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Full-Text Articles in Engineering
Rapid Characterization And Engineering Of Natural Product Biosynthetic Pathways Via Dna Assembler, Zengyi Shao, Yunzi Luo, Huimin Zhao
Rapid Characterization And Engineering Of Natural Product Biosynthetic Pathways Via Dna Assembler, Zengyi Shao, Yunzi Luo, Huimin Zhao
Zengyi Shao
We report a synthetic biology strategy for rapid genetic manipulation of natural product biosynthetic pathways. Based on DNA assembler, this method synthesizes the entire expression vector containing the target biosynthetic pathway and the genetic elements required for DNA maintenance and replication in various hosts in a single-step manner through yeast homologous recombination, offering unprecedented flexibility and versatility in pathway manipulations.
Cloning And Heterologous Expression Of The Spectinabilin Biosynthetic Gene Cluster From Streptomyces Spectabilis, Zengyi Shao
Cloning And Heterologous Expression Of The Spectinabilin Biosynthetic Gene Cluster From Streptomyces Spectabilis, Zengyi Shao
Zengyi Shao
Spectinabilin is a rare nitrophenyl-substituted polyketide metabolite. Here we report the cloning and heterologous expression of the spectinabilin gene cluster from Streptomyces spectabilis. Unexpectedly, this gene cluster is evolutionarily closer to the aureothin gene cluster than to the spectinabilin gene cluster from Streptomyces orinoci. Moreover, the two nearly identical spectinabilin gene clusters use a distinctly different regulation mechanism.
Characterizing Volatile Fatty Acids And Other Gases In A Rumen Closed In Vitro Fermentation System Using Solid Phase Microextraction, Jarett P. Spinhirne, Jacek A. Koziel, Norbert K. Chirase
Characterizing Volatile Fatty Acids And Other Gases In A Rumen Closed In Vitro Fermentation System Using Solid Phase Microextraction, Jarett P. Spinhirne, Jacek A. Koziel, Norbert K. Chirase
Jacek A. Koziel
A new method for rapid sampling and qualitative characterization of the headspace gases of closed in vitro cultures using solid phase microextraction (SPME) was evaluated for ruminal fluid and ruminal fluid with feed containing a feed additive. Gas sample collection was achieved by exposing a DVB/Carboxen/PDMS 50/30 .m SPME fiber to the headspace of cultures for 1 min every hour. This was followed by immediate analysis on a GC–MS and then reuse of the SPME fiber. Acetic, propionic, isobutyric, butyric, isovaleric, valeric, and hexanoic acids and toluene, dimethyl disulfide, and pentadecane were identified in the headspace. SPME technology facilitated rapid …
Characterization Of Oxidation Products Of Tnt Metabolism In Aquatic Phytoremediation Systems Of Myriophyllum Aquaticum, R. Bhadra, R. J. Spanggord, D. G. Wayment, J. B. Hughes, Jacqueline V. Shanks
Characterization Of Oxidation Products Of Tnt Metabolism In Aquatic Phytoremediation Systems Of Myriophyllum Aquaticum, R. Bhadra, R. J. Spanggord, D. G. Wayment, J. B. Hughes, Jacqueline V. Shanks
Jacqueline V. Shanks
TNT transformation processes in sediment-free, “natural”, aquatic phytoremediation systems of Myriophyllum aquaticum were investigated with specific interest in oxidation products. Extraction procedures combining liquidliquid extractions and solid-phase extractions were developed for the isolation of the mostly acidic, oxidized TNT metabolites. Six compounds unique from the reduction products of TNT were isolated and characterized by UVvis, 1H, and 13C NMR spectroscopy, by mass spectroscopy, and by chemical synthesis where feasible. These compounds include 2-amino-4,6-dinitrobenzoic acid, 2,4-dinitro-6- hydroxy-benzyl alcohol, 2-N-acetoxyamino-4,6-dinitrobenzaldehyde, 2,4-dinitro-6-hydroxytoluene, and two binuclear metabolites unique from the customary azoxytetranitrotoluenes. The monoaryl compounds show clear evidence of oxidative transformations, methyl oxidation and/or …
Confirmation Of Conjugation Processes During Tnt Metabolism By Axenic Plant Roots, R. Bhadra, D. G. Wayment, J. B. Hughes, Jacqueline V. Shanks
Confirmation Of Conjugation Processes During Tnt Metabolism By Axenic Plant Roots, R. Bhadra, D. G. Wayment, J. B. Hughes, Jacqueline V. Shanks
Jacqueline V. Shanks
This paper examines processes in plants for the formation of fate products of TNT beyond its aminated reduction products, 2-amino-4,6-dinitrotoluene and 4-amino-2,6- dinitrotoluene. TNT metabolites were isolated and characterized in combination with temporal analyses of product profiles and 14C distribution, in microbe-free, axenic root cultures of Catharanthus roseus. Four unique TNTderived compounds were isolated. Using evidence from 1H NMR, mass spectroscopy, HPLC, acid hydrolysis, and enzymatic hydrolysis with â-glucuronidase and â-glucosidase, they were established as conjugates formed by reactions of the amine groups of 2-amino-4,6-dinitrotoluene and 4-amino-2,6-dinitrotoluene. From the mass spectral evidence, at least a six-carbon unit from the plant …