Improving stress tolerance and cell integrity of Rhodococcus ruber by overexpressing small-shock-protein Hsp16 of Rhodococcus
Improving stress tolerance and cell integrity of Rhodococcus ruber by overexpressing small-shock-protein Hsp16 of Rhodococcus
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DOI：
10.1007/s10295-018-2066-9
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发表时间：
2018-07
影响因子：
3.4
通讯作者：
Miaomiao Wang;Jie Chen;Huimin Yu;Zhongyao Shen
中科院分区：
文献类型：
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作者：
Miaomiao Wang;Jie Chen;Huimin Yu;Zhongyao Shen
红球菌属的物种已被成功地用作有价值的化学品生产的细胞催化剂，由于其良好的表征有害因素的抵抗力。了解它们如何应对压力是非常有意义的，这将有助于确定进一步提高其抵抗力和保持细胞完整性和活力的工程策略。在这里，我们评估了R. ruberTH 3热休克。大约有376个基因在热休克TH 3中上调。在所有上调的功能基因中，确定了转录增强最大（463倍）的小热休克蛋白（Hsp 16），并对其功能进行了研究。结果表明，过量表达Hsp 16对胞内酶的抗逆性没有明显的促进作用。有趣的是，与对照TH 3相比，AM处理的TH 3（Hsp 16）细胞表面上的孔和褶皱略少，而TH 3（Hsp-GFP）细胞表面上的完整性更高。存活试验表明，50%AM处理后，TH 3（Hsp 16）菌落数增加（约501-700），而只有1-100个，高温培养实验也发现了这一趋势。这些结果表明，Hsp 16在防止细胞渗漏、维持细胞完整性和细胞活力方面具有重要作用。橡胶在压力条件下。
Rhodococcusspecies have been successfully used as cell catalysts for valuable chemicals production due to their well-characterized resistance to harmful factors. An understanding of how they respond to stress is of great interest, which will enable the identification of engineering strategies for further improving their resistance and maintaining cell integrity and viability. Here, we assessed the transcriptome response ofR. ruberTH3 to heat shock. Approximately, 376 genes were up-regulated in heat-shocked TH3. Among all the up-regulated functional genes, the small heat-shock-protein (Hsp16) with maximal enhanced transcript (463-fold) was identified, and its function was investigated. Results showed that overexpressed Hsp16 has no significant promotive effect on stress tolerance of in-cell enzyme. Interestingly, compared to the control TH3, a little fewer pores and folds on the surface of TH3(Hsp16) and more intact TH3(Hsp-GFP) cells under AM treatment were observed by SEM and LCSM, respectively. Moreover, survival test showed that more (about 501–700) TH3(Hsp16) colonies were observed while only 1–100 TH3 colonies after 50% AM treatment, and this trend is also found in high-temperature cultivation experiments. These results indicate that Hsp16 does great contributions to preventing cell leakage, maintaining cell integrity and viability ofR. ruberunder stress conditions.