Denaturation efficiency ratio

In PCR product, the untreated sample can be considered as a strand-specific intra-assay negative control. Electrochemical signals upon thermal denaturation undoubtedly designated the presence of a complementary sequence. Genosensors showed a different behaviour as underlined by the signal ratio between the denaturated and non-denaturated sample that highlighted the dissimilar efficiency of each probe. 

Sulfonated surfactants are also powerful protease deactivators [67]. The presence of the sulfonate group on benzene makes sodium dodecyl benzene sulfonate, also called LAS (linear alkylbenzene sulfonate), more surface active than SDS. As a result, LAS is an even more efficient denaturant than SDS [27] and alcohol ethoxy sulfates [32]. For instance, in a 0.02% LAS solution, the activity of the protease from Bacillus stearothermophilus is reduced to 20% of its value in water [68]. Likewise, the subtilisin saturation by surfactants at pH 7.4 has been reported to occur at surfactant to enzyme ratios (w/w) of 6.3 (SDS) and 3.2 (LAS) [69]. 

Other parameters of interest in the enzymatic processing of bioactive compounds include reaction temperatnre, agitation level and the selection of substrate types and ratio. Firstly, temperatnre can inflnence not only the activation and denaturation of the enzyme, but also substrate and prodnct solnbility and the viscosity of the reaction media (Chebil et al, 2006 Villenenve, 2007). Particnlarly when viscons snbstrates or ionic liquids are involved, a well mixed system is very important to ensnre proper contact between the biocatalyst and snbstrates. From the preceding brief discnssion, it is clear that the efficiency of enzymatic processing of bioactive componnds is affected by anumber of parameters. Excellent in-depth discnssions of some key parameters can be fonnd in various reviews (Lau et al, 2004 Chebil et al., 2006 Villeneuve, 2007). 

To achieve high covalent-immobilization efficiency, several important factors must be optimized including protein load, pH, ionic strength, temperature, mass volume ratio, and agitation rate [70,71]. Moreover, it is sometimes necessary to add protective molecules such as carbohydrates and polyethyleneglycol to avoid enzyme denaturation and/or inactivation during conjugations. 

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