Liu Yan, Ding Xiu-Min, Xue Zheng-Lian, Hu Liu-Xiu, Zhang Ning-Juan, Wang Zhou, Yang Jian-Wei, Cheng Qian, Chen Ming-Hong, Zhang Zhuang-Zhuang, Zheng Zhi-Ming
College of Biochemical Engineering, Anhui Polytechnic University, Wuhu, 241000, China.
Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Science, Hefei, 230031, China.
World J Microbiol Biotechnol. 2017 Mar;33(3):52. doi: 10.1007/s11274-017-2222-9. Epub 2017 Feb 14.
Menaquinone (MK) was an attractive membrane-bound intracellular chemical. To enhance its production, we tried to find the relationship between its synthesis and the state of cell membrane in producing strain. Due to non-ionic surfactant-polyoxyethylene oleyl ether (POE) and plant oil-cedar wood oil (CWO) can typically increase extracellular secretion and intracellular synthesis of MK respectively, the effect of these two substances on cell morphology, physical properties of cell membrane was investigated. Finally, two engineering strains were constructed to verify whether the state of cell membrane can enhance MK synthesis. The result showed that the edge of cells was broken when POE added in the medium. Other physical properties such as total fatty acid content decreased by 40.7% and the ratio of saturated fatty acids to unsaturated fatty acids decreased from 1.58 ± 0.05 to 1.31 ± 0.04. Meanwhile, cell membrane leakage was enhanced from 7.14 to 64.31%. Different from POE group, cell membrane was intact in CWO group. Moreover, the ratio of saturated fatty acids to unsaturated fatty acids increased from 1.58 ± 0.05 to 1.78 ± 0.04 and the average lipid length decreased from 16.05 ± 0.08 to 15.99 ± 0.10. Two constructed strains, especially Escherichia coli DH5α FatB, exhibited strong MK secretion ability and the extracellular MK reached 10.71 ± 0.19 mg/L. An understanding of these functionary mechanisms could not only provide a new idea for the synthesis of MK, but also provide a reference to increase the yield of intracellular membrane-bound metabolites.
甲萘醌(MK)是一种具有吸引力的膜结合细胞内化学物质。为提高其产量,我们试图找出其合成与生产菌株细胞膜状态之间的关系。由于非离子表面活性剂聚氧乙烯油醚(POE)和植物油雪松油(CWO)通常分别能增加MK的细胞外分泌和细胞内合成,因此研究了这两种物质对细胞形态、细胞膜物理性质的影响。最后,构建了两株工程菌株以验证细胞膜状态是否能增强MK的合成。结果表明,在培养基中添加POE时,细胞边缘破裂。其他物理性质如总脂肪酸含量下降了40.7%,饱和脂肪酸与不饱和脂肪酸的比例从1.58±0.05降至1.31±0.04。同时,细胞膜泄漏率从7.14%提高到64.31%。与POE组不同,CWO组细胞膜完整。此外,饱和脂肪酸与不饱和脂肪酸的比例从1.58±0.05增加到1.78±0.04,平均脂质长度从16.05±0.08降至15.99±0.10。构建的两株菌株,尤其是大肠杆菌DH5α FatB,表现出较强的MK分泌能力,细胞外MK达到10.71±0.19 mg/L。对这些作用机制的了解不仅可为MK的合成提供新思路,也可为提高细胞内膜结合代谢产物的产量提供参考。