BioChem - DOS

Identification and use of an alkane transporter plug-in for applications in biocatalysis and whole-cell biosensing of alkanes

Scientific reports

Grant, Chris; Deszcz, Dawid; Wei, Yu-Chia; Martí nez-Torres, Rubé ns Julio; Morris, Phattaraporn; Folliard, Thomas; Sreenivasan, Rakesh; Ward, John; Dalby, Paul; Woodley, John M.; Baganz, Frank

<P>Effective application of whole-cell devices in synthetic biology and biocatalysis will always require consideration of the uptake of molecules of interest into the cell. Here we demonstrate that the AlkL protein from <I>Pseudomonas putida</I> GPo1 is an alkane import protein capable of industrially relevant rates of uptake of C<SUB>7</SUB>-C<SUB>16</SUB> n-alkanes. Without alkL expression, native <I>E.coli</I> n-alkane uptake was the rate-limiting step in both the whole-cell bioconversion of C<SUB>7</SUB>-C<SUB>16</SUB> n-alkanes and in the activation of a whole-cell alkane biosensor by C<SUB>10</SUB> and C<SUB>11</SUB> alkanes. By coexpression of alkL as a transporter plug-in, specific yields improved by up to 100-fold for bioxidation of >C<SUB>12</SUB> alkanes to fatty alcohols and acids. The alkL protein was shown to be toxic to the host when overexpressed but when expressed from a vector capable of controlled induction, yields of alkane oxidation were improved a further 10-fold (8&#x2005;g/L and 1.7&#x2005;g/g of total oxidized products). Further testing of activity on n-octane with the controlled expression vector revealed the highest reported rates of 120&#x2005;&mu;mol/min/g and 1&#x2005;g/L/h total oxidized products. This is the first time AlkL has been shown to directly facilitate enhanced uptake of C<SUB>10</SUB>-C<SUB>16</SUB> alkanes and represents the highest reported gain in product yields resulting from its use.</P>

2014

Nature Publishing Group

cc-by

2045-2322

4

pp.5844

10.1038/srep05844

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART72142331

https://www.nature.com/articles/srep05844.pdf