Optimization of medium using response surface methodology for l-DOPA production by Pseudomonas sp. SSA

• Published in: Biochemical engineering journal 2013-05, Vol.74, p.36-45
• Main Authors: Patil, Sushama A., Surwase, Shripad N., Jadhav, Shekhar B., Jadhav, Jyoti P.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biotechnology; copper sulfate; drugs; Fundamental and applied biological sciences. Psychology; gas chromatography-mass spectrometry; glucose; high performance liquid chromatography; HPTLC; L-dopa; Parkinson disease; peptones; Pseudomonas; Pseudomonas sp. SSA; Response surface methodology; screening; Tyrosinase; tyrosine
• ISSN: 1369-703X; 1873-295X
• DOI: 10.1016/j.bej.2013.02.021

•Optimization of nutritional parameters for bacterial l-DOPA production using Response surface methodology.•The Plackett–Burman design and Box–Behnken design was employed for optimization of media.•The maximum l-DOPA production obtained experimentally was found to be 3.251±0.12gl−1. l-3,4-Dihydroxyphenylalanine (l-DOPA) is the most potent drug used for treatment of Parkinsonism. Statistical optimization of nutritional parameters for the production of l-DOPA by Pseudomonas sp. SSA has been carried out using response surface methodology (RSM). Four most significant medium constituents identified by initial screening method of Plackett–Burman (PB) were glucose, peptone, l-tyrosine and CuSO4. In order to investigate quantitative effects of the four variables selected from PB design on l-DOPA production, Box–Behnken design was subsequently employed for further optimization. The medium having glucose 1.69gl−1, peptone 1.61gl−1, l-tyrosine 4.11gl−1 and CuSO4 0.03gl−1 was found to be optimum for maximum l-DOPA production. The optimization strategies used lead to a 6.27-fold increase in l-DOPA yield (3.251±0.12gl−1) compared to non-optimized medium (0.572±0.1gl−1). l-DOPA produced was further characterized by spectroscopic techniques, such as HPTLC, HPLC and GC–MS.