I try to find yeast H. polymorpha that have auxotrophic markers in many collections for use with my M.Sc. project, but I found only H. polymorpha NCYC495 ura3- or leu2- available.
I read many books and publications which explain that for recombinant protein expression. A yeast strain that derived from CBS4732 or DL-1 are more proper and generally used for recombinat protein expression because strain CBS4732 and DL-1 can grow well in methanol as a C-source, but NCYC495 can not grow well in methanol.
Methanol is used for the induction of MOX or FMD promoters (these promoters are derived from the methanol utilization pathway). So, the author concluded that strain NCYC495 is not good for recombinant protein expression.
(I guess because these promoters, if used with strain NCYC495 cannot be strong and tightly regulated promoters like in the CBS4732 or DL-1 strain. But my assumption may be correct or incorrect because I can't find any publication to verify my assumption.)
My questions are:
1. If I can get only this strain, is it possible to use the FMD promoter with NCYC495 strain for recombinant protein production?
2. Since NCYC495 cannot grow well in methanol, then can MOX or FMD promoters tightly regulate and be strongly induced by methanol or not?
Thank you,
Tatpong.
Hello Tatpong,
H. polymorpha NCYC495 is used for the production of recombinant proteins (I added a paper in which it was used for the production of the heat shock protein gp96). Furthermore, the alcohol oxidase promoter is strongly induced when the cells are shifted to MeOH-containing medium. Growth of NCYC495 in methanol medium can be supported by the addition of 0.05% glycerol to the medium.
Regarding question 1: In my opinion you could use the strain. I would probably choose the alcohol oxidase promoter instead of FMD, however.
Regarding question 2: The AOX promoter usually gives good results regarding tight regulation and strong induction.
Best regards.
I fully agree with Christian and so the NCYC495 is an excellent strain for the production of recombinant proteins. If you need any vectors to overproduce your protein of interest please feel free to contact me. We are having a good collection of plasmid which are suitable for overproduction. You could also consider to clen your gene of interest behind two promoters namely the AOX promoter to overexpress on methanol and the ADH1 promoter to overexpress on glucose.
Novel genetic tools for Hansenula polymorpha.
Saraya R, Krikken AM, Kiel JA, Baerends RJ, Veenhuis M, van der Klei IJ.
FEMS Yeast Res. 2012 May;12(3):271-8. doi: 10.1111/j.1567-1364.2011.00772.x. Epub 2011 Dec 23. Review.
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