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It could be envisaged that performing saturation mutagenesis at these positions would yield a more thermostable variant, an approach successfully employed by Reetz et al. (2006).
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As no Udh is known from thermophilic organisms, we developed more thermostable variants by enzyme engineering.
To overcome this limitation, we created more thermostable variants using semi-rational design.
Directed evolution of cold-adapted enzymes to a more thermostable version, appears as an attractive strategy to fulfill the stability and activity requirements for the industry.
A more thermostable version of an enzyme is highly desirable as it would increase the productivity of enzyme conversion [ 24].
It is clear from our study, however, that care should be exercised when using B-factors alone to design more thermostable protein variants.
Although all nine mutants were overexpressed, soluble, contained covalently bound flavin and were active with xylitol, unfolding experiments revealed that none of the AldO mutants was more thermostable than the wild-type variant.
In comparison to native protein LipJ, point variant Ala81Tyr was observed to be more thermostable and demonstrated 1.6 times relative activity after 1 h exposure at 50 °C.
The cellulase of strain RG-07 is more thermostable than cellulase studied by several other researchers.
Hence it is evident that the cellulase of strain RG-07 is more thermostable, and may be applied to several biotechnological and industrial purposes.
For example, a thermostable variant expressed in biofuel feedstocks could be envisaged for removing hemicellulose more efficiently during pretreatment and for reducing enzyme loads during enzymatic hydrolysis.
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