 Hi, this is He Jinzong from the Department of Civil and Environmental Engineering at the National University of Singapore. The research interest of my lab is to explore the anaerobic microbial world to make discoveries that advance scientific understanding of mechanisms and mechanisms responsible for bio-transformation and fermentation of environmental pollutants and biomass residues. While my research topic focuses on converting biomass to biochemicals to meet the ever-increasing demands for cleaner biochemicals and biofuels, over the past several years, my research group has been working on discovering new strengths that hold promise in generating several biochemical and biofuel related products from different carbon sources such as food waste, starch, and ligno-synallocic biomass. However, utilization of ligno-synallocic biomass being composed of both 5 and 6 carbon sugars is challenging, considering the fact that subgenic bacteria in general prefer to utilize 6-carbon sugar only. The major breakthrough in this regard was obtained in my lab with the discovery of a strain-built stream which can simultaneously utilize both 5 and 6-carbon sugars. Strain-built stream is particularly interesting since it produces a similar amount of butanol from both glucose and zealots. Particularly for zealots, it produces a significantly larger amount of riboflavin, another commercially important product. To conduct in-depth understanding about the proteomics of stream BOH3, we collaborated with Dr. Lee Chin-Sung and his team as a protein at Proteomics Center at the National University of Singapore. The subsequent experimental design along with some major findings, which is a subject of our upcoming paper in biotechnology and bioengineering, would now be discussed by Dr. Basu. Hello, my name is Dr. Basu and I am going to talk about our upcoming paper as told by Dr. Herb. As already discussed, apart from its capability to utilize both 5 and 6 carbon sugars, the most important aspect about our strain-BH3 was its capability to produce riboflavin at higher titers, whilst maintaining similar butanol eats and as zealots fit conditions. Availability of a commercially important entity like riboflavin as a by-product is particularly interesting from bioprocessed economics point of view, which urged us to pursue an in-depth understanding of the biology of the bacteria. An nitrate-based proteomics investigation was therefore performed with this bacteria at different stages of fermentation. Samples were taken at different time points from the fermentation model, late ascetic phase and mid-solventic phase. In this study, an influx of high-tractate was used to label the bacteria proteins as shown in this figure, such that different high-tract ratios could eat valuable information like the expression ratio of proteins at different stages of fermentation, whilst using gluobos and zealots as the carbon source, as well as reveal substrate-specific proteomic dynamics. Through our academic experiments, we were able to quantify expression ratios of approximately 20% of the total proteome of the bacterial cell as shown in Figure 4. The quantified proteins were found to be well-distributed within all classes of the COC classification, indicating the possibility of capturing detailed metabolic changes within the bacteria through our experimental results. However, since our focus in this study was to understand the missionaries involved in phytozaptic anti-cochlorine production, we focused on certain classes of proteins for the sake of simplicity. A closer study of the COC proteins related to energy metabolism indicated the optimization of several plug-up proteins within the valence-fed pH-3 cells. Alongside, we also observed some proteins represented as the top cluster C3 in this figure, which were down-regulated in the zealots-fed crocultures due to solvent proteomic phase. Interestingly, we found that this cluster consisted of proteins belonging to the electron transport chain of the bacteria, which is further supported by the flow-side electric data of the samples, where we could observe a significant bacterial population to be metabolically inactive due to the loss of the electron transport chain function. With regard to the riboflavin production, we also found all proteins related to riboflavin production partly to be upregulated within the zealots-fed cultures as shown in Figure 6. Interestingly, riboflavin production between the cells decreased with increase in aerosine concentration within the culture media, indicating that riboflavin produced by the bacteria is actually involved in iron uptake. Coming back to proteins involved in zealots utilization, we found two distinct operands to be upregulated within the zealots-fed cultures, namely CAC-1341-249 and CAC-2610-213. The CAC-1341-249 operand seems to be particularly important since it contains a zealots-fed quota. This operand, however, remains under the negative control of the CCPA protein, represented as CAC-3037 over here, which is a known catapulted repressor protein producing pluripotent functions within the cellular metabolism. Contrary to the other studies related to clostridium fermentation, this protein did not show any signs of regulation between the glucose and zealots-fed cultures, thereby indicating that it remains rigidly suppressed under zealots-fed conditions. Another important observation for the case of zealots-fed cultures is the upregulation of several iron-containing proteins related to redox stress, particularly in the zealots-fed phase. Simultaneously, we also observed the regulation of several correlation-related genes within the zealots-fed bacteria at its zealots-fed phase. Our observations thus indicate that strain-BH3 is highly capable of uplaking zealots and regulating its metabolic machineries when exposed to the 5-carbon sugar owing to the abdomen-fixed repression of the inhibitory CCPA protein. Such a metabolic setup helps to efficiently utilize zealots on the one hand. On the other hand, it leads to increased iron demands leading to higher lipoclubric production under various restricted conditions. Such metabolic consequences can ultimately result in increased cellular-oxidated stress, which can be detrimental particularly during the salventilating phase when the bacteria are already being exposed to gradually increasing cell stress. Hence, we propose that future metabolic engineering studies of cost-reduced strains should focus on methods to manage the stresses within the target of carousels so as to improve internal production on the one hand and the workflow in the other. For further details regarding the study, we can refer to our paper in Biotechnology and Bioengineering. Hope you enjoyed reading it. Happy reading!