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Fig. 1 | Biotechnology for Biofuels

Fig. 1

From: Rapid optimisation of cellulolytic enzymes ratios in Saccharomyces cerevisiae using in vitro SCRaMbLE

Fig. 1

Schematic of the in vitro SCRaMbLE strategy to optimise the gene copy number and ratio of CEL3A and CEL5A for efficient synergistic enzyme activity. A DNA library was generated by SCRaMbLEing donor DNA (loxPsym-flanked sequences in pCEL3A-loxP and pCEL5A-loxP) in acceptor plasmids (pAcceptor) (a). The resulting library (b) contained plasmids with various copy numbers of CEL3A and CEL5A for direct transformation into S. cerevisiae (c) which conferred a range of synergistic cellulase activity, determined utilising the BPNPG5 substrate (d). The copy number of CEL3A and CEL5A genes which enabled the highest enzyme activity was determined by quantitative PCR (e)

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Biotechnology for Biofuels and Bioproducts

ISSN: 2731-3654

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