A NEWLY ISOLATED YEAST AS AN EXPRESSION HOST FOR RECOMBINANT LIPASE

作者：Oslan Siti Nurbaya; Salleh Abu Bakar^*; Abd Rahman Raja Noor Zaliha Raja; Leow Thean Chor; Sukamat Hafizah; Basri Mahiran

来源：Cellular and Molecular Biology Letters, 2015, 20(2): 279-293.

DOI：10.1515/cmble-2015-0015

• Pichia guilliermondii
• Expression
• Thermostable lipase
• Alcohol oxidase
• Geobacillus zalihae
• Yeast
• Pichia pastoris
• His-tagged fusion protein
• Methylotrophic yeast
• Local host

Pichia guilliermondii strain SO isolated from spoiled orange was developed for use as an alternative expression host by using Pichia pastoris as the model of the experiment. This is the first study to report on the capability of P. guilliermondii SO as a host to express thermostable T1 lipase from Geobacillus zalihae. Alcohol oxidase and formaldehyde dehydrogenase promoters were present in the yeast genome. Interestingly, the recombinant yeast [SO/pPICZaB/T1-2 (SO2)] took only 30 h to reach optimal production with minimal methanol induction [1.5% (v/v)] in YPTM medium, as compared to P. pastoris, which took longer to reach its optimal condition. The purification yield of the His-tagged fusion lipase was 68.58%, with specific activity of 194.58 U/mg. The optimum temperature was 65 degrees C at pH 9 in glycine-NaOH buffer, and it was stable up to 70 degrees C in a wide pH range from pH 5 to 12. In conclusion, a newly isolated yeast from spoiled orange has been proven suitable for use as an expression host.

• 出版日期2015-6