TY - BOOK

T1 - Protein Production Strategies for Gram-Positive Bacteria

T2 - An Approach for Cell Factories Optimization

AU - Ferro, Roberto

PY - 2017

Y1 - 2017

N2 - Humans have exploited microbes for centuries to produce cheese,beer and bread. Bacteria are not only part of the biosphere, but theyare also part of the flora in our guts and fundamental for life as weknow it. The huge potential of microbes is already harnessed todayby using their microbial cells as production facilities to produce fuels,chemicals, enzyme for detergents and pharmaceutical proteins suchas insulin.This study explores the potential of two bacteria commonly used infood fermentations, namely Lactococcus lactis and Bacillus subtilis, toproduce proteins for the biotech industry. A simple and inexpensiveapproach to create a growth based selection system that increasesprotein production is presented. The system can pick high proteinproducers out of a collection of protein producing variants, by usinga coupling device that ties expression of the gene of interest to an antibioticresistance gene. The coupling mechanism is further elucidatedin a patent application, which reports the development of multiplestrength levels for the coupling device and demonstrate its viabilityin Gram-negative bacteria (Escherichia coli and Pseudomonas putida).Together, the results reported in this PhD project expand theknowledge on Gram-positive cell factories and introduce a new toolthat will likely benefit the optimization of future ones. Coming studieswill build on the established platform and provide further insightin protein production

AB - Humans have exploited microbes for centuries to produce cheese,beer and bread. Bacteria are not only part of the biosphere, but theyare also part of the flora in our guts and fundamental for life as weknow it. The huge potential of microbes is already harnessed todayby using their microbial cells as production facilities to produce fuels,chemicals, enzyme for detergents and pharmaceutical proteins suchas insulin.This study explores the potential of two bacteria commonly used infood fermentations, namely Lactococcus lactis and Bacillus subtilis, toproduce proteins for the biotech industry. A simple and inexpensiveapproach to create a growth based selection system that increasesprotein production is presented. The system can pick high proteinproducers out of a collection of protein producing variants, by usinga coupling device that ties expression of the gene of interest to an antibioticresistance gene. The coupling mechanism is further elucidatedin a patent application, which reports the development of multiplestrength levels for the coupling device and demonstrate its viabilityin Gram-negative bacteria (Escherichia coli and Pseudomonas putida).Together, the results reported in this PhD project expand theknowledge on Gram-positive cell factories and introduce a new toolthat will likely benefit the optimization of future ones. Coming studieswill build on the established platform and provide further insightin protein production

UR - https://soeg.kb.dk/permalink/45KBDK_KGL/fbp0ps/alma99122738210005763

M3 - Ph.D. thesis

BT - Protein Production Strategies for Gram-Positive Bacteria

PB - Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen

ER -