High cost of traditional substrates and formation of by-products (such as acetone and ethanol) in acetone–butanol–ethanol (ABE) fermentation hindered the large-scale production of biobutanol. Here, we comprehensively characterized a newly isolated solventogenic and xylanolytic Clostridium species, which could produce butanol at a high ratio with elimination of ethanol and conversion of acetone to more value-added product, isopropanol. Ultimately, direct butanol production from hemicellulose was achieved with efficient expression of indigenous xylanase by the novel strain via consolidated bioprocessing. A novel wild-type Clostridium sp. strain NJP7 was isolated and characterized in this study, which was capable of fermenting monosaccharides, e.g., glucose into butanol via a fermentative acetone–isopropanol–butanol pathway. With enhancement of buffering capacity and alcohol dehydrogenase activities, butanol and isopropanol titer by Clostridium sp. strain NJP7 was improved to 12.21 and 1.92 g/L, respectively, and solvent productivity could be enhanced to 0.44 g/L/h. Furthermore, with in situ extraction with biodiesel, the amount of butanol and isopropanol was finally improved to 25.58 and 5.25 g/L in the fed-batch mode. Meanwhile, Clostridium sp. strain NJP7 shows capability of direct isopropanol–butanol production from hemicelluloses with expression of indigenous xylanase. 2.06 g/L of butanol and 0.54 g/L of isopropanol were finally achieved through the temperature-shift simultaneous saccharification and fermentation, representing the highest butanol production directly from hemicellulose. The co-production of isopropanol with butanol by the newly isolated Clostridium sp. strain NJP7 would add on the economical values for butanol fermentation. Furthermore, the high isopropanol-butanol production with in situ extraction would also greatly enhance the economic feasibility for fermentative production of butanol–isopropanol in large scale. Meanwhile, its direct production of butanol–isopropanol from polysaccharides, hemicellulose through secretion of indigenous thermostable xylanase, shows great potential using lignocellulosic wastes for biofuel production.

Galactooligosaccharides (GOS) are one of the most important functional oligosaccharide prebiotics. The surface display of enzymes was considered one of the most excellent strategies to obtain these products. However, a rough industrial environment would affect the biocatalytic process. The catalytic process could be efficiently improved using biofilm-based fermentation with high resistance and activity. Therefore, the combination of the surface display of β-galactosidase and biofilm formation in Pichia pastoris was constructed. The results showed that the catalytic conversion rate of GOS was up to 50.3% with the maximum enzyme activity of 5125 U/g by screening the anchorin, and the number of the continuous catalysis batches was up to 23 times. Thus, surface display based on biofilm-immobilized fermentation integrated catalysis and growth was a co-culture system, such that a dynamic equilibrium in the consolidated integrative process was achieved. This study provides the basis for developing biofilm-based surface display methods in P. pastoris during biochemical production processes.

The biofilm (BF) provides favorable growth conditions to cells, which has been exploited in the field of industrial biotechnology. Based on our previous research works on type I fimbriae for the biosynthesis of L-threonine (LT) in Escherichia coli, in this study, a fimA-overexpressing strain was engineered, which improved BF formation under industrial fermentation conditions. The morphological observation and characterization of BF formation were conducted to verify the function of the subunit FimA. However, it was not suitable for repeated-batch immobilized fermentation as the LT titer was not elevated significantly. The underlying molecular mechanisms of BF formation and the LT carbon flux were explored by transcriptomic analysis. The results showed that fimA regulated E. coli BF formation but affected LT carbon distribution. This study will stimulate thoughts about how the fimbriae gene regulated biofilms and amino acid excretion and will bring some consideration and provide a reference for the development of BF-based biomanufacturing processes in E. coli.

Biofilms enable bacterial cells to adhere and thrive on surfaces, with associated changes in growth and gene expression aiding their survival in challenging environments. While previous research has explored E. coli biofilm formation, there has been limited exploration of its application in industrial production. Prior studies have shown that immobilized fermentation can enhance L-threonine production. This study aims to augment biofilm formation and subsequently increase L-threonine production in E. coli by regulating the quorum sensing system, focusing on key AI-2-related genes, including luxS, lsrB, lsrK, and lsrR. In +pluxS and +plsrB strains, AI-2 levels were significantly altered, resulting in enhanced biofilm formation, increased curli expression, shorter free-cell fermentation periods, and improved production efficiency through immobilized continuous fermentation. In a single batch of free-cell fermentation with E. coli W1688, L-threonine production was 10.16 g/L. However, +pluxS and +plsrB strains achieved L-threonine yields of 15.27 g/L and 13.38 g/L, respectively, after seven fermentation batches. Additionally, the fermentation period was reduced from 36 h to 28 h and 30 h, respectively.

β-cyclodextrin (β-CD) is an important cyclic oligosaccharide, which is widely applicated in foods, environmental protection, and cosmetics, primarily prepared from enzymatic synthesis in traditional industry. However, several challenges persist, including cumbersome processes and difficulties in achieving continuous fermentation and catalysis. This research introduced a biofilm-based immobilized fermentation, integrating with enzyme catalysis system of surface display in Bacillus subtilis. The bslA gene was selected to construct the surface display system due to its ability to promote biofilm formation and serve as an anchorin. Compared to free cell catalysis, the biofilm-based immobilized catalysis expanded the temperature range to 40–70 and the pH range to 5–7.5. During the continuous catalysis process, by the 13th batch, the relative activity remained around 52%, and the conversion rate exceeded 36%, similar to the single-batch free cell catalysis. These findings provide valuable insights and effective strategies for the industrial production of β-CD and other biochemicals through continuous catalysis.