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Dr. Jung's MIT Research


New functional protein chip for in vitro metabolic engineering:

                         Functional protein chips are critical for the next phase of proteomics research. Like DNA chips, protein chips will be able to analyze thousands of samples simultaneously, leading the way towards a complete map of the entire complement of human proteins. But, unlike DNA, proteins are not so easy to attach to chips. Where DNA is robust, and able to withstand harsh experimental conditions, proteins are fragile and will denature if they aren't treated gently. While there are good methods for amplifying DNA, so that even very tiny amounts can be detected, there are none available for proteins. It's a whole new set of constraints. To accommodate large number of proteins and minimize preparation time using genomic information only, a simple method avoiding long and expensive protein purification steps is required. In this research, a new functional protein chip employing RNA display technology is investigated. This method uses specific hybridization of RNA-protein fusion molecules, linked covalently by a puromycin-oligoDNA linker, on oligo-spotted cDNA microarrays.


Metabolic engineering of lysine production pathway in Corynebacterium glutamicum:

                         L-Lysine production by C. glutamicum has been emphasized as a major target of metabolic engineering because of the commercial importance of lysine and the appropriate metabolic complexity of lysine production pathway interrelated with cell growth, energy production and product synthesis. Metabolic engineering generally consists of two parts: (a) application of biochemical and mathematical analyses in order to identify target metabolic pathways and assess cell physiology and (b) pathway modification by using molecular biological techniques including gene cloning, site-directed mutagenesis, etc. As a result of the accumulated studies, it was found that pyruvate carboxylase (PC), responsible for the anaplerotic reaction from pyruvate (PYR) to oxaloacetate (OAA), plays a key role in amino acid production improvement. As a subsequent study, pc gene coding for PC was cloned and overexpressed in C. glutamicum.

                         Recently, coordinated overexpression of pc, responsible for anaplerosis, combined with ask in C. glutamicum was conducted and its physiological features were investigated. Compared to the wild type strain, the new recombinant strain showed higher lysine productivity than the wild type without any significant change of growth. From these results, we proceed that coordinated overexpression of pc coupled with other limiting downstream pathways to improve amino acid productivity of C. glutamicum. The other candidates in the lysine synthesis pathway, e.g., DDP synthase and permease should be amplified also together with pc and ask genes.


Publications

  • Gyoo Yeol Jung and Gregory Stephanopoulos; (2004): A Functional Protein Chip for Pathway Optimization and in Vitro Metabolic Engineering Science Volume 304, Number 5669; 428-431 (16 April 2004).

  • Ugarov, V. I., Morozov, I. Yu, Jung, G. Y., Chetverin, A. B., and Spirin, A. S.; (1994): Expression and stability of RQ-messenger RNA in cell-free translation systems. - FEBS Letters, 341, 131-134.

  • Jung, G. Y., Lee, E. Y., Kim, Y.-E., Jung, B. W., Kang, S.-H., and Choi, C. Y.; (2000): Stabilization effect of zeolite on DHFR mRNA in wheat germ cell-free protein synthesis system. - J. Biosci. Bioeng., 89, 193-195.

  • Jung, G. Y., Jung, H. O., Kim, J. R., Ahn, Y., and Park, S.; (1999): Isolation and characterization of Rhodopseudomonas palustris P4 which utilizes CO with the production of H2. - Biotechnol. Letters, 21, 525-529.

  • Jung, G. Y., Kim, J. R., Jung, H. O., Park, J.-Y., and Park, S.; (1999): A new chemoheterotrophic bacterium catalyzing water-gas shift reaction. - Biotechnol. Lett., 21, 869-873.

  • Jung, G. Y. and Choi, C. Y.; (2001): Improvement of specific protein synthesis activity by using partially fortified wheat germ cell-free system. - Biotechnol. Lett.; Submitted.

  • Jung, G. Y., Kim, J. R., Jung, H. O., Park, J.-Y., and Park, S.; (2001): Carbon-monoxide oxidizing falcultatively anaerobic Citrobacter Sp. - J. Microbiol.Biotechnol.; Submitted.

  • Jung, G. Y., Kim, J. R., Jung, H. O., Park, J.-Y., and Park, S.; (2001): Hydrogen production by Citrobacter Sp. Y19. - Int. J. Hydr. Energy, 27, 601-610.

  • Koffas, M. A. G., Jung, G. Y., Aon, J. C., and Stephanopoulos, G.; (2001): Effect of pyruvate carboxylase overexpression on physiology of Corynebacterium glutamicum. - Appl. Environ. Microbiol.; accepted.

  • Koffas, M. A. G., Jung, G. Y., and Stephanopoulos, G.; (2001): Coordinated overexpression of pc and ask for development of metabolic engineered Corynebacterium glutamicum. - Metabolic Engineering; Submitted.

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