Wang Peng, Chen Hao, Wei Xin, Liu Guang-Lei, Chi Zhe, Jiang Bo, Chi Zhen-Ming
College of Marine Life Sciences, Ocean University of China, Yushan Road, No. 5, Qingdao, China.
Laboratory for Marine Biology and Biotechnology, Qingdao Marine Science and Technology Center, Qingdao, China.
Microb Cell Fact. 2024 Dec 4;23(1):327. doi: 10.1186/s12934-024-02608-6.
Xylose from lignocellulose is one of the most abundant and important renewable and green raw materials. It is very important how to efficiently transform xylose into useful bioproducts such as fumaric acid and so on.
In this study, it was found that the GC1 strain (∆gox, in which the GOX gene encoding glucose oxidase which could transform glucose into gluconic acid was removed) of A. pullulans var. aubasidani DH177 had the high ability to utilize xylose and corncob-derived xylose with CO fixation derived from CaCO to produce calcium fumarate. Overexpression of the XI gene encoding xylose isomerase, the XK gene encoding xylose kinase and the TKL gene coding for transketolase made the strain TKL-4 produce 73.1 g/L of calcium fumarate from xylose. At the same time, the transcriptional levels of the key ASS gene coding for argininosuccinate synthase and the ASL gene coding for argininosuccinate lyase in the ornithine-urea cycle (OUC) were also obviously enhanced. The results also demonstrated that the TKL-4 strain could produce more calcium fumarate from xylose and corncob-derived xylose than from glucose. During 10-liter fermentation, the TKL-4 strain could produce 88.5 g/L of calcium fumarate from xylose, the productivity was 0.52 g/h/L. Meanwhile, it could yield 85.6 g/L of calcium fumarate from corncob-derived xylose and the productivity was 0.51 g/h/L. During the same fermentation, the TKL-4 strain could transform the mixture containing 75.0 g/L glucose and 45.0 g/L xylose to produce 78.7 ± 1.1 g/L calcium fumarate.
This indicated that the TKL-4 strain constructed in this study indeed could actively transform xylose and corncob-derived xylose into calcium fumarate through the green ways.
木质纤维素中的木糖是最丰富且重要的可再生绿色原料之一。如何将木糖高效转化为富马酸等有用的生物产品至关重要。
本研究发现,普鲁兰短梗霉DH177的GC1菌株(∆gox,其中编码可将葡萄糖转化为葡萄糖酸的葡萄糖氧化酶的GOX基因被去除)具有利用木糖和玉米芯衍生木糖以及利用碳酸钙固定的CO来生产富马酸钙的高能力。编码木糖异构酶的XI基因、编码木糖激酶的XK基因和编码转酮醇酶的TKL基因的过表达使TKL - 4菌株从木糖中生产出73.1 g/L的富马酸钙。同时,鸟氨酸 - 尿素循环(OUC)中编码精氨琥珀酸合酶的关键ASS基因和编码精氨琥珀酸裂解酶的ASL基因的转录水平也明显提高。结果还表明,TKL - 4菌株从木糖和玉米芯衍生木糖中生产的富马酸钙比从葡萄糖中生产的更多。在10升发酵过程中,TKL - 4菌株从木糖中可生产88.5 g/L的富马酸钙,生产率为0.52 g/h/L。同时,它从玉米芯衍生木糖中可生产85.6 g/L的富马酸钙,生产率为0.51 g/h/L。在相同发酵过程中,TKL - 4菌株可将含有75.0 g/L葡萄糖和45.0 g/L木糖的混合物转化为生产78.7 ± 1.1 g/L的富马酸钙。
这表明本研究构建的TKL - 4菌株确实能够通过绿色途径将木糖和玉米芯衍生木糖积极转化为富马酸钙。