Journal of Industrial Microbiology & Biotechnology

, Volume 38, Issue 9, pp 1407–1417 | Cite as

High level phytase production by Aspergillus niger NCIM 563 in solid state culture: response surface optimization, up-scaling, and its partial characterization

  • K. Bhavsar
  • V. Ravi Kumar
  • J. M. KhireEmail author
Original Paper


Phytase production by Aspergillus niger NCIM 563 was optimized by using wheat bran in solid state fermentation (SSF). An integrated statistical optimization approach involving the combination of Placket–Burman design (PBD) and Box–Behnken design (BBD) was employed. PBD was used to evaluate the effect of 11 variables related to phytase production, and five statistically significant variables, namely, glucose, dextrin, NaNO3, distilled water, and MgSO4·7H2O, were selected for further optimization studies. The levels of five variables for maximum phytase production were determined by a BBD. Phytase production improved from 50 IU/g dry moldy bran (DMB) to 154 IU/g DMB indicating 3.08-fold increase after optimization. A simultaneous reduction in fermentation time from 7 to 4 days shows a high productivity of 38,500 IU/kg/day. Scaling up the process in trays gave reproducible phytase production overcoming industrial constraints of practicability and economics. The culture extract also had 133.2, 41.58, and 310.34 IU/g DMB of xylanase, cellulase, and amylase activities, respectively. The partially purified phytase was optimally active at 55°C and pH 6.0. The enzyme retained ca. 75% activity over a wide pH range 2.0–9.5. It also released more inorganic phosphorus from soybean meal in a broad pH range from 2.5 to 6.5 under emulated gastric conditions. Molecular weight of phytase on Sephacryl S-200 was approximately 87 kDa. The K m and V max observed were 0.156 mM and 220 μm/min/mg. The SSF phytase from A. niger NCIM 563 offers an economical production capability and its wide pH stability shows its suitability for use in poultry feed.


Phytase Aspergillus niger Solid state fermentation Statistical methods Response surface optimization 



One of the authors, Ms Kavita Bhavsar, thanks Council of Scientific and Industrial Research, Government of India for the financial assistance. We also gratefully acknowledge support and facilities provided by the Center of Excellence in Scientific Computing, National Chemical Laboratory, India.


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Copyright information

© Society for Industrial Microbiology 2010

Authors and Affiliations

  1. 1.NCIM Resource Center, National Chemical LaboratoryPuneIndia
  2. 2.Chemical Engineering and Process Development DivisionNational Chemical LaboratoryPuneIndia

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