Identification, isolation, and heterologous expression of Sunflower wax synthase for the synthesis of tailored wax esters

2020 ◽  
Vol 44 (10) ◽  
Author(s):  
Theresa Shalini ◽  
Asha Martin
Keyword(s):  
Heterologous Expression ◽  
Wax Esters ◽  
Wax Synthase ◽  
Sunflower Wax

scholarly journals Detailed characterization of the substrate specificity of mouse wax synthase.

2013 ◽  
Vol 60 (2) ◽  
Author(s):  
Magdalena Miklaszewska ◽  
Adam Kawiński ◽  
Antoni Banaś
Keyword(s):  
Substrate Specificity ◽  
Large Scale ◽  
Fatty Acyl ◽  
Wax Esters ◽  
In Vitro Assays ◽  
Esterification Reaction ◽  
Scale Production ◽  
Acyltransferase Activity ◽  
Wax Synthase

Wax synthases are membrane-associated enzymes catalysing the esterification reaction between fatty acyl-CoA and a long chain fatty alcohol. In living organisms, wax esters function as storage materials or provide protection against harmful environmental influences. In industry, they are used as ingredients for the production of lubricants, pharmaceuticals, and cosmetics. Currently the biological sources of wax esters are limited to jojoba oil. In order to establish a large-scale production of desired wax esters in transgenic high-yielding oilseed plants, enzymes involved in wax esters synthesis from different biological resources should be characterized in detail taking into consideration their substrate specificity. Therefore, this study aims at determining the substrate specificity of one of such enzymes -- the mouse wax synthase. The gene encoding this enzyme was expressed heterologously in Saccharomyces cerevisiae. In the in vitro assays (using microsomal fraction from transgenic yeast), we evaluated the preferences of mouse wax synthase towards a set of combinations of 11 acyl-CoAs with 17 fatty alcohols. The highest activity was observed for 14:0-CoA, 12:0-CoA, and 16:0-CoA in combination with medium chain alcohols (up to 5.2, 3.4, and 3.3 nmol wax esters/min/mg microsomal protein, respectively). Unsaturated alcohols longer than 18°C were better utilized by the enzyme in comparison to the saturated ones. Combinations of all tested alcohols with 20:0-CoA, 22:1-CoA, or Ric-CoA were poorly utilized by the enzyme, and conjugated acyl-CoAs were not utilized at all. Apart from the wax synthase activity, mouse wax synthase also exhibited a very low acyl-CoA:diacylglycerol acyltransferase activity. However, it displayed neither acyl-CoA:monoacylglycerol acyltransferase, nor acyl-CoA:sterol acyltransferase activity.

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