Key Laboratory of Molecular Microbiology Technology, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, China.
Tianjin Engineering Technology Center of Green Manufacturing Biobased Materials, Tianjin, China.
Microbiologyopen. 2019 Feb;8(2):e00631. doi: 10.1002/mbo3.631. Epub 2018 Apr 15.
In the industrial production of xanthan gum using Xanthomonas campestris CGMCC15155, large amounts of ethanol are required to extract xanthan gum from the fermentation broth and remove xanthomonadin impurities. To reduce the amount of ethanol and the overall production cost of xanthan gum, a xanthomonadin-deficient strain of CGMCC15155 was constructed by inserting the Vitreoscilla globin (vgb) gene, under the control of the LacZ promoter, into the region of the pigA gene, which is involved in xanthomonadin synthesis. The insertion of vgb inactivated pigA, resulting in the production of white xanthan gum. The lack of xanthomonadins resulted in a decreased yield of xanthan gum. However, the expression product of vgb gene, VHb, could increase the metabolism of X. campestris, which allowed the production of xanthan gum to reach wild-type levels in the engineered strain. The yield, molecular weight, and rheological properties of the xanthan gum synthesized by the engineered and wild-type bacteria were essentially the same. When the same volume of ethanol was used, the whiteness values of the xanthan gum extracted from engineered and wild-type bacteria were 65.20 and 38.17, respectively. To extract xanthan gum with the same whiteness, three and seven times the fermentation volume of ethanol was required for the engineered and wild-type strains, respectively. Thus, the engineered train reduced the requirement for ethanol in xanthan gum production by 133.3%. The results demonstrated that the engineered bacteria used less ethanol, thus reducing the downstream processing cost in xanthan gum production.
在利用野油菜黄单胞菌 CGMCC15155 进行黄原胶的工业生产中,需要大量的乙醇从发酵液中提取黄原胶并去除黄原胶毒素杂质。为了减少乙醇的用量和黄原胶的整体生产成本,通过将编码玻璃珠蛋白(Vitreoscilla globin,VHb)的 vgb 基因插入到参与黄原胶毒素合成的 pigA 基因区域,构建了一株黄原胶毒素缺陷型 CGMCC15155 突变株。vgb 的插入失活了 pigA,导致产生了白色黄原胶。黄原胶毒素的缺失导致黄原胶产量降低。然而,vgb 基因的表达产物 VHb 可以增加野油菜黄单胞菌的代谢,使得工程菌中黄原胶的产量达到野生型水平。工程菌和野生型细菌合成的黄原胶的产量、分子量和流变性能基本相同。当使用相同体积的乙醇时,从工程菌和野生型细菌中提取的黄原胶的白度值分别为 65.20 和 38.17。为了提取具有相同白度的黄原胶,工程菌和野生型菌株分别需要发酵液体积的 3 倍和 7 倍的乙醇。因此,工程菌将黄原胶生产中对乙醇的需求减少了 133.3%。结果表明,该工程菌减少了乙醇的使用量,从而降低了黄原胶生产的下游加工成本。