Jiajun Hu

Design and composition of synthetic fungal-bacterial microbial consortia that improve lignocellulolytic enzyme activity

Microbial interactions are important for metabolism as they can improve or reduce metabolic efficiency. To improve lignocellulolytic enzyme activity, a series of synergistic microbial consortia of increasing diversity and complexity were devised using fungal strains, including Trichoderma reesei, Penicillium decumbens, Aspergillus tubingensis, and Aspergillus niger. However, when a screened microbial community with cellulolytic capacity was added to the consortia to increase the number of strains, it engendered more microbial interactions with the above strains and universally improved the β-glucosidase activity of the consortia. Analysis of the microbial community structure revealed that the bacteria in the consortia are more important for lignocellulolytic enzyme activity than the fungi. One fungal and 16 bacterial genera in the consortia may interact with T. reesei and are potential members of a devised synergistic microbial consortium. Such devised microbial consortia may potentially be applied to effectively and economically degrade lignocellulose.

Influence of rice straw polyphenols on cellulase production by Trichoderma reesei

In this study, we found that during cellulase production by Trichoderma reesei large amounts of polyphenols were released from rice straw when the latter was used as the carbon source. We identified and quantified the phenolic compounds in rice straw and investigated the effects of the phenolic compounds on cellulase production by T.xa0reesei. The phenolic compounds of rice straw mainly consisted of phenolic acids and tannins. Coumaric acid (CA) and ferulic acid (FA) were the predominant phenolic acids, which inhibited cellulase production by T.xa0reesei. When the concentrations of CA and FA in the broth increased to 0.06xa0g/L, cellulase activity decreased by 23% compared with that in the control culture. Even though the rice straw had a lower tannin than phenolic acid content, the tannins had a greater inhibitory effect than the phenolic acids on cellulase production by T.xa0reesei. Tannin concentrations greater than 0.3xa0g/L completely inhibited cellulase production. Thus, phenolic compounds, especially tannins are the major inhibitors of cellulase production by T.xa0reesei. Therefore, we studied the effects of pretreatments on the release of phenolic compounds. Ball milling played an important role in the release of FA and CA, and hot water extraction was highly efficient in removing tannins. By combining ball milling with extraction by water, the 2-fold higher cellulase activity than in the control culture was obtained.

Effects of different cellulases on the release of phenolic acids from rice straw during saccharification

Effects of different cellulases on the release of phenolic acids from rice straw during saccharification were investigated in this study. All cellulases tested increased the contents of phenolic acids during saccharification. However, few free phenolic acids were detected, as they were present in conjugated form after saccharification when the cellulases from Trichoderma reesei, Trichoderma viride and Aspergillus niger were used. On the other hand, phenolic acids were present in free form when the Acremonium cellulolyticus cellulase was used. Assays of enzyme activity showed that, besides high cellulase activity, the A. cellulolyticus cellulase exhibited high feruloyl esterase (FAE) activity. A synergistic interaction between FAE and cellulase led to the increase in free phenolic acids, and thus an increase in antioxidative and antiradical activities of the phenolic acids. Moreover, a cost estimation demonstrated the feasibility of phenolic acids as value-added products to reduce the total production cost of ethanol.

Response of cbb gene transcription levels of four typical sulfur-oxidizing bacteria to the CO2 concentration and its effect on their carbon fixation efficiency during sulfur oxidation

The variability in carbon fixation capability of four sulfur-oxidizing bacteria (Thiobacillus thioparus DSM 505, Halothiobacillus neapolitanus DSM 15147, Starkeya novella DSM 506, and Thiomonas intermedia DSM 18155) during sulfur oxidation was studied at low and high concentrations of CO2. The mechanism underlying the variability in carbon fixation was clarified by analyzing the transcription of the cbb gene, which encodes the key enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase. DSM 15147 and DSM 505 fixed carbon more efficiently during sulfur oxidation than DSM 506 and DSM 18155 at 0.5% and 10% CO2, which was mainly because their cbb gene transcription levels were much higher than those of DSM 506 and DSM 18155. A high CO2 concentration significantly stimulated the carbon fixation efficiency of DSM 505 by greatly increasing the cbb gene transcription efficiency. Moreover, the influence of the CO2 concentration on the carbon fixation efficiency of the four strains differed greatly during sulfur oxidation.

Release of Polyphenols Is the Major Factor Influencing the Bioconversion of Rice Straw to Lactic Acid

In this study, we found that p-coumaric acid (p-CA), ferulic acid (FA), and condensed tannins were released from rice straw during saccharification. The presence of polyphenols prolonged the lag phase and lowered the productivity of lactic acid. p-CA was identified as a key inhibitor. Tannins had a lower inhibitory effect than p-CA; FA had little inhibitory effect. Acid, alkaline, and ball milling pretreatments elicited different levels of polyphenol release from rice straw. Due to the different levels of polyphenol release in the pretreatment step, the enzymatic hydrolysates contained different concentrations of polyphenols. Compared with fermentation with a synthetic medium, fermentation with the hydrolysates of ball-milled rice straw provided much lower productivity and yield of lactic acid due to the presence of polyphenols. Removal of these compounds played an important role in lactic acid fermentation. When rice straw was alkaline pretreated, the hydrolysates contained few phenolic compounds, resulting in high productivity and yield of lactic acid (1.8xa0g/L/h and 26.7xa0g/100xa0g straw), which were comparable to those in a synthetic medium. This indicates that there is a correlation between removal of phenolic compounds and efficiency in lactic acid fermentation.

