- 期刊
Sequence Analysis for Metabolic Evolution of Archaeoglobus fulgidus Geranylgeranyl Diphosphate Synthase in Escherichia coli for Carotenoid Biosynthesis
並列摘要
To investigate possible ways for improving the metabolic output after horizontal gene transfer, we mutated a DNA fragment from the hyperthermophile Archaeoglobus fulgidus containing geranylgeranyl pyrophosphate synthase (GPS), and screened for higher metabolic output in an Escherichia coli strain that expressed the carotenoid biosynthesis gene cluster (crtBIYZW). A total of 16 independent, positive mutants were isolated, sequenced, and analyzed. A binomial statistical analysis identified a high mutation frequency region upstream of the protein coding region, which may be responsible for optimizing gene expression level in the new host. The same analysis also identified two low frequency regions within the protein coding region. These two regions represent the highly conserved regions during evolution and contain the presumed active site domains. Mutations in the protein coding region also showed a trend of amino acid substitutions that increase structural flexibility, a reversal of trend for increasing thermostability. This work provides a rapid and practical method to map important regions for function, and obtained mutants that are useful for biosynthesis of carotenoids.
延伸閱讀
- 劉逸琪、張淑貞、陳文華、許永斌（2006）。The application of single-step nested multiplex polymerase chain reaction for the identification of Rhizoglyphus echinopus, R. robini and R. setosus simultaneously。植物保護學會會刊，48(2)，101-116。https://doi.org/10.6715/PPB.200606_48(2).0004
- 許怡亭（2006）。Analysis of exopolysaccharide production and key enzyme activities of several lactobacilli and bifidobacteria〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU.2006.00945
- Kudva, I. T., Davis, M. A., Griffin, R. W., Garren, J., Murray, M., John, M., Hovde, C. J., & Calderwood, S. B. (2012). Polymorphic Amplified Typing Sequences and Pulsed-Field Gel Electrophoresis Yield Comparable Results in the Strain Typing of a Diverse Set of Bovine Escherichia coli O157:H7 Isolates. International Journal of Microbiology, 2012(), 17-26. https://doi.org/10.1155/2012/140105
- Tetteh, A. Y., Sun, K. H., Hung, C. Y., Kittur, F. S., Ibeanu, G. C., Williams, D., & Xie, J. H. (2014). Transcriptional Response of Selenopolypeptide Genes and Selenocysteine Biosynthesis Machinery Genes in Escherichia coli during Selenite Reduction. International Journal of Microbiology, (2014), 126-136. https://doi.org/10.1155/2014/394835
- 鄭宇辰（2021）。Conversion of Escherichia coli into mixotrophic CO2 assimilation with malate and hydrogen based on recombinant expression of 2-oxoglutarate:ferredoxin oxidoreductase using adaptive laboratory evolution〔碩士論文，國立清華大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0016-0203202211552401