Title

Metabolic pathway reconstruction of eugenol to vanillin bioconversion in Aspergillus niger

Authors

Suchita Srivastava¹, Suaib Luqman¹, Feroz Khan², Chandan S. Chanotiya³, Mahendra P. Darokar ^1*

Affiliation

¹Molecular Bioprospection Division, ²Metabolic & Structural Biology Division, ³Chemical Sciences Division, Central Institute of Medicinal & Aromatic Plants, (Council of Scientific & Industrial Research), Lucknow-226015 (UP) INDIA

Email

mpdarokar@yahoo.com

Article Type

Hypothesis

Date

Received November 01, 2009; Revised November 18, 2009; Accepted December 13, 2009; Published January 23, 2010

Abstract

Identification of missing genes or proteins participating in the metabolic pathways as enzymes are of great interest. One such class of pathway is involved in the eugenol to vanillin bioconversion. Our goal is to develop an integral approach for identifying the topology of a reference or known pathway in other organism. We successfully identify the missing enzymes and then reconstruct the vanillin biosynthetic pathway in Aspergillus niger. The procedure combines enzyme sequence similarity searched through BLAST homology search and orthologs detection through COG & KEGG databases. Conservation of protein domains and motifs was searched through CDD, PFAM & PROSITE databases. Predictions regarding how proteins act in pathway were validated experimentally and also compared with reported data. The bioconversion of vanillin was screened on UV-TLC plates and later confirmed through GC and GC-MS techniques. We applied a procedure for identifying missing enzymes on the basis of conserved functional motifs and later reconstruct the metabolic pathway in target organism. Using the vanillin biosynthetic pathway of Pseudomonas fluorescens as a case study, we indicate how this approach can be used to reconstruct the reference pathway in A. niger and later results were experimentally validated through chromatography and spectroscopy techniques.

Keywords

Metabolic Pathway, reconstruction, eugenol, vanillin, Pseudomonas, Aspergillus, GC-MS.

Citation

Srivastava et al., Bioinformation, 4 (7) 320-325, (2010)

Edited by

P. Kangueane

ISSN

0973-2063

Publisher

Biomedical Informatics

License

This is an Open Access article which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. This is distributed under the terms of the Creative Commons Attribution License.