Screening and identification of microorganisms for decolorization of molasses spent wash

Microorganisms were screened from the natural environment for decolorization of molasses spent wash,and the isolated strains were then employed in the treatment of actual wastewater.The primary screening was carried out on agar plates supplemented with synthesized melanoidin as the target substrate,since melanoidin is one of the most refractory pigments in wastewater.Promising microorganisms were further selected through secondary screening by decolorization of untreated actual wastewater in shaking flask cultures.Gel filtration chromatography was used to determine the molecular weight distribution of pigments in molasses spent wash before and after decolorization.A strain named A5P1 was isolated from the soil samples collected,showing a good ability of decolorizing molasses spent wash,and was later identified as Aspergillus flavus by morphology and ITS sequence analysis.Experimental study of factors affecting the decolorization performance of strain A5P1 gave the optimal conditions as follows: 4.3×104 mL-1 of inoculum size,medium with initial pH of 4.5 and cultivation at 39℃.It could decolorize 53.0% of the pigments in the untreated molasses spent wash and decreased 80% of chemical oxygen demand after four-day incubation.The result of gel filtration chromatography demonstrated that both the large and small molecular weight fractions of pigments in the molasses spent wash could be removed by strain A5P1.Based on the measurement of enzyme activities,at least three different kinds of enzymes,i.e.the enzyme with H2O2-producing activity,laccase and manganese peroxidase were involved in the decolorization process.Therefore,the decolorization mechanism of strain A5P1 was preliminarily considered to be mainly biodegradation,with bioadsorption as a minor reaction.