Solubility proteins, 7252 review

A major feature of HTP protein production pipelines is the inclusion of a screening step at a relatively small scale to identify constructs suitable in terms of soluble protein yield for scale-up and subsequent protein purification. Given the relatively high cost of the latter steps in terms of time and resources, particularly if eukaryotic expression is required, the screening stage is seen as crucial to the overall process. In this section the approaches to the evaluation of expression at small scale will be reviewed. 

Methanol dehydrogenase (MDH EC 1.1.99.8) catalyses the oxidation of methanol to formaldehyde in the periplasm of methylotrophic baeteria during growth on methanol or methane. It was first deseribed in Methy-lobacterium extorquens (Anthony and Zatman, 1964a,b) and has subsequently been shown to be the one feature that is eommon to almost all methylotrophs in which it often constitutes up to 15% of their soluble protein (see Anthony, 1986 for a review of the basie enzymology of a wide range of MDHs). 

Despite the limitations, the great success of enzyme immobilization in diagnostics, pharmaceutical, food and chemical industries is undeniable12, 3. The decision whether one should use a soluble enzyme preparation or an immobilized enzyme does not have a universal solution and can be decided only on a case by case basis. Ordinarily, if the cost of an enzyme represents a significant portion of the overall cost or if isolation of the final product is complicated by the presence of the soluble protein, the cost of immobilization can be offset by the gains in productivity and improved product quality. The intent of this section is to describe, in general terms with illustrative examples, the features and considerations of these broad classes of enzyme immobilization as they impact their application to biocatalysis. Detailed experimental protocols are available in the original literature and exemplary protocols for these methods are offered in many excellent reviews and texts 14L... 

While several lines of evidence show that jasmonic acid and/or methyl jasmonate are endogenous signal transducers in the tendril s response to mechano-stimulation [79-81], evidence has been obtained for the constitutive expression of LOX in the tendril of B. dioica [82]. Tendril LOX is apparently a soluble protein with an alkaline pH optimum ( 8.5) and its major product is 13-HpOT. Thus, the tendril LOX has characteristics typical of class-1 enzymes (for review, see [28]), where 13-hydroperoxide is formed from 18 3. 

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