A broad‐range yeast expression system reveals Arxula adeninivorans expressing a fungal self‐sufficient cytochrome P450 monooxygenase as an excellent whole‐cell biocatalyst. Issue 4 (5th March 2014)

• Record Type: Journal Article
• Authors: Theron, Chrispian W.; Labuschagné, Michel; Gudiminchi, Ramakrishna; Albertyn, Jacobus; Smit, Martha S.
• Is Part Of: FEMS yeast research. Volume 14:Issue 4(2014)
• Page Start: 556
• Page End: 566
• Publication Date: 2014-03-05
• Subjects: Yeast -- Periodicals; Yeasts -- Periodicals; 579.562
• DOI: 10.1111/1567-1364.12142
• Languages: English
• ISSNs: 1567-1356

Abstract

<abstract abstract-type="main" id="fyr12142-abs-0001"> <title>Abstract</title> <p>The feasibility of using a single vector to clone a cytochrome P450 monooxygenase (P450) in different yeasts and then compare whole‐cell hydroxylase activity was investigated. A broad‐range yeast expression vector using the ylTEFp to drive expression of the cloned gene and the scTEFp to drive the hygromycin resistance marker gene was used to clone the genes encoding two self‐sufficient P450s, CYP102A1 and CYP505A1. Both genes were cloned into <italic>Saccharomyces cerevisiae</italic>, <italic> Kluyveromyces marxianus</italic>, <italic> Yarrowia lipolytica</italic> (two strains) and <italic>Arxula adeninivorans</italic>. 4‐Hexylbenzoic acid (HBA), which is subterminally hydroxylated by both CYP102A1 and CYP505A1, was used to compare whole‐cell hydroxylase activity of transformants. <italic>Kluyveromyces marxianus</italic> and <italic>A. adeninivorans</italic> exhibited activity with both CYP102A1 and CYP505A1, while <italic>S. cerevisiae</italic> only displayed CYP102A1 activity and <italic>Y. lipolytica</italic> only CYP505A1 activity. The highest CYP102A1 activity (0.8 mM HBA converted in 24 h) was observed with concentrated resting‐cell suspensions of <italic>S. cerevisiae</italic>. The CYP505A1 activity observed with growing cultures of <italic>A. adeninivorans</italic> was however at least 12 times higher than the CYP102A1 activity of <italic>S. cerevisiae</italic> with up to 2 mM HBA converted within 6 h. The use of <italic>K. marxianus</italic> and <italic>A. adeninivorans</italic> for P450 expression has not previously been reported.</p> </abstract>