Construction of stable microbial consortia for effective biochemical synthesis.

Microbial consortia can complete otherwise arduous tasks through the cooperation of multiple microbial species. This concept has been applied to produce commodity chemicals, natural products, and biofuels. However, metabolite incompatibility and growth competition can make the microbial composition unstable, and fluctuating microbial populations reduce the efficiency of chemical production. Thus, controlling the populations and regulating the complex interactions between different strains are challenges in constructing stable microbial consortia. This Review discusses advances in synthetic biology and metabolic engineering to control social interactions within microbial cocultures, including substrate separation, byproduct elimination, crossfeeding, and quorum-sensing circuit design. Additionally, this Review addresses interdisciplinary strategies to improve the stability of microbial consortia and provides design principles for microbial consortia to enhance chemical production.

Zinc glycerolate (Glyzinc): A novel foliar fertilizer for zinc biofortification and cadmium reduction in wheat (Triticum aestivum L.).

Sustainable strategies are essential for zinc (Zn) biofortification and cadmium (Cd) reduction in staple food crops. Herein, we evaluated the phytotoxicity of Glyzinc under foliar and root application (FA&RA) in a lab-scale experiment, and then investigated its Zn efficiency and Cd reduction through foliar application on wheat (Triticum aestivum L.) under field conditions. Compared to RA, FA of Glyzinc exhibited no adverse effect on wheat growth and oxidative stresses at all doses. In field conditions, FA of Glyzinc remarkably increased Zn (28.7 %), S (10.4 %), Cu (17.3 %) and crude protein (9.1 %) content in wheat grain at 100 mg/L without damaging wheat yield. Furthermore, FA of Glyzinc significantly reduced the grain phytic acid (PA) (23.7 %) and Cd level (19.5 %), as well as PA to Zn molar ratio (32.3 %). Overall, our results indicate that Glyzinc has great potential as a high-efficiency foliar fertilizer for Zn biofortification and safe crop production in nano-enabled agriculture.

Molecular composition of soil organic matter (SOM) regulate qualities of tobacco leaves.

Soil organic matter (SOM) is of vital importance to soil health, and also plays a crucial role in the quality of the crops such as tobacco. However, the link between tobacco quality and SOM chemical compositions is still not well understood. To fill the information gap, we analyzed the quality of tobacco leaves and the corresponding SOM molecular compositions by electrospray ionization (ESI) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), that were collected from six different sites in Bijie, Guizhou Province, China. The tobacco quality variedin six sites based on their chemical compositions. SOM compounds had a remarked impact on the quality of tobacco leaves and a distinct difference in SOM composition between low-quality and high-quality tobacco leaves was observed as well. Specifically, 105 common molecular formulas were detected in three SOM compounds of high-quality tobacco, which were more than those in low-quality samples. Although amino sugar, proteins, lipids, tannins, and carbohydrates had a collective influence on the chemical composition of tobacco leaves, the effect contributed by amino sugar and tannins was more prominent. In summary, fully understanding the association between tobacco chemical composition and SOM compounds can provide new insight into the regulation of tobacco quality and the sustainable development of agriculture.

Synthesis of acidified magnetic sludge-biochar and its role in ammonium nitrogen removal: Perception on effect and mechanism.

An acidified magnetic sludge-biochar (MSB) was prepared to enhance ammonium nitrogen (AN) removal efficiency in eutrophic water, and MSB was obtained by secondary pyrolysis of sludge biochar powder. A series of MSB were prepared under 300, 400, 500, 600 °C and different valence states of iron ions by impregnation pyrolysis, which is based on the deposition of unstable iron minerals on biochar matrix. Physicochemical properties of pristine biochar and MSB were revealed through characterization analysis, suggesting that MSB prepared by ferric chloride at 400 °C presented the largest adsorption capacity, and the acid-modification enhanced the ammonium adsorption capacity by 10.7%. Electrostatic attraction and ion-exchange processes were identified as the main adsorption mechanisms of MSB on AN. As the most dominant mechanism, ion exchange of AN with functional groups containing -OH and CO on the surface of MSB resulted in the relative content of -OH (61.3%) and CO (11.5%) bonds reduced to 34.2% and 7.0% respectively. The novel magnetic sludge-biochar with acid-modification possessed enhanced electron transfer capacity, revealing a removal pathway of ammonium by nitrification. The findings above demonstrated that MSB is a promising adsorbent for ammonium removal and can be applied to the natural nitrogen-rich water regulation.

Quantitative proteomic analysis to reveal expression differences for butanol production from glycerol and glucose by Clostridium sp. strain CT7.

Clostridium sp. strain CT7 is a new emerging microbial cell factory with high butanol production ratio owing to its non-traditional butanol fermentation mode with uncoupled acetone and 1,3-propanediol formation. Significant changes of metabolic products profile were shown in glycerol- and glucose-fed strain CT7, especially higher butanol and lower volatile fatty acids (VFAs) production occurred from glycerol-fed one. However, the mechanism of this interesting phenomenon was still unclear. To better elaborate the bacterial response towards glycerol and glucose, the quantitative proteomic analysis through iTRAQ strategy was performed to reveal the regulated proteomic expression levels under different substrates. Proteomics data showed that proteomic expression levels related with carbon metabolism and solvent generation under glycerol media were highly increased. In addition, the up-regulation of hydrogenases, ferredoxins and electron-transferring proteins may attribute to the internal redox balance, while the earlier triggered sporulation response in glycerol-fed media may be associated with the higher butanol production. This study will pave the way for metabolic engineering of other industrial microorganisms to obtain efficient butanol production from glycerol.

Spatio-temporally expressed sorbitol transporters cooperatively regulate sorbitol accumulation in pear fruit.

Sorbitol is the primary substrate translocated from source to sink in pear species. Among the many sorbitol transporters (SOTs), some are known to be involved in sorbitol accumulation in fruit; however, their particular roles are unclear. In this study, we examined the transcriptome and metabolome of a variety of pear samples from six time points to identify those SOTs. Similar to previous studies, sorbitol and sucrose differed significantly between the leaf and fruit, and sorbitol was consistently observed at higher concentrations at all time points. Interestingly, we found that sorbitol accumulation in pear fruit was cooperatively mediated by SOT3, SOT6/20, SOT19/21, and SOT22. In particular, the up-regulated SOT6/20 and SOT19/21 in fruit under 1 mg L-1 abscisic acid and 10 mg L-1 indole acetic acid treatments, respectively, resulted in an increased sorbitol concentration. In addition, sorbitol concentration showed positive correlations to fructose and glucose concentrations, indicating a role for sorbitