JP2000063499 | HEAT RESISTANT RESIN AND ITS PREPARATION |
JPS63192392 | PRODUCTION OF PLANT SECONDARY METABOLITE |
JPH04311394 | PREPARATION OF PYROCATECHOLIC COMPOUND |
LALI ARVIND M (IN)
YEOLE MAHENDRA M (IN)
LALI ARVIND M (IN)
AMENDED CLAIMS received by the International Bureau on 14 July 2014 (14.07.2014) I/We claim: 1. A method for continuous biotransformation of the substituted aryl carboxylic acids to their corresponding substituted aryl aldehydes of substituted aryl alcohols with high purity and yield comprising: a) preparing the spore suspension of the culture Pycnoporus cinnabarinus (NCIM- 181); b) culturing the fungus Pycnoporus cinnabarinus (NCIM-1181) from the spore suspension in a fermentor containing culture medium of pH in the range of 3-6 for 48-96 hrs to obtain cell pellets for biotransformation; c) adding substituted aryl carboxylic acid to the fermentor of step (b) to obtain the final concentration of 1-3 g/L of the substituted aryl carboxylic acid and allowing the biotransformation to be carried out for a time period sufficient to obtain substituted aryl aldehyde or substituted aryl alcohol in the fermentation broth; d) filtering the fermentation broth to obtain a permeate fraction comprising substituted aryl aldehyde or substituted aryl alcohol, and a retentate fraction comprising culture cells and/or spores and/or mycelia and/or pellets; wherein the retentate fraction is recycled to the fermentor; e) passing the permeate fraction through capture column for selective adsorption of the substituted aryl aldehyde or substituted aryl alcohol obtained in step (d) at pH in the range of 6-8; and recycling the remaining permeate fraction to the fermentor; f) washing the capture column with 0.1 M NaHC03 solution to remove unreacted substituted aryl carboxylic acid; g) eluting the capture column with an organic solvent to obtain an eluent containing substituted aryl aldehyde or substituted aryl alcohol ; h) concentrating the eluent containing substituted aryl aldehyde or substituted aryl alcohol by vacuum distillation at temperature of 60°C and allowing to cool down gradually for 6-12 hrs at temperature in the range of 10-30°C to obtain a crude substituted aryl aldehyde or substituted aryl alcohol; wherein the crude substituted aryl aldehyde or substituted aryl alcohol having purity in the range of 86-92%; and i) crystallizing the crude substituted aryl aldehyde or substituted aryl alcohol from step (h) to obtain a pure substituted aryl aldehyde or substituted aryl alcohol; wherein the pure substituted aryl aldehyde or substituted aryl alcohol having purity in the range of 96-99.5% and yield in the range of 84-93%. 2. The method as claimed in claim 1 , wherein substituted aryl carboxylic acid is selected from the group comprising of 4-hydroxy 3-methoxy benzoic acid, 3- hydroxy-4-methoxy benzoic acid, p-Hydroxy benzoic acid, 3-Amino-5- chlorobenzoic acid, 3,5-Dimethoxy-4-hydroxybenzoic acid. 3. The method as claimed in claim 1 , wherein obtainable substituted aryl aldehyde is from the group comprising of 4-Hydroxy-3-methoxybenzaldehyde, 3-Hydroxy-4-ethoxybenzaldehyde, p-hydroxy benzaldehyde, 3, 5-Dimethoxy-4- hydroxybenzaldehyde, 3-Amino-5-chlorobenzaldehyde. 4. The method as claimed in claim 1, wherein obtainable substituted aryl alcohol is from the group comprising of 4-hydroxy-3-methoxybenzyl alcohol, p- hydroxy benzyl alcohol, 3-Amino-5-chlorobenzyl alcohol, 3-Hydroxy-4- methoxybenzyl alcohol. 5. The method as claimed in claim 1 , wherein substituted aryl carboxylic acid is added in its salt form. 6. The method as claimed as in claim 1 , wherein biotransformation is carried out in fed batch or continuous manner. 7. The method as claimed in claim 1 , wherein filtration is carried out using membrane filter or perforated plates, having pore size atleast 0.22 micron. 8. The method as claimed in claim 1, wherein capture column is packed with hydrophobic adsorbent. 9. The method as claimed in claim 8, wherein hydrophobic adsorbent used in capture column is polystyrene divinyl benzene based or any polystyrene methacrylate based polymeric adsorptive resin. 10. The method as claimed in claim 1 , wherein organic polar solvent is selected from the group consisting of methanol, ethanol, isopropanol, butanol, water, acetone, ethyl acetate, most preferably methanol. 11. The method as claimed in claim 1, wherein the crystallization is done in distilled water or potable water. 12. The method as claimed in claim 1, wherein the time period sufficient to carry out the biotransformation is in the range of 50-90 hrs. |
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