Although the routes of de novo pyridoxal 5'-phosphate (PLP) biosynthesis have been well described, studies of the engineering of an intracellular PLP supply are limited, and the effects of cellular PLP levels on PLP-dependent enzyme-based whole-cell biocatalyst activity have not been described. To investigate the effects of PLP cofactor availability on whole-cell biocatalysis, the ribose 5-phos...

Background: Novozym 435, a commercial lipase from Candida antarctica, recombinant, expressed in Aspergillus niger, immobilized on macroporous acrylic resin, has been already described in the obtention of biodiesel. It is here evaluated in the production of a new biofuel that integrates the glycerol as monoglyceride (MG) together with two fatty acid ethyl esters (FAEE) molecules by the applicati...

In order to operate a process which uses immobilized enzymes at constant conversion and constant capacity, the refreshment of the enzyme must be continuous. In this paper, two reactor types with continuous refreshment of the biocatalyst are discussed: the stirred tank and the multistage fluidized bed. A method is presented for dimensioning a reactor in such a way that the costs for the conversi...

The styrene oxide isomerase (SOI) represents a membrane-bound enzyme of the microbial styrene degradation pathway and has been discussed as promising biocatalyst. It catalyzes the isomerization of styrene oxide to phenylacetaldehyde. In this study a styC gene, which encodes the SOI of Rhodococcus opacus 1CP, was optimized for optimal expression in Escherichia coli BL21(DE3) pLysS. The expressio...

Considered one of the most important enzymes for the biotechnology industries, lipase is a very versatile biocatalyst been target of several researches and review papers and used in many different industrial applications, such as household detergent additive, chemicals and pharmaceutical compounds synthesis, biodiesel production and others. Lipase produced by Burkholderia cepacia was entrapped ...

Multienzyme processes represent an important area of biocatalysis. Their efficiency can be enhanced by optimization of the stoichiometry of the biocatalysts. Here we present a workflow for maximizing the efficiency of a three-enzyme system catalyzing a five-step chemical conversion. Kinetic models of pathways with wild-type or engineered enzymes were built, and the enzyme stoichiometry of each ...