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Biochemical and Biomolecular Engineering Commons™
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Articles 1 - 4 of 4
Full-Text Articles in Biochemical and Biomolecular Engineering
Quantifying And Resolving Multiple Vector Transformants In S. Cerevisiae Plasmid Libraries, Thomas C. Scanlon, Elizabeth C. Gray, Karl E. Griswold
Quantifying And Resolving Multiple Vector Transformants In S. Cerevisiae Plasmid Libraries, Thomas C. Scanlon, Elizabeth C. Gray, Karl E. Griswold
Dartmouth Scholarship
In addition to providing the molecular machinery for transcription and translation, recombinant microbial expression hosts maintain the critical genotype-phenotype link that is essential for high throughput screening and recovery of proteins encoded by plasmid libraries. It is known that Escherichia coli cells can be simultaneously transformed with multiple unique plasmids and thusly complicate recombinant library screening experiments. As a result of their potential to yield misleading results, bacterial multiple vector transformants have been thoroughly characterized in previous model studies. In contrast to bacterial systems, there is little quantitative information available regarding multiple vector transformants in yeast. Saccharomyces cerevisiae is the …
Enzyme–Microbe Synergy During Cellulose Hydrolysis By Clostridium Thermocellum, Yanpin Lu, Yi-Heng P. Zhang, Lee R. Lynd
Enzyme–Microbe Synergy During Cellulose Hydrolysis By Clostridium Thermocellum, Yanpin Lu, Yi-Heng P. Zhang, Lee R. Lynd
Dartmouth Scholarship
Specific cellulose hydrolysis rates (g of cellulose/g of cellulase per h) were shown to be substantially higher (2.7- to 4.7-fold) for growing cultures of Clostridium thermocellum as compared with purified cellulase preparations from this organism in controlled experiments involving both batch and continuous cultures. This “enzyme–microbe synergy” requires the presence of metabolically active cellulolytic microbes, is not explained by removal of hydrolysis products from the bulk fermentation broth, and appears due to surface phenomena involving adherent cellulolytic microorganisms. Results support the desirability of biotechnological processes featuring microbial conversion of cellulosic biomass to ethanol (or other products) in the absence of …
Use Of Combinatorial Genetic Libraries To Humanize N-Linked Glycosylation In The Yeast Pichia Pastoris, Byung-Kwon Choi, Piotr Bobrowicz, Robert C. Davidson, Stephen R. Hamilton, David Kung, Huijuan Li, Robert Miele, Juergen Nett, Stefan Wildt, Tillman Gerngross
Use Of Combinatorial Genetic Libraries To Humanize N-Linked Glycosylation In The Yeast Pichia Pastoris, Byung-Kwon Choi, Piotr Bobrowicz, Robert C. Davidson, Stephen R. Hamilton, David Kung, Huijuan Li, Robert Miele, Juergen Nett, Stefan Wildt, Tillman Gerngross
Dartmouth Scholarship
The secretory pathway of Pichia pastoris was genetically re-engineered to perform sequential glycosylation reactions that mimic early processing of N-glycans in humans and other higher mammals. After eliminating nonhuman glycosylation by deleting the initiating alpha-1,6-mannosyltransferase gene from P. pastoris, several combinatorial genetic libraries were constructed to localize active alpha-1,2-mannosidase and human beta-1,2-N-acetylglucosaminyltransferase I (GnTI) in the secretory pathway. First, >32 N-terminal leader sequences of fungal type II membrane proteins were cloned to generate a leader library. Two additional libraries encoding catalytic domains of alpha-1,2-mannosidases and GnTI from mammals, insects, amphibians, worms, and fungi were cloned to generate catalytic domain libraries. …
Microbial Cellulose Utilization: Fundamentals And Biotechnology, Lee R. Lynd, Paul J. Weimer, Willem H. Van Zyl, Isak S. Pretorius
Microbial Cellulose Utilization: Fundamentals And Biotechnology, Lee R. Lynd, Paul J. Weimer, Willem H. Van Zyl, Isak S. Pretorius
Dartmouth Scholarship
Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial …