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首页> 外文学位 >Genetic analysis of phenylalanine and tyrosine degradation by Burkholderia cepacia DBO1.
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Genetic analysis of phenylalanine and tyrosine degradation by Burkholderia cepacia DBO1.

机译：洋葱伯克霍尔德菌DBO1降解苯丙氨酸和酪氨酸的遗传分析。

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摘要

The metabolism of the two aromatic amino acids phenylalanine and tyrosine are related in most bacterial species. Phenylalanine and tyrosine are synthesized from chorismate through phenylpyruvate and p-hydroxyphenylpyruvate respectively. The canonical pathway for the catabolism of phenylalanine proceeds through initial hydroxylation of phenylalanine to tyrosine. However, the catabolism of phenylalanine is not well understood in some microorganisms and recently one alternate pathway for phenylalanine degradation involving phenylacetate as an intermediate was described. Burkholderia cepacia DBO1 utilizes separate catabolic pathways for the utilization of phenylalanine and tyrosine. In order to clarify phenylalanine degradation in B. cepacia DBO1 we made extensive use of plasposon-based random insertional mutagenesis to generate mutants in the catabolic pathways of interest. Four mutant strains were obtained that could not grow on tyrosine and ten strains were obtained that that could not grow on phenylalanine. Interestingly, all the tyrosine catabolism mutant strains grew on phenylalanine, indicating that phenylalanine is not degraded through tyrosine. Sequence analysis of the phenylalanine mutant strains showed that the transposon insertions fall into three groups: an operon encoding phenylacetate degradation, a gene encoding a regulator, and a gene encoding pyruvate oxidoreductase. These data, combined with HPLC metabolite accumulation studies indicate that degradation of phenylalanine proceeds through phenylacetate. The enzymatic function of the proteins predicted to be involved in the catabolic pathway were examined through suitable biochemical techniques (enzymes assays and metabolite identification). RT-PCR experiments were used to examine the regulation of the genes involved in phenylalanine degradation in B. cepacia DBO1.
机译：在大多数细菌中，两个芳香氨基酸苯丙氨酸和酪氨酸的代谢相关。苯丙氨酸和酪氨酸分别由分支酸酯通过苯丙酮酸和对羟基苯丙酮酸合成。苯丙氨酸分解代谢的典型途径是通过苯丙氨酸初始羟基化为酪氨酸而进行的。但是，苯丙氨酸的分解代谢在某些微生物中尚不十分清楚，最近描述了一种以苯乙酸乙酸酯为中间体的苯丙氨酸降解的替代途径。洋葱伯克霍尔德菌DBO1利用单独的分解代谢途径来利用苯丙氨酸和酪氨酸。为了阐明洋葱伯劳氏菌DBO1中苯丙氨酸的降解，我们广泛使用了基于等离子的随机插入诱变来在目标分解代谢途径中生成突变体。获得了四个不能在酪氨酸上生长的突变菌株，并且得到了十个不能在苯丙氨酸上生长的菌株。有趣的是，所有酪氨酸分解代谢突变株均在苯丙氨酸上生长，表明苯丙氨酸不会通过酪氨酸降解。苯丙氨酸突变体菌株的序列分析表明，转座子插入分为三类：编码乙酸苯酯降解的操纵子，编码调节子的基因和编码丙酮酸氧化还原酶的基因。这些数据与HPLC代谢物积累研究相结合，表明苯丙氨酸的降解通过乙酸苯酯进行。通过合适的生化技术（酶法测定和代谢物鉴定）检查了预计参与分解代谢途径的蛋白质的酶功能。 RT-PCR实验用于检查洋葱伯克霍尔德菌DBO1中苯丙氨酸降解相关基因的调控。

著录项

  • 作者

    Cruz-Fisher, Maria I.;

  • 作者单位

    Rutgers The State University of New Jersey and University of Medicine and Dentistry of New Jersey.;

  • 授予单位 Rutgers The State University of New Jersey and University of Medicine and Dentistry of New Jersey.;
  • 学科 Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2006
  • 页码 94 p.
  • 总页数 94
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;
  • 关键词

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