Catalysis Letters

, Volume 148, Issue 2, pp 712–724 | Cite as

A Statistical Approach to Optimize Cold Active β-Galactosidase Production by an Arctic Sediment Pscychrotrophic Bacteria, Enterobacter ludwigii (MCC 3423) in Cheese Whey

  • Aneesa P. Alikunju
  • Susan Joy
  • Mujeeb Rahiman
  • Emilda Rosmine
  • Ally C. Antony
  • Solly Solomon
  • K. Manjusha
  • A. V. Saramma
  • K. P. Krishnan
  • A. A. Mohamed Hatha


Cold active β-galactosidases which catalyze lactose hydrolysis and transglycosylation reactions at low temperature make them highly potential biocatalyst in biotechnology, pharmaceutical and food processing industries. Moreover, an interest towards the utilization of diary industrial waste, whey and its constituents, for manufacturing a wide range of valuable products at reliable cost is increasing among researchers in order to facilitate its wider commercial use. In the present study, the fermentation parameters for the maximum production of cold active β-galactosidase from a psychrotrophic bacterium, Enterobacter ludwigii in cheese whey was optimized by exploring statistical methods, Plackett-Burman design (PBD) and central composite design (CCD). Three most significant factors viz, pH, whey and tryptone out of 11 were selected by PBD and were further optimized by response surface methodology using CCD. The optimal levels of pH, whey and tryptone were indicated as 7.3, 82 (v/v) % and 3.84 g% respectively. An overall 3.6-fold increase in cold active β-galactosidase production (34.37 U/mL) was achieved in optimized medium compared to the yield from unoptimized medium. The quadratic regression model was proven to be adequate (p = 0.0001, R 2  = 0.9880, CV = 7.96%) and the response (cold active β-galactosidase production) obtained on validation coincident with the predicted value.

Graphical Abstract


Arctic Cold active β-galactosidase Response surface methodology Whey 



The authors are thankful to UGC-BSR, India for funding the research work (UGC Grant No. F.No. 25-1/2014-15(BSR)/5-24/2007/(BSR). Authors are also thankful to the Department of Marine Biology, Microbiology and Biochemistry and Sophisticated Testing and Instrumentation Centre (STIC) at Cochin University of Science and Technology (CUSAT) and National Centre for Antarctic and Ocean Research (NCAOR) for providing the facilities to carry out the research. Authors acknowledge Dr. C. K. Radhakrishnan and K. T. Thomas for their valuable suggestions and support.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest in the publication.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  • Aneesa P. Alikunju
    • 1
  • Susan Joy
    • 1
  • Mujeeb Rahiman
    • 2
  • Emilda Rosmine
    • 1
  • Ally C. Antony
    • 1
  • Solly Solomon
    • 1
  • K. Manjusha
    • 3
  • A. V. Saramma
    • 1
  • K. P. Krishnan
    • 4
  • A. A. Mohamed Hatha
    • 1
  1. 1.Department of Marine Biology, Microbiology and BiochemistryCochin University of Science and TechnologyCochinIndia
  2. 2.Department of Aquaculture and Fishery MicrobiologyMES Ponnani CollegePonnaniIndia
  3. 3.Department of MicrobiologySt.Xaviers College for WomenAluvaIndia
  4. 4.National Center for Antarctic and Ocean ResearchGoaIndia

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