Prabhu Ashish A, Ledesma-Amaro Rodrigo, Lin Carol Sze Ki, Coulon Frederic, Thakur Vijay Kumar, Kumar Vinod
School of Water, Energy and Environment, Cranfield University, Cranfield, MK43 0AL UK.
Department of Bioengineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, SW7 2AZ UK.
Biotechnol Biofuels. 2020 Jun 27;13:113. doi: 10.1186/s13068-020-01747-3. eCollection 2020.
Xylose is the most prevalent sugar available in hemicellulose fraction of lignocellulosic biomass (LCB) and of great interest for the green economy. Unfortunately, most of the cell factories cannot inherently metabolize xylose as sole carbon source. is a non-conventional yeast that produces industrially important metabolites. The yeast is able to metabolize a large variety of substrates including both hydrophilic and hydrophobic carbon sources. However, lacks effective metabolic pathway for xylose uptake and only scarce information is available on utilization of xylose. For the economica feasibility of LCB-based biorefineries, effective utilization of both pentose and hexose sugars is obligatory.
In the present study, succinic acid (SA) production from xylose by was examined. To this end, PSA02004 strain was engineered by overexpressing pentose pathway cassette comprising xylose reductase (), xylitol dehydrogenase () and xylulose kinase () gene. The recombinant strain exhibited a robust growth on xylose as sole carbon source and produced substantial amount of SA. The inhibition of cell growth and SA formation was observed above 60 g/L xylose concentration. The batch cultivation of the recombinant strain in a bioreactor resulted in a maximum biomass concentration of 7.3 g/L and SA titer of 11.2 g/L with the yield of 0.19 g/g. Similar results in terms of cell growth and SA production were obtained with xylose-rich hydrolysate derived from sugarcane bagasse. The fed-batch fermentation yielded biomass concentration of 11.8 g/L (OD: 56.1) and SA titer of 22.3 g/L with a gradual decrease in pH below 4.0. Acetic acid was obtained as a main by-product in all the fermentations.
The recombinant strain displayed potential for bioconversion of xylose to SA. Further, this study provided a new insight on conversion of lignocellulosic biomass into value-added products. To the best of our knowledge, this is the first study on SA production by using xylose as a sole carbon source.
木糖是木质纤维素生物质(LCB)半纤维素部分中最普遍存在的糖类,对绿色经济具有重要意义。不幸的是,大多数细胞工厂无法天然地将木糖作为唯一碳源进行代谢。[酵母名称]是一种能产生具有工业重要性代谢产物的非常规酵母。该酵母能够代谢多种底物,包括亲水性和疏水性碳源。然而,[酵母名称]缺乏有效的木糖摄取代谢途径,关于木糖利用的信息也很稀少。对于基于LCB的生物精炼厂的经济可行性而言,有效利用戊糖和己糖都是必不可少的。
在本研究中,检测了[酵母名称]利用木糖生产琥珀酸(SA)的情况。为此,通过过表达包含木糖还原酶([酶名称1])、木糖醇脱氢酶([酶名称2])和木酮糖激酶([酶名称3])基因的戊糖途径盒对PSA02004菌株进行了工程改造。重组菌株在以木糖为唯一碳源的培养基上表现出强劲的生长,并产生了大量的SA。当木糖浓度高于60 g/L时,观察到细胞生长和SA形成受到抑制。在生物反应器中对重组菌株进行分批培养,得到最大生物量浓度为7.3 g/L,SA产量为11.2 g/L,产率为0.19 g/g。使用源自甘蔗渣的富含木糖的水解产物,在细胞生长和SA生产方面也得到了类似的结果。补料分批发酵得到生物量浓度为11.8 g/L(OD:56.1),SA产量为22.3 g/L,pH值逐渐降至4.0以下。在所有发酵过程中,乙酸是主要副产物。
重组菌株显示出将木糖生物转化为SA的潜力。此外,本研究为将木质纤维素生物质转化为高附加值产品提供了新的见解。据我们所知,这是首次关于[酵母名称]利用木糖作为唯一碳源生产SA的研究。