Enzyme Mixtures and Complex Biosynthesis (Biotechnology by Sanjoy K. Bhattacharya, M. Tech., Ph.D.

By Sanjoy K. Bhattacharya, M. Tech., Ph.D.

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Production of inulooligosaccharides from inulin by a dual endoinulinase system. Enz Microb Technol 2001; 29:428-433. 32. Olano-Mamrtin E, Mountzouris KC, Gibson GR et al. Continuous production of pectic oligosaccharides in an enzyme membrane reactor.

This process mimics the natural recombination that occurs during sexual reproduction. The great advantage of directed evolution techniques is that they require no prior knowledge of the structure of the enzyme, nor it is necessary to be able to predict what effect a certain mutation will have. The results of these techniques are often surprising in that desired changes are often produced by unexpected mutations. The disadvantage is that they require high-throughput, which is not always feasible.

Still thousands (of ~10,000) of them, remain to be identified in terms of their three-dimensional structure. Researchers with limited resources perform random mutagenesis on a selected gene followed by expression and testing. When little is known about the structure of an enzyme, numerous mutagenized enzymes are generated rapidly and screened for changes in their characteristics. Mutants with desired characteristics are then sequenced to identify the region that determines the characteristic. The method mimics natural evolution and although a random process, it may produce superior results 20 Enzyme Mixtures and Complex Biosynthesis Table 1.

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