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• W3149703706 abstract "Lipases are hydrolytic enzymes, ranked as third most relevant industrial enzymes with5 % share in global enzyme market. Thermostable T1 lipase from Geobacillus zalihaewas previously expressed under the regulatory control of alcohol oxidase promoter 1(AOXp 1) in the methylotrophic yeast Pichia guilliermondii, isolated from spoiledorange. Methanol was found to be a vital compound to induce the promoter activity inPichia pastoris. In this study P. guilliermondii has shown the potential to express therecombinant lipase without methanol under the regulation of AOXp 1. This studysought to optimise medium conditions of thermostable lipase with and withoutmethanol as inducer. The expression of T1 lipase without methanol was expected toreduce cost and toxicity effect of methanol.Buffered and non-buffered media compositions were studied for T1 lipase production,the media were first, supplemented with methanol then without methanol. Buffercomplex methanol medium was observed to be optimum for T1 lipase production witha 3-fold increase over non-buffered methanol medium. One-factor-at-a-time,conventional method of optimisation was used to identify significant data range formedium parameters. Using the observed data range, eight parameters which includestemperature, pH, inoculum size, biomass concentration, incubation time, shakingspeed, culture volume and methanol concentration, were screened for lipaseproduction in methanol medium using Plackett-Burman Design. Temperature,inoculum size, culture volume and incubation time, were observed to exert significanteffect on lipase production. These parameters were optimised using Box-BehnkenDesign of Response Surface Methodology. Optimum levels of these parameters werepredicted at temperature 34 oC, culture volume 190 mL, inoculum size 4 v/v andincubation time 24 h with an experimented lipase activity of 9.26 U/mL. Over 2-foldincrease before optimization in methanol medium was observed and 6-fold increaseover previous research work. On the other hand, six parameters which include temperature, pH, inoculum size,incubation time, shaking speed and culture volume were screened for T1 lipaseproduction in medium without methanol. Three parameters were observed to havesignificant effect on lipase production, then, these parameters were further optimisedusing Box-Behnken Design and their optimum levels were achieved at pH of 6,inoculum size 2 v/v and incubation time of 24 h as was predicted and the experimentedlipase activity of 2.012 U/mL was observed. This result gave 4-fold increase overlipase production before optimisation in medium without methanol. Recombinant T1lipase production was further scaled up to 3 L in the bioreactor for 128 h in methanolmedium and lipase activity observed was 12 U/mL at 120 h incubation time.Meanwhile, scale up in medium without methanol yielded a lipase activity of 3.5U/mL at 118 h incubation time. The result has proven that, the time taken for optimumlipase production without methanol was faster from the production with methanol inthe bioreactor.In conclusion, temperature, pH, inoculum size, incubation time and culture volume forrecombinant T1 lipase production were optimised in shake-flask and applied to thebioreactor level. Lipase was also produced in medium without methanol both at theshake flask level and the bioreactor. Thus, medium parameters optimisation hasproven to be very useful in enhancing thermostable lipase production in recombinantP. guilliermondii. Although T1 lipase has been expressed in medium without methanolbut it is lower compared to medium with methanol." @default.
• W3149703706 created "2021-04-13" @default.
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• W3149703706 date "2017-07-01" @default.
• W3149703706 modified "2023-09-26" @default.
• W3149703706 title "Enhancing recombinant T1 lipase production in Pichia guilliermondii" @default.
• W3149703706 hasPublicationYear "2017" @default.
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