OPTIMIZATION OF THERMOPHILIC PULLULANASE AND α-AMYLASE PRODUCTION BY AMYLOLYTIC YEAST

D.S. DJEKRIF¹*, L. GILLMANN², N. COCHET³, L. BENNAMOUN⁴, A. AIT-KAKI⁵, K. LABBANI⁶, T. NOUADRI⁷, Z. MERAIHI^8
1Department of Biochemistry, Faculty of Biology, Research lab in Microbiology Engineering, Biopole Chaab Erssas, Constantine University 1, 25000, Constantine, Algeria.
²SONAS-IUT laboratory, University of Angers, 49016, Angers, France.
³Research lab. Transformations in Integrated Renewable Material, University of Technology, 60205, Compiègne, Compiègne Cedex, France.
⁴Department of Biochemistry, Faculty of Biology, Research lab in Microbiology Engineering, Biopole Chaab Erssas, Constantine University 1, 25000, Constantine, Algeria.
⁵Department of Biochemistry, Faculty of Biology, Research lab in Microbiology Engineering, Biopole Chaab Erssas, Constantine University 1, 25000, Constantine, Algeria.
⁶Department of Biochemistry, Faculty of Biology, Research lab in Microbiology Engineering, Biopole Chaab Erssas, Constantine University 1, 25000, Constantine, Algeria.
⁷Department of Biochemistry, Faculty of Biology, Research lab in Microbiology Engineering, Biopole Chaab Erssas, Constantine University 1, 25000, Constantine, Algeria.
⁸Department of Biochemistry, Faculty of Biology, Research lab in Microbiology Engineering, Biopole Chaab Erssas, Constantine University 1, 25000, Constantine, Algeria.
* Corresponding Author : scheherazad2002@hotmail.com

Received : 23-08-2014     Accepted : 06-09-2014     Published : 11-12-2014
Volume : 6     Issue : 2       Pages : 559 - 569
Int J Microbiol Res 6.2 (2014):559-569

Keywords : Thermophilic enzymes, Clavispora lusitaniae, Plackett-Burman design, CCD, optimization.
Academic Editor : Jiong Hong, Elena Bartkiene, Maria Balcerek, Bousba Ratiba
Conflict of Interest : None declared

The amylolytic enzymes are the most important group of commercially produced enzymes such as α-amylase (EC 3.2.1.1) and debranching pullulanase (pullulan 6-glucanohydrolase; EC 3.2.1.41). In the present study, optimization of physical and nutritional parameters influencing pullulanase and α-amylase production attempted using the response surface methodology (RSM) from a strain of Clavispora lusitaniae ABS7 isolated from arid zone wheat seeds (Algerian Sahara). A Plackett-Burman design was used for screening of critical components, while the optimum selected factors are determined by the Wilson-Box design. Eight variables (agitation, pH, temperature, starch, yeast extract, (NH4)2SO4, salts and trace-elements) were studied with the screening test. The results revealed that six factors had greater influence on the amylopullulanase production. The CCD was then used to determine the maximum amylopullulanase concentration. The optimum conditions for the highest α-amylase production (13456,36 ± 300 U) and pullulanase production (12611,6±154 were as follows: agitation 136,56 rpm, temperature 54,14°C, starch 2,66 g/l, yeast extract 0,365 g/l, salts solution 8,75 ml/l and trace-elements solution 4,3 ml/l. Also the strain grows at the optimum condition, in fermenter and produces maximally amylopullulanase on an alkaline medium.