Characterization of a designed synthetic autotrophic–heterotrophic consortia for fixing CO2 without light

Microbial interactions are important for metabolism, and they improve metabolic substrate types and metabolic efficiency. To discover microbial combinations with high CO2 fixation efficiencies, a series of synergistic microbial consortia of increasing diversity and complexity were devised using chemoautotrophic strains, including Ochrobactrum, Stenotrophomonas, Castellaniella, and Sinomicrobium strains, which were isolated from a non-photosynthetic microbial community (NPMC) with CO2 fixation capacity. Addition of a small inocula of NPMC universally improved the CO2 fixation efficiencies of the consortia by up to 10-fold, while the CO2 fixation efficiencies of most multimember consortia were similar to those of single strains. An analysis of the microbial community structure revealed that both autotrophic–autotrophic microbial interactions and autotrophic–heterotrophic microbial interactions occurred in the synthetic microbial consortia. Ochrobactrum and Castellaniella strains were crucial for autotrophic metabolism, while Lysinibacillus and Pseudomonas strains were crucial for heterotrophic metabolism. These devised microbial consortia have potential applications in addressing environmental issues.

Influence of rice straw-derived dissolved organic matter on lactic acid fermentation by Rhizopus oryzae

Rice straw can be used as carbon sources for lactic acid fermentation. However, only a small amount of lactic acid is produced even though Rhizopus oryzae can consume glucose in rice straw-derived hydrolysates. This study correlated the inhibitory effect of rice straw with rice straw-derived dissolved organic matter (DOM). Lactic acid fermentations with and without DOM were conducted to investigate the effect of DOM on lactic acid fermentation by R.xa0oryzae. Fermentation using control medium with DOM showed a similar trend to fermentation with rice straw-derived hydrolysates, showing that DOM contained the major inhibitor of rice straw. DOM assay indicated that it mainly consisted of polyphenols and polysaccharides. The addition of polyphenols and polysaccharides derived from rice straw confirmed that lactic acid fermentation was promoted by polysaccharides and significantly inhibited by polyphenols. The removal of polyphenols also improved lactic acid production. However, the loss of polysaccharides during the removal of polyphenols resulted in low glucose consumption. This study is the first to investigate the effects of rice straw-derived DOM on lactic acid fermentation by R.xa0oryzae. The results may provide a theoretical basis for identifying inhibitors and promoters associated with lactic acid fermentation and for establishing suitable pretreatment methods.

Use of magnetic powder to effectively improve the performance of sequencing batch reactors (SBRs) in municipal wastewater treatment

This study aims to investigate the effect of adding magnetic powder in the sequencing batch reactor (SBR) on the reactor performance and microbial community. Results indicated that, the magnetic activated sludge sequencing batch reactor (MAS-SBR) had 7.76% and 4.76% higher ammonia nitrogen (NH4+-N) and chemical oxygen demand (COD) removal efficiencies than that of the conventional SBR (C-SBR). The MAS-SBR also achieved 6.86% sludge reduction compared with the C-SBR. High-throughput sequencing demonstrated that the dominant phyla of both SBRs (present as ≥1% of the sequence reads) were Protebacteria, Bacteroidetes, Chloroflexi, Saccharibacteria, Chlorobi, Firmicutes, Actinobactoria, Acidobacteria, Planctomycetes and unclassified_Bacteria. The relative abundance of Protebacteria and Bacteroidetes simultaneously declined whereas the other 8 phyla increased following the addition of magnetic powder. Adding magnetic powder in the SBR significantly affected the microbial diversity and richness of activated sludge, consequently affecting the reactor performance.

Inhibitory effects of phenolic compounds of rice straw formed by saccharification during ethanol fermentation by Pichia stipitis

In this study, it was found that the type of phenolic acids derived from rice straw was the major factor affecting ethanol fermentation by Pichia stipitis. The aim of this study was to investigate the inhibitory effect of phenolic acids on ethanol fermentation with rice straw. Different cellulases produced different ratios of free phenolic acids to soluble conjugated phenolic acids, resulting in different fermentation efficiencies. Free phenolic acids exhibited much higher inhibitory effect than conjugated phenolic acids. The flow cytometry results indicated that the damage to cell membranes was the primary mechanism of inhibition of ethanol fermentation by phenolic acids. The removal of free phenolic acids from the hydrolysates increased ethanol productivity by 2.0-fold, indicating that the free phenolic acids would be the major inhibitors formed during saccharification. The integrated process for ethanol and phenolic acids may constitute a new strategy for the production of low-cost ethanol.

Interactions Between Autotrophic and Heterotrophic Strains Improve CO2 Fixing Efficiency of Non-photosynthetic Microbial Communities

Five autotrophic strains isolated from non-photosynthetic microbial communities (NPMCs), which were screened from oceans with high CO2 fixing capability, were identified as Ochrobactrum sp. WH-2, Stenotrophomonas sp. WH-11, Ochrobactrum sp. WH-13, Castellaniella sp. WH-14, and Sinomicrobium oceani WH-15. The CO2 fixation pathways of all these strains were Calvin-Benson-Bassham pathway. These strains could metabolize multifarious organic compounds, which allowed switching them to autotrophic culture after enrichment in heterotrophic culture. The central composite response surface method indicated that these strains possessed many interactive effects, which increased the CO2 fixing efficiency of a combined community composed of these strains by 56xa0%, when compared with that of the single strain. Furthermore, another combined community composed of these autotrophic strains and NPMC had richer interactive relationships, with CO2 fixing efficiency being 894xa0% higher than that of the single strain and 148xa0% higher than the theoretical sum of the CO2 fixing efficiency of each of its microbial components. The interaction between strictly heterotrophic bacteria in NPMC and isolated autotrophic strains played a crucial role in improving the CO2 fixing efficiency, which not only eliminated self-restraint of organic compounds generated during the growth of autotrophic bacteria but also promoted its autotrophic pathway.