| 1株转座子插入突变菌株TB34的筛选及产氢分析 |
| 摘要点击 1519 全文点击 1153 投稿时间：2011-09-29 修订日期：2011-11-10 |
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| 中文关键词 转座子 Tn7 成团泛菌 碳源 产氢量 |
| 英文关键词 transposon Tn7 Pantoea agglomerans BH18 carbon source hydrogen production |
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| 中文摘要 |
| 以分离自红树林污泥的厌氧发酵产氢细菌Pantoea agglomerans BH18为出发菌株,利用转座子Tn7构建突变体文库.通过卡那霉素抗性筛选与PCR扩增,鉴定转座子插入突变菌株.通过初筛和复筛,获得1株突变菌TB34,其产氢量较野生菌株明显提高.在初始pH为7.0和葡萄糖浓度10 g·L-1的海水培养条件下,产氢量(H2/葡萄糖)为(2.04±0.04)mol·mol-1,相比野生菌株产氢量提高43%.经过5次连续传代培养,突变菌株TB34表现出稳定的产氢特性.测定突变菌株TB34在不同碳源培养条件下的产氢量.结果表明,突变菌株TB34和野生菌株BH18都能利用蔗糖、葡萄糖和果糖发酵产氢.与野生菌株BH18不同,突变菌株TB34在以木糖为底物培养条件下仍能够发酵产氢,产氢量(H2/木糖)为(1.34±0.09)mol·mol-1,扩大了底物利用范围. |
| 英文摘要 |
| To increase the hydrogen-producing capacity of Pantoea agglomerans BH18, isolated from mangrove sludge, we constructed a stable transposon mutagenesis library of this strain. A Tn7-based transposon was randomly inserted into the genomic DNA. Mutants were screened by kanamycin resistance and identified by amplification of the inserted transposon sequences. A mutant strain TB34 was isolated, whose hydrogen production capacity was significantly improved compared to the wild type strain. In seawater-containing medium supplemented with 10 g·L-1 glucose and had an initial pH of 7.0, the hydrogen yield (H2/glucose) of the mutant strain was (2.04±0.04) mol·mol-1, which was 43% higher than that of the wild type strain. The mutant TB34 showed steady hydrogen production capacity for five consecutive passages. Different carbon sources were tested in the hydrogen production by the mutant TB34 and the results showed that both the mutant strain TB34 and the wild type strain BH18 were able to produce hydrogen on sucrose, glucose and fructose. However, different from the wild type strain, the mutant strain TB34 was also able to produce hydrogen using xylose as substrate, with a hydrogen yield (H2/xylose) of (1.34±0.09) mol·mol-1, indicating a broader substrate spectrum in the mutant. |
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