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Full-Text Articles in Biomedical Engineering and Bioengineering

Dna Assembler, An In Vivo Genetic Method For Rapid Construction Of Biochemical Pathways, Zengyi Shao, Hua Zhao, Huimin Zhao Jan 2009

Dna Assembler, An In Vivo Genetic Method For Rapid Construction Of Biochemical Pathways, Zengyi Shao, Hua Zhao, Huimin Zhao

Zengyi Shao

The assembly of large recombinant DNA encoding a whole biochemical pathway or genome represents a significant challenge. Here, we report a new method, DNA assembler, which allows the assembly of an entire biochemical pathway in a single step via in vivo homologous recombination in Saccharomyces cerevisiae. We show that DNA assembler can rapidly assemble a functional d-xylose utilization pathway (∼9 kb DNA consisting of three genes), a functional zeaxanthin biosynthesis pathway (∼11 kb DNA consisting of five genes) and a functional combined d-xylose utilization and zeaxanthin biosynthesis pathway (∼19 kb consisting of eight genes) with high efficiencies (70 - 100%) …


Rational Pathway Engineering Of Type I Fatty Acid Synthase Allows The Biosynthesis Of Triacetic Acid Lactone From D-Glucose In Vivo, Wenjuan Zha, Zengyi Shao, John W. Frost, Huimin Zhao Jan 2004

Rational Pathway Engineering Of Type I Fatty Acid Synthase Allows The Biosynthesis Of Triacetic Acid Lactone From D-Glucose In Vivo, Wenjuan Zha, Zengyi Shao, John W. Frost, Huimin Zhao

Zengyi Shao

Metabolic pathway engineering is a powerful tool to synthesize structurally diverse and complex chemicals via genetic manipulation of multistep catalytic systems involved in cell metabolism. Here, we report the rational design of a fatty acid biosynthetic pathway, Brevibacterium ammoniagenes fatty acid synthase B (FAS-B), that allows the microbial synthesis of triacetic acid lactone (TAL) from an inexpensive feedstock, D-glucose. TAL can be chemically converted to phloroglucinol, which is a core structure for the synthesis of various high value bioactive compounds and energetic compounds such as 1,3,5-triamino-2,4,6-trinitrobenzene (TATB). Synthesis of phloroglucinol from d-glucose using this combined biological and chemical synthesis may …