollution of soil and aquatic habitats. They are even proven to be toxic to aquatic animals (Fitzgibbon et al. 1995). Hence, there is a need for developing an efficient method for the treatment of the molasses spent wash.
PRIOR ART RELATING TO THE INVENTION
Various organisms have been tried for degradation of molasses spent wash in distillery wastewater and bioremediation.
(i) A reference may be made to a publication wherein filamentous fungi of the order
Mycelia Sterilia
were screened for decolorization of molasses pigments (Sirianuntapiboon, et al. 1988). The maximum decolorization was achieved within 7-8 days.
(ii) A reference may be made to a publication wherein a thermophilic strain of
Aspergillus fumigatus
G-2-6 was shown to decolorize molasses melanoidin to an extent of 75% at 45° C. within 3 days with shaking (Ohmomo et al., 1987). Shaking of culture and maintaining higher temperature are two additional requirements and thus have practical limitations on a commercial scale.
(iii) A reference may be made to a publication wherein Artemia larvae have been used for treatment of distillery waste and have shown to decrease B.O.D and C.O.D levels considerably. However, they do not help in decolorizing the molasses spent wash (Rahaman, et al., 1992).
(iv) A reference may be made to a publication wherein the melonoidins prepared from a glucose-glycine system are decolorized and degraded on ozone treatment. (Kim and Park 1986). However, this process is expensive and not practical.
(v) A reference may be made to a publication wherein about 85% color from molasses spent wash (MSW) was removed after 10 days by the white-rot fungus
Phanerochaete chrysosporium
grown in a medium containing 6.25% MSW (Fahy et al. 1997). However,
Flavodon flavus,
strain NIOCC 312 used in our studies removes 90% color after 10 days in a medium containing 10% MSW.
(vi) A reference may be made to a publication wherein strain Ps4a of
Trametes versicolor,
a white-rot fungus, yielded about 79% color removal of molasses pigment by day 4 (Aoshima et al. 1985) in a medium containing 10% melanoidin pigments. However, they have used wastewater treated with activated sludge and not raw molasses spent wash from a distillery as the applicants have done in their studies. Moreover, the applicant's isolate is capable of growth and decolorization of MSW in the presence of sea salts.
(vii) A reference also may be made to the applicant's own co-pending U.S. patent application Ser. No. 09/456,559 titled “Novel white rot-lignin-modifying fungus
Flavadon flavus
and a process for removing dye from dye containing water or soil using the fungus” filed on Dec. 7, 1999, wherein the novel fungus “
Flavadon flavus
” has been isolated, identified and was used for the decolorization of the effluents from the textile, leather and paper industries, containing synthetic dyes comprising azo, heterocyclic and polymeric dyes.
The applicants (C.S.I.R.) in their co-pending U.S. patent application Ser. No. 09/456,559 have found out that the fungal isolate
F. flavus
produces lignin-modifying enzymes such as manganese-dependent peroxidase, lignin peroxidase and laccase in conventional natural and synthetic media prepared with distilled water as well as half-strength artificial sea water and also in powdered sugarcane bagasse, pine and spruce wood-powder media. By virtue of these lignin-modifying enzymes which break down a broad range of xenobiotics, this fungus is useful in 80 to 100 percent decolorization of effluents from paper mill, leather industry and wide range of synthetic dyes such as azo dyes, heterocyclic and polymeric dyes within 4 to 6 days. The dyes specifically tested were Azure B, Brilliant green, Crystal violet, Congo red, Remazol Brilliant blue R, Poly B-411 and Poly R-478at 0.02 percent concentration.
The previous application also described growth of the said fungus on a large scale using inexpensive raw materials such as sugarcane baggase suspended in distilled water or half strength artificial sea water. The biomass of the fungus thus obtained can be used for seeding soil contaminated with synthetic dyes. The above study was restricted to decolorization of effluents from paper mills and leather industries and the present application is for the new field of simultaneous decolorization and detoxification of molasses spent wash, which has not been identified so far.
OBJECTS OF THE INVENTION
The main object of the present invention is to develop a method for the simultaneous decolorization and detoxification of molasses spent wash using a novel white-rot lignin modifying fungus strain “
Flavodon flavus
”, which has been deposited at National Institute of Oceanography, Dona Paula, Goa, India, bearing the accession No. NIOCC #312 and also at ARS Patent Collection (NRRL), USDA, Illinois, U.S.A., bearing accession No. NRRL 30302.
SUMMARY OF THE INVENTION
To meet the above object, the present invention provides process for the simultaneous decolorization and detoxification of the molasses spent wash using a white rot lignin modifying fungus strain “
Flavodon flavus
”, which has been depo
Raghukumar Chandralata
Shailaja Mysore Srinivasamurthy
Council of Scientific and Industrial Research
Marx Irene
Smith , Gambrell & Russell, LLP
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