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首页> 外文期刊>Applied Microbiology >Microbial Gutta-Percha Degradation Shares Common Steps with Rubber Degradation by Nocardia nova SH22a
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Microbial Gutta-Percha Degradation Shares Common Steps with Rubber Degradation by Nocardia nova SH22a

机译：诺卡氏菌新星SH22a降解橡胶的胶质与橡胶降解有共同的步骤

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Nocardia nova SH22a, a bacterium capable of degrading gutta-percha (GP) and natural rubber (NR), was used to investigate the GP degradation mechanism and the relations between the GP and NR degradation pathways. For this strain, a protocol of electroporation was systematically optimized, and an efficiency of up to 4.3 × 10~(7) CFU per μg of plasmid DNA was achieved. By applying this optimized protocol to N. nova SH22a, a Tn 5096 -based transposon mutagenesis library of this bacterium was constructed. Among about 12,000 apramycin-resistant transformants, we identified 76 stable mutants defective in GP or NR utilization. Whereas 10 mutants were specifically defective in GP utilization, the growth of the other 66 mutants was affected on both GP and NR. This indicated that the two degradation pathways are quite similar and share many common steps. The larger number of GP-degrading defective mutants could be explained in one of two ways: either (i) the GP pathway is more complex and harbors more specific steps or (ii) the steps for both pathways are almost identical, but in the case of GP degradation there are fewer enzymes involved in each step. The analysis of transposition loci and genetic studies on interesting genes confirmed the crucial role of an α-methylacyl-coenzyme A racemase in the degradation of both GP and NR. We also demonstrated the probable involvement of enzymes participating in oxidoreduction reactions, β-oxidation, and the synthesis of complex cell envelope lipids in the degradation of GP.
机译：能够降解牙胶（GP）和天然橡胶（NR）的诺卡氏菌诺卡氏菌诺氏菌SH22a被用于研究GP的降解机理以及GP和NR降解途径之间的关系。对于该菌株，系统地优化了电穿孔方案，每μg质粒DNA的效率高达4.3×10〜（7）CFU。通过将该优化方案应用于新星猪SH22a，构建了该细菌的基于Tn 5096的转座子诱变文库。在大约12,000个抗阿霉素霉素的转化子中，我们鉴定出76个稳定的GP或NR利用缺陷突变体。尽管有10个突变体在GP利用方面存在特定缺陷，但其他66个突变体的生长同时受到GP和NR的影响。这表明这两个降解途径非常相似，并且共有许多共同步骤。大量GP降解缺陷突变体可以通过以下两种方式之一进行解释：（i）GP途径更复杂且包含更具体的步骤，或者（ii）两种途径的步骤几乎相同，但在这种情况下由于GP降解，每个步骤涉及的酶较少。对转座基因座的分析和对有趣基因的遗传研究证实了α-甲基酰基辅酶A消旋酶在GP和NR降解中的关键作用。我们还证明了GP降解中酶可能参与氧化还原反应，β-氧化和复杂细胞膜脂质的合成。

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