Engineering of Penicillium chrysogenum

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• 作者：Diana M. Harris ; Ilja Westerlaken ; Dick Schipper ; Zita A. van der Krogt ; Andreas K. Gombert ; John Sutherl ; Leonie M. Raamsdonk ; Marco A. van den Berg ; Roel A.L. Bovenberg ; Jack T. Pronk ; Jean-Marc Da
• 关键词：Penicillium chrysogenum ; Metabolic engineering ; Adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid ; Carbamoylated cephem antibiotics
• 刊名：Metabolic Engineering
• 出版年：2009
• 出版时间：March 2009
• 年：2009
• 卷：11
• 期：2
• 页码：125-137
• 全文大小：1118 K

文摘

Penicillium chrysogenum was successfully engineered to produce a novel carbamoylated cephalosporin that can be used as a synthon for semi-synthetic cephalosporins. To this end, genes for Acremonium chrysogenum expandase/hydroxylase and Streptomyces clavuligerus carbamoyltransferase were expressed in a penicillinG high-producing strain of P. chrysogenum. Growth of the engineered strain in the presence of adipic acid resulted in production of adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid (ad7-ACCCA) and of several adipoylated pathway intermediates. A combinatorial chemostat-based transcriptome study, in which the ad7-ACCCA-producing strain and a strain lacking key genes in β-lactam synthesis were grown in the presence and absence of adipic acid, enabled the dissection of transcriptional responses to adipic acid per se and to ad7-ACCCA production. Transcriptome analysis revealed that adipate catabolism in P. chrysogenum occurs via β-oxidation and enabled the identification of putative genes for enzymes involved in mitochondrial and peroxisomal β-oxidation pathways. Several of the genes that showed a specifically altered transcript level in ad7-ACCCA-producing cultures were previously implicated in oxidative stress responses.