Vongkampang Thitiwut, Sreenivas Krishnan, Engvall Jonathan, Grey Carl, van Niel Ed W J
Division of Applied Microbiology, Lund University, P.O. Box 124, 221 00, Lund, Sweden.
Division of Biotechnology, Lund University, P.O. Box 124, 221 00, Lund, Sweden.
Biotechnol Biofuels. 2021 Apr 8;14(1):91. doi: 10.1186/s13068-021-01938-6.
Caldicellulosiruptor kronotskyensis has gained interest for its ability to grow on various lignocellulosic biomass. The aim of this study was to investigate the growth profiles of C. kronotskyensis in the presence of mixtures of glucose-xylose. Recently, we characterized a diauxic-like pattern for C. saccharolyticus on lignocellulosic sugar mixtures. In this study, we aimed to investigate further whether C. kronotskyensis has adapted to uptake glucose in the disaccharide form (cellobiose) rather than the monosaccharide (glucose).
Interestingly, growth of C. kronotskyensis on glucose and xylose mixtures did not display diauxic-like growth patterns. Closer investigation revealed that, in contrast to C. saccharolyticus, C. kronotskyensis does not possess a second uptake system for glucose. Both C. saccharolyticus and C. kronotskyensis share the characteristics of preferring xylose over glucose. Growth on xylose was twice as fast (μ = 0.57 h) as on glucose (μ = 0.28 h). A study of the sugar uptake was made with different glucose-xylose ratios to find a kinetic relationship between the two sugars for transport into the cell. High concentrations of glucose inhibited xylose uptake and vice versa. The inhibition constants were estimated to be K = 0.01 cmol L and K = 0.001 cmol L, hence glucose uptake was more severely inhibited by xylose uptake. Bioinformatics analysis could not exclude that C. kronotskyensis possesses more than one transporter for glucose. As a next step it was investigated whether glucose uptake by C. kronotskyensis improved in the form of cellobiose. Indeed, cellobiose is taken up faster than glucose; nevertheless, the growth rate on each sugar remained similar.
C. kronotskyensis possesses a xylose transporter that might take up glucose at an inferior rate even in the absence of xylose. Alternatively, glucose can be taken up in the form of cellobiose, but growth performance is still inferior to growth on xylose. Therefore, we propose that the catabolism of C. kronotskyensis has adapted more strongly to pentose rather than hexose, thereby having obtained a specific survival edge in thermophilic lignocellulosic degradation communities.
克氏热纤梭菌因其能够在各种木质纤维素生物质上生长而受到关注。本研究的目的是调查克氏热纤梭菌在葡萄糖 - 木糖混合物存在下的生长曲线。最近,我们表征了嗜糖热纤梭菌在木质纤维素糖混合物上的类似二次生长模式。在本研究中,我们旨在进一步研究克氏热纤梭菌是否已适应摄取二糖形式(纤维二糖)而非单糖(葡萄糖)的葡萄糖。
有趣的是,克氏热纤梭菌在葡萄糖和木糖混合物上的生长未显示出类似二次生长的模式。进一步研究发现,与嗜糖热纤梭菌不同,克氏热纤梭菌不具有第二个葡萄糖摄取系统。嗜糖热纤梭菌和克氏热纤梭菌都具有优先利用木糖而非葡萄糖的特征。在木糖上的生长速度(μ = 0.57 h⁻¹)是在葡萄糖上(μ = 0.28 h⁻¹)的两倍。研究了不同葡萄糖 - 木糖比例下的糖摄取情况,以找到两种糖转运进入细胞的动力学关系。高浓度的葡萄糖抑制木糖摄取,反之亦然。抑制常数估计为K = 0.01 cmol L⁻¹和K = 0.001 cmol L⁻¹,因此木糖摄取对葡萄糖摄取的抑制作用更强。生物信息学分析不能排除克氏热纤梭菌拥有不止一种葡萄糖转运蛋白的可能性。接下来研究了克氏热纤梭菌以纤维二糖形式摄取葡萄糖的情况是否有所改善。事实上,纤维二糖的摄取速度比葡萄糖快;然而,每种糖上的生长速率仍然相似。
克氏热纤梭菌拥有一种木糖转运蛋白,即使在没有木糖的情况下,它摄取葡萄糖的速率可能也较低。或者,葡萄糖可以以纤维二糖的形式被摄取,但生长性能仍低于在木糖上的生长。因此,我们提出克氏热纤梭菌的分解代谢对戊糖的适应性比对己糖更强,从而在嗜热木质纤维素降解群落中获得了特定的生存优势。