Background: The co-fermentation of syngas (mainly CO, H 2 and CO 2) and different concentrations of carbohydrate/ protein synthetic wastewater to produce volatile fatty acids (VFAs) was conducted in the present study. Results: It was found that co-fermentation of syngas with carbohydrate-rich synthetic wastewater could enhance the conversion efficiency of syngas and the most efficient conversion of syngas was obtained by co-fermentation of syngas with 5 g/L glucose, which resulted in 25% and 43% increased conversion efficiencies of CO and H 2 , compared to syngas alone. The protein-rich synthetic wastewater as co-substrate, however, had inhibition on syngas conversion due to the presence of high concentration of NH 4 +-N (> 900 mg/L) produced from protein degradation. qPCR analysis found higher concentration of acetogens, which could use CO and H 2 , was present in syngas and glucose co-fermentation system, compared to glucose solo-fermentation or syngas solo-fermentation. In addition, the known acetogen Clostridium formicoaceticum, which could utilize both carbohydrate and CO/H 2 was enriched in syngas solofermentation and syngas with glucose co-fermentation. In addition, butyrate was detected in syngas and glucose co-fermentation system, compared to glucose solo-fermentation. The detected n-butyrate could be converted from acetate and lactate/ethanol which produced from glucose in syngas and glucose co-fermentation system supported by label-free quantitative proteomic analysis. Conclusions: These results demonstrated that the co-fermentation with syngas and carbohydrate-rich wastewater could be a promising technology to increase the conversion of syngas to VFAs. In addition, the syngas and glucose co-fermentation system could change the degradation pathway of glucose in co-fermentation and produce fatty acids with longer carbon chain supported by microbial community and label-free quantitative proteomic analysis. The above results are innovative and lead to achieve effective conversion of syngas into VFAs/longer chain fatty acids, which would for sure have a great interest for the scientific and engineering community. Furthermore, the present study also used the combination of high-throughput sequencing of 16S rRNA genes, qPCR analysis and label-free