ORIGINAL PAPER Mixture design as a ﬁrst step for optimization of fermentation medium for cutinase production from Colletotrichum lindemuthianum Fred J. Rispoli Æ Vishal Shah Received: 21 August 2006 / Accepted: 28 December 2006 / Published online: 6 February 2007 Ó Society for Industrial Microbiology 2007 Abstract Cutinase enzymes from fungi have found diverse applications in industry. However, most of the available literature on cutinase production is related to the cultivation of genetically engineered bacteria or yeast cells. In the present study, we use mixture design experiments to evaluate the inﬂuence of six nutrient elements on production of cutinase from the fungus Colletotrichum lindemuthianum. The nutritional ele- ments were starch, glucose, ammonium sulfate, yeast extract, magnesium sulfate, and potassium phosphate. In the experimental design, we imposed the constraints that exactly one factor must be omitted in each set of experiments and no factor can account for more than one third of the mixture. Thirty different sets of experiments were designed. Results obtained showed that while starch is found to have negative inﬂuence on the production of the enzyme, yeast extract and potassium phosphate have a strong positive inﬂuence. Magnesium sulfate, ammonium sulfate, and glucose have low positive inﬂuence on the enzyme production. Contour plots have also been created to obtain infor- mation concerning the interaction effects of the media components on enzyme production. Keywords Cutinase  Colletotrichum lindemuthianum  Media optimization  Simplex lattice  Mixture design Introduction The biotechnology industry is developing novel prod- ucts in a wide range of areas such as human and animal healthcare, agriculture, environment and diagnostics. Modern tools like genetic engineering, computational analysis and computer simulation have helped to de- velop biological systems producing new products at a fast rate. These products vary from whole cells to extracellular secondary metabolites. However, as new systems are being developed by scientists in laborato- ries, engineers are faced with the task of developing the optimal media conditions for culture growth to ensure high yields and product quality. Culture med- ium optimization is a critical step in the fermentation process development and often continues throughout the production life of the fermentation product. The process of developing an optimum medium for maximum production involves a stage where the crit- ical medium components and process parameters inﬂuencing production of the desired product are screened. The primary goal in this step is to study the statistical signiﬁcance of an effect that a particular factor exerts on the dependent variable of interest. Once the components critical to the production are screened, the second stage of media optimization is to ﬁnd the optimum concentration of each component for maximum product formation. While developing an industrial process it is imperative to carry out the optimization studies that can be scaled up at larger scale with ease. Screening the process parameters such as nutrition components for carbon, nitrogen, phos- phorus and trace elements; physico-chemical parame- ters such as pH, temperature, and aeration rate; both involve determining which parameters have F. J. Rispoli Department of Mathematics, Dowling College, Idle Hour Blvd, Oakdale, NY 11955, USA V. Shah (&) Department of Biology, Dowling College, Idle Hour Blvd, Oakdale, NY 11955, USA e-mail: ShahV@dowling.edu 123 J Ind Microbiol Biotechnol (2007) 34:349–355 DOI 10.1007/s10295-007-0203-y Downloaded from https://academic.oup.com/jimb/article/34/5/349/5993039 by guest on 05 March 2024