Design of experiment methodology for Mannoprotein Industrial scale manufacturing Bioprocess: Optimization from a yeast cell factory

Kluyveromyces marxianus is a yeast species with various industrial applications. It's known for its ability to metabolize a wide range of substrates, including lactose, xylose, and cellobiose, which makes it useful in various biotechnological processes. In order to designing experiments (DOE) and design an optimized method for extracting and purification of mannoprotein using three homogenization, alkalinity, and bio-emulsifier extraction methods, as well as model data at three levels based on three factors: acidity, temperature, and the mannoprotein extraction method, this study looked at the growth of the yeast Kluyveromyces marxianus (IBRC-M 30114) in a 30-L fermentor to determine its optimized yield. Mannoprotein was extracted (in 27 runs with 3 parts) using QUALITEK-4 software and the Taguchi method. In the designed stage, various temperatures (25, 30, and 35 °C) and pHs (2, 5, and 7) were used in the homogenization, alkaline, and bio-emulsifier methods. The growth of the yeast K. marxianus in a fermenter (bioreactor) showed that the maximum biomass was obtained from the scale increase inside the reactor. This indicated that the maximum fermentation biomass 34.02 (g.l-1) was obtained from K. marxianus in 30 h at pH 4.6, 29 °C, 500 rpm, oxygen 19.7, inlet air volume 1 vvm3, and 36.6 DO. The maximum amount of mannoprotein 8.243 (mg 100 ml-1) from 10 (g.l-1) yeast biomass was extracted by the alkaline method with pH 5. In the bio-emulsifier method, the mannoprotein extraction was maximized at pH 7. The homogenization approach fared better than the alkaline method overall in terms of performance, while the alkaline method outperformed the other two methods homogenization and bioemulsifier in terms of volume or quantity. Mannoproteins have potential applications in drug delivery due to their biocompatibility and play important roles in various biological processes and have several applications in different indus.

Mannoprotein, K. marxianus, Optimization, Design of experiment (DOE), Bioprocess