Abstract
BackgroundPichia guilliermondii was found capable of expressing the recombinant thermostable lipase without methanol under the control of methanol dependent alcohol oxidase 1 promoter (AOXp 1). In this study, statistical approaches were employed for the screening and optimisation of physical conditions for T1 lipase production in P. guilliermondii.ResultThe screening of six physical conditions by Plackett-Burman Design has identified pH, inoculum size and incubation time as exerting significant effects on lipase production. These three conditions were further optimised using, Box-Behnken Design of Response Surface Methodology, which predicted an optimum medium comprising pH 6, 24 h incubation time and 2% inoculum size. T1 lipase activity of 2.0 U/mL was produced with a biomass of OD600 23.0.ConclusionThe process of using RSM for optimisation yielded a 3-fold increase of T1 lipase over medium before optimisation. Therefore, this result has proven that T1 lipase can be produced at a higher yield in P. guilliermondii.
Highlights
Pichia guilliermondii was found capable of expressing the recombinant thermostable lipase without methanol under the control of methanol dependent alcohol oxidase 1 promoter (AOXp 1)
Some lipase producing yeasts belonged to Pichia, Kluyveromyces, Candida and Torulaspora genre are methylotrophic in nature [7,8,9]
Effects of different media on T1 lipase production Alterations in the pH of a cultivation medium can lead to a change in the charge of a medium, which affects cell growth and recombinant protein production. pH is mentioned as a crucial fermentation factor affecting cell growth and recombinant proteins production if not controlled [26,27,28]
Summary
Pichia guilliermondii was found capable of expressing the recombinant thermostable lipase without methanol under the control of methanol dependent alcohol oxidase 1 promoter (AOXp 1). Microbial thermostable lipases are known to be originated from bacterial, fungi and yeast with many applications in industrial and biotechnological processes [4,5,6]. Some lipase producing yeasts belonged to Pichia, Kluyveromyces, Candida and Torulaspora genre are methylotrophic in nature [7,8,9]. They metabolise methanol as their carbon and energy source [10,11,12].
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