Xiaoyu Zhu

The synthesis of n -caproate from lactate: a new efficient process for medium-chain carboxylates production

A unique microbiome that metabolizes lactate rather than ethanol for n-caproate production was obtained from a fermentation pit used for the production of Chinese strong-flavour liquor (CSFL). The microbiome was able to produce n-caproate at concentrations as high as 23.41u2009g/L at a maximum rate of 2.97u2009g/L/d in batch trials without in-line extraction. Compared with previous work using ethanol as the electron donor, the n-caproate concentration increased by 82.89%. High-throughput sequencing analysis showed that the microbiome was dominated by a Clostridium cluster IV, which accounted for 79.07% of total reads. A new process for n-caproate production was proposed, lactate oxidation coupled to chain elongation, which revealed new insight into the well-studied lactate conversion and carbon chain elongation. In addition, these findings indicated a new synthesis mechanism of n-caproate in CSFL. We believe that this efficient process will provide a promising opportunity for the innovation of waste recovery as well as for n-caproate biosynthesis.

Enhanced methane production in an anaerobic digestion and microbial electrolysis cell coupled system with co-cultivation of Geobacter and Methanosarcina

The anaerobic digestion (AD) and microbial electrolysis cell (MEC) coupled system has been proved to be a promising process for biomethane production. In this paper, it was found that by co-cultivating Geobacter with Methanosarcina in an AD-MEC coupled system, methane yield was further increased by 24.1%, achieving to 360.2 mL/g-COD, which was comparable to the theoretical methane yield of an anaerobic digester. With the presence of Geobacter, the maximum chemical oxygen demand (COD) removal rate (216.8 mg COD/(L·hr)) and current density (304.3A/m(3)) were both increased by 1.3 and 1.8 fold compared to the previous study without Geobacter, resulting in overall energy efficiency reaching up to 74.6%. Community analysis demonstrated that Geobacter and Methanosarcina could coexist together in the biofilm, and the electrochemical activities of both were confirmed by cyclic voltammetry. Our study observed that the carbon dioxide content in total gas generated from the AD reactor with Geobacter was only half of that generated from the same reactor without Geobacter, suggesting that Methanosarcina may obtain the electron transferred from Geobacter for the reduction of carbon dioxide to methane. Taken together, Geobacter not only can improve the performance of the MEC system, but also can enhance methane production.

Production of high-concentration n-caproic acid from lactate through fermentation using a newly isolated Ruminococcaceae bacterium CPB6

Backgroundn-Caproic acid (CA), as a medium-chain carboxylic acid, is a valuable chemical feedstock for various industrial applications. The fermentative production of CA from renewable carbon sources has attracted a lot of attentions. Lactate is a significant intermediate waste in the anaerobic breakdown of carbohydrates that comprise 18–70% of the chemical oxygen demand (COD) in municipal and some industrial wastewaters. Recently, researchers (including our own group) reported the CA production using lactate as electron donor with newly identified microbiome systems. However, within such processes, it was hard to determine whether the CA production was completed by a single strain or by the co-metabolism of different microorganisms.ResultsHere, we report the CA production using lactate as electron donor using the strain CPB6, which we isolated from a microbiome for CA production as described previously. Strain CPB6 is affiliated with Clostridium cluster IV of the family of Ruminococcaceae based on 16S rRNA gene sequence analysis. The strain prefers acidic initial pH condition (pH 5.0–6.5), and the temperature ranging from 30 to 40xa0°C for CA production. In a fed-batch fermentation with non-sterilized lactate-containing organic wastewater as feedstock, strain CPB6 produced 16.6xa0g/L CA (from 45.1xa0g/L lactate) with a maximum productivity of 5.29xa0g/L/day. Enzyme assays with crude cell extract showed that CPB6 can metabolize acetate and butyryl-CoA to produce n-butyric acid, and acetate/n-butyrate and caproyl-CoA to produce CA, respectively.ConclusionThis study demonstrated that high concentration of CA production can be obtained by a newly isolated pure culture CPB6. This strain can be employed as a powerful workhorse for high-efficient CA recovery from lactate-containing waste streams. Our preliminary investigation suggested that the CA production from lactate in strain CPB6 might be via the chain elongation pathway of the reverse β-oxidation; the detailed mechanism, however, warrants further investigation using various molecular microbiology techniques.

Production of Butyrate from Lactate by a Newly Isolated Clostridium sp. BPY5

Lactate-utilizing bacteria play important roles in the production of Chinese strong-flavored liquor (CSFL). However, the identity of these bacteria and their lactate-utilizing properties are largely unknown. Here, a lactate-utilizing, butyrate-producing bacterium BPY5 was isolated from an old fermentation pit for CSFL production. The isolate represented a novel species belonging to Clostridium cluster XIVa of family Lachnospiraceae based on phylogenetic analysis using 16S rRNA gene sequences. Strain BPY5 could ferment lactate into butyrate as the major metabolic product. Butyrate was significantly formed at initial lactate concentration from 66 to 104xa0mM, but substantially declined when initial lactate exceeded 133xa0mM. At initial lactate concentration of 66xa0mM, lactate conversion was independent on initial pH from 5.5 to 7.0, but the conversion was completely inhibited when pH dropped below 4.8. Nevertheless, the inhibition on lactate conversion was largely relieved by the addition of acetate, suggesting that exogenous acetate could enhance lactate conversion at low pH condition. Additionally, lactate in CSFL-brewing wastewater was dramatically removed when inoculated with strain BPY5. These results implicate that the isolate may be applied for the industrial production of butyrate or the recovery of butyrate from lactate-containing wastewater.

The functional potential and active populations of the pit mud microbiome for the production of Chinese strong-flavour liquor

The popular distilled Chinese strong‐flavour liquor (CSFL) is produced by solid fermentation in the ground pit. Microbes inhabiting in the pit mud (PM) on the walls of the fermentation pit are responsible for the production of caproic acid (CA) that determines the quality of CSFL to a large degree. However, little is known about the active microbial populations and metabolic potential of the PM microbiome. Here, we investigated the overall metabolic features of the PM microbiome and its active microbial components by combining metagenomics and MiSeq‐sequencing analyses of the 16S rRNA genes from DNA and RNA (cDNA). Results showed that prokaryotes were predominant populations in the PM microbiome, accounting for 95.3% of total metagenomic reads, while eukaryotic abundance was only 1.8%. The dominant prokaryotic phyla were Firmicutes, Euryarchaeota, Bacteroidetes, Actinobacteria and Proteobacteria, accounting for 48.0%, 19.0%, 13.5%, 2.5% and 2.1% of total metagenomic reads respectively. Most genes encoding putative metabolic pathways responsible for the putative CA production via chain elongation pathway were detected. This indicated that the PM microbiome owned functional potential for synthesizing CA from ethanol or lactate. Some key genes encoding enzymes involved in hydrogenotrophic and acetoclastic methanogenesis pathways were detected in the PM metagenome, suggesting the possible occurrence of interspecies hydrogen transfer between CA‐producing bacteria and methanogens. The 16S rDNA and 16S rRNA profiles showed that the Clostridial cluster IV, Lactobacillus, Caloramator, Clostridium, Sedimentibacter, Bacteroides and Porphyromonas were active populations in situ, in which Clostridial cluster IV and Clostridium were likely involved in the CA production. This study improved our understandings on the active populations and metabolic pathways of the PM microbiome involved in the CA synthesis in the CSFL fermentation.

Complete genome sequence of Ruminococcaceae bacterium CPB6: A newly isolated culture for efficient n-caproic acid production from lactate

n-caproic acid (CA) is a valuable chemical feedstock for various industrial applications. Biological production of CA from renewable carbon sources has attracted a lot of attentions recently. We lately reported the new culture Ruminococcaceae bacterium CPB6, which was isolated from a microbiome for efficient CA production from lactate. To further elucidate its metabolism, we sequenced the whole genome of the strain. The size of the complete genome is 2,069,994bp with 50.58% GC content; no plasmid was identified. Sets of genes involved in the fatty acid biosynthesis via acyl carrier protein (ACP) and coenzyme A (CoA) as well as lactate oxidation/reduction pathways were identified in the genome. These genes were inferred to be correlated with the CA production. The complete genome sequence provides essential information for the elucidation of the metabolism for CA production from lactate, and further improvement of the strain through genetic engineering for enhanced CA production and other biotechnological purposes.

The process-related dynamics of microbial community during a simulated fermentation of Chinese strong-flavored liquor

BackgroundFamous Chinese strong-flavored liquor (CSFL) is brewed by microbial consortia in a special fermentation pit (FT). However, the fermentation process was not fully understood owing to the complicate community structure and metabolism. In this study, the process-related dynamics of microbial communities and main flavor compounds during the 70-day fermentation process were investigated in a simulated fermentation system.ResultsA three-phase model was proposed to characterize the process of the CSFL fermentation. (i) In the early fermentation period (1–23xa0days), glucose was produced from macromolecular carbohydrates (e.g., starch). The prokaryotic diversity decreased significantly. The Lactobacillaceae gradually predominated in the prokaryotic community. In contrast, the eukaryotic diversity rose remarkably in this stage. Thermoascus, Aspergillus, Rhizopus and unidentified Saccharomycetales were dominant eukaryotic members. (ii) In the middle fermentation period (23–48xa0days), glucose concentration decreased while lactate acid and ethanol increased significantly. Prokaryotic community was almost dominated by the Lactobacillus, while eukaryotic community was mainly comprised of Thermoascus, Emericella and Aspergillus. (iii) In the later fermentation period (48–70xa0days), the concentrations of ethyl esters, especially ethyl caproate, increased remarkably.ConclusionsThe CSFL fermentation could undergo three stages: saccharification, glycolysis and esterification. Saccharomycetales, Monascus, and Rhizopus were positively correlated to glucose concentration (Pxa0<xa00.05), highlighting their important roles in the starch saccharification. The Lactobacillaceae, Bacilli, Botryotinia, Aspergillus, unidentified Pleosporales and Capnodiales contributed to the glycolysis and esterification, because they were positively correlated to most organic acids and ethyl esters (Pxa0<xa00.05). Additionally, four genera, including Emericella, Suillus, Mortierella and Botryotinia, that likely played key roles in fermentation, were observed firstly. This study observed comprehensive dynamics of microbial communities during the CSFL fermentation, and it further revealed the correlations between some crucial microorganisms and flavoring chemicals (FCs). The results from this study help to design effective strategies to manipulate microbial consortia for fermentation process optimization in the CSFL brew practice.

Multiferroic properties of Bi0.5K0.5TiO3–BiFe1−xCoxO3 (0 ≤ x ≤ 0.2) solid solution

The fabrication of BiFeO3 and Bi0.5K0.5TiO3 typically encounters problems with densification and phase purity, requiring conditions of high pressure for the synthesis of bulk phases. In this letter, we have successfully prepared a binary lead-free solid-solution of Bi0.5K0.5TiO3–BiFe1−xCoxO3 using a modified Pechini method and investigated the magnetic and ferroelectric properties of Bi0.5K0.5TiO3–BiFe1−xCoxO3 (0 ≤ x ≤ 0.2). The coexistence of room-temperature ferromagnetism and ferroelectricity is observed in Bi0.5K0.5TiO3–BiFe1−xCoxO3 (0.5 ≤ x ≤ 0.2). The x = 0.2 sample exhibits ferromagnetic behavior with a Curie temperature TC of 661 K, compared with the paramagnetic like behavior exhibited in Bi0.5K0.5TiO3–BiFeO3 at room temperature. The ferromagnetism can be ascribed to the suppression of a spiral spin structure with the canting of the anti-ferromagnetically ordered spins caused by structural distortion due to the substitution of Co ions. The x = 0.2 sample also exhibits a well-defined ferroelectric hysteresis loop with a rather large remnant polarization (Pr = 32.72 μC cm−2), which is superior to that of other lead-free ferroelectric compounds. The improved ferroelectric properties can be attributed to the increased grain size and lattice distortion caused by Co doping. The observation of room temperature ferromagnetism and ferroelectricity for Bi0.5K0.5TiO3–BiFe1−xCoxO3 suggests that these materials are potential candidates for multiferroic materials and ferroelectric memory devices.

Lead ions removal from aqueous solution in a novel bioelectrochemical system with a stainless steel cathode

Heavy metal pollution, especially lead pollution in water, has been a growing concern due to the toxicity of lead to human and other beings. According to previous reports, bioelectrochemical systems (BESs) showed significant advantages in heavy metal ions removal, but have not been considered for Pb2+ removal. In this study, a novel BES with stainless steel cathode distinguished with traditional BESs was employed with mixed culture as biocatalyst for removing Pb2+ from solution. The results indicated Pb2+ could be effectively removed and hydrocerussite as the final product confirmed by X-ray diffraction was deposited on the stainless steel cathode. Furthermore, the principle of Pb2+ removal was deduced based on the experiment of the reduction of ferricyanide in the stainless steel tube-type BES. In brief, we suggested a novel low-cost approach to remove and recover Pb2+ from Pb2+-containing wastewater.

Promotion of Ni2+ Removal by Masking Toxicity to Sulfate-Reducing Bacteria: Addition of Citrate

The sulfate-reducing bioprocess is a promising technology for the treatment of heavy metal-containing wastewater. This work was conducted to investigate the possibility of promoting heavy metal removal by the addition of citrate to mask Ni2+ toxicity to sulfate-reducing bacteria (SRB) in batch reactors. SRB growth was completely inhibited in Ni2+-containing medium (1 mM) when lactate served as the sole carbon resource, leading to no sulfate reduction and Ni2+ removal. However, after the addition of citrate, SRB grew well, and sulfate was quickly reduced to sulfide. Simultaneously, the Ni-citrate complex was biodegraded to Ni2+ and acetate. The NiS precipitate was then formed, and Ni2+ was completely removed from the solution. It was suggested that the addition of citrate greatly alleviates Ni2+ toxicity to SRB and improves the removal of Ni2+, which was confirmed by quantitative real-time PCR targeting dissimilatory sulfite reductase (dsrAB) genes. Analysis of the carbon metabolism indicated that lactate instead of acetate served as the electron donor for sulfate reduction. This study offers a potential approach to increase the removal of heavy metals from wastewater in the single stage SRB-based bioprocess.