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通过敲除与嗜盐古菌胞外多糖生物合成相关的基因来提高聚羟基烷酸酯的产量。

Improving polyhydroxyalkanoate production by knocking out the genes involved in exopolysaccharide biosynthesis in Haloferax mediterranei.

机构信息

State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Number 1 Beichen West Road, Chaoyang District, Beijing, 100101, China.

出版信息

Appl Microbiol Biotechnol. 2013 Apr;97(7):3027-36. doi: 10.1007/s00253-012-4415-3. Epub 2012 Sep 27.

Abstract

Haloferax mediterranei is capable of producing large amounts of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) from many kinds of carbon sources, with exopolysaccharide (EPS) as a by-product. In this study, we identified a gene cluster involved in EPS biosynthesis in H. mediterranei. Knocking out the genes in this cluster encoding the putative UDP-N-acetylglucosamine 6-dehydrogenase (HFX_2145), glycosyltransferases (HFX_2146 and HFX_2147) and polysaccharide transporter (HFX_2148) eliminated EPS synthesis. The deficiency in EPS biosynthesis in the mutant strain remarkably decreased the viscosity of culture broth, and hence increased the dissolved oxygen content and decreased the foaming propensity. Compared with the wild-type (WT) strain, the PHBV production of the EPS-mutant strain was significantly enhanced (approximately 20%), whereas the cell growth rate remained similar under the same culture conditions. These results indicated that the carbon sources used for synthesizing EPS were shifted to PHBV production. Thus, a novel engineered H. mediterranei strain was developed, which would be favorable for future industrial production of PHBV.

摘要

地中海盐杆菌能够利用多种碳源大量生产聚(3-羟基丁酸酯-共-3-羟基戊酸酯)(PHBV),同时副产物是胞外多糖(EPS)。在本研究中,我们鉴定了地中海盐杆菌中参与 EPS 生物合成的基因簇。敲除该簇中编码假定的 UDP-N-乙酰葡萄糖胺 6-脱氢酶(HFX_2145)、糖基转移酶(HFX_2146 和 HFX_2147)和多糖转运蛋白(HFX_2148)的基因,会消除 EPS 的合成。突变株中 EPS 生物合成的缺陷显著降低了发酵液的粘度,从而增加了溶解氧含量并降低了起泡倾向。与野生型(WT)菌株相比,EPS 突变株的 PHBV 产量显著提高(约 20%),而在相同的培养条件下,细胞生长速率保持相似。这些结果表明,用于合成 EPS 的碳源被转移到 PHBV 的生产上。因此,开发了一种新型工程化的地中海盐杆菌菌株,这将有利于未来 PHBV 的工业生产。

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