Indian Journal of Biotechnology Vol 12, April 2013, pp 281-283 Biotransformation of low cost lignocellulosic substrates into vanillin by white rot fungus, Phanerochaete chrysosporium NCIM 1197 Bhushan Karode 1 *, Ulhas Patil 2 and Arpana Jobanputra 1 1 Department of Microbiology, PSGVPM’s ASC College, Shahada 425 409, India 2 Department of Microbiology, R C Patel ASC College, Shirpur 425 405, India Received 16 January 2012; revised 22 April 2012; accepted 10 June 2012 Vanillin (4-hydroxy-3-methoxybenzaldehyde), a pleasant smelling aromatic compound, occurs naturally in vanilla beans. It is widely used as a flavouring additive for beverages and aromatic additive for candles, fragrances, and perfumes. It is also noted as nutraceutical. Efforts have been made to obtain aromatic compounds produced by means of biological process, which employ microorganisms. White rot fungus, Phanerochaete chrysosporium NCIM 1197 was used to transform lignocellulosic substrates like rice husk, groundnut shell, etc. Further, detection and quantification of vanillin were carried out by TLC and spectrophotometrically. The maximum vanillin production (up to 55 μg/mL) was observed within 72 h using groundnut shell as a substrate. The use of various carbon sources, viz., glucose and starch, along with groundnut shell, affected vanillin production. Supplementation of glucose along with groundnut shell showed more increase in vanillin production (37 μg/mL within 96 h) in comparison to the supplementation of starch (up to 21 μg/mL within 96 h). Keywords: Biotransformation, ferulic acid, groundnut shell, rice husk, vanillin Vanillin (4-hydroxy-3-methoxybenzaldehyde), a pleasant smelling aromatic compound, occurs naturally in vanilla beans. It is one of the most important aromatic flavour compounds used in foods, beverages, perfumes and pharmaceuticals 1 . It is used as an aromatic additive for candles, incense, potpourri, fragrance and air fresheners. It is also noted as a nutraceutical because of its antioxidant and antimicrobial properties. The ice cream and chocolate industries together comprise 75% of the market for vanillin, where it is used as a flavoring agent, with smaller amounts being used in confectionaries and baked goods 2 . Natural vanillin is extracted from the seed pods of vanilla plant, an orchid native of Mexico. It represents less than 1% of the annual market demand 3 . Moreover, the demand for this natural ingredient is increasing day by day. This has led to the expensiveness of vanillin derived from plant. Various methods are being used to obtain vanillin from lignocellullosic substances. Alternative processes are being explored that use biotechnological methods involving fungi to produce vanillin 4 . Vanillin is the one of the known products of the degradation of lignin related substances by white rot fungi 5 . The aim of present investigation was to produce vanillin with the use of low cost lignocellulosic substrates, i.e., groundnut shell and rice husk using the basidiomycete, Phanerochaete chrysosporium. Several other substrates, viz., corn cobs (15%), fresh bagasse (18%), switch grass (12%) etc., have also been reported to contain a considerably good level of lignin 6,7 . However, the selection of groundnut shell was focused in the present investigation for the presence of high percentage of lignin (30-40%) and its easy availability vis-à-vis the other known substrates. P. chrysosporium NCIM 1197 on account of its potent lignin degrading ability was selected 8 . It was routinely cultivated on potato dextrose agar (PDA) solid medium at 28±1°C for 5 d. The same medium was also used for maintenance of the culture. The culture was stored on agar slants at 4°C. After every 30 d, the culture was transferred to fresh agar slants. Different agro-wastes, such as, groundnut shell and rice husk, were procured from local fields. Lignin, ferulic acid and vanillin were procured from Hi Media Ltd., Mumbai, India. All the chemicals used were of analytical grade. Loopful culture of P. chrysosporium NCIM 1197 was inoculated into 100 mL of PDB (potato dextrose broth). The culture was incubated at 28 o C and shaken at 120 rpm for 48 h for the production of inoculum. Then, 1% of the inoculum was transferred into 250 mL Erlenmeyer flasks containing 100 mL PDB providing one of the carbon sources, such as, lignin, —————— *Author for correspondence: Tel: +91-2565-229576; Fax: +91-2565-229576 (O) E-mail: arpana_j12@rediffmail.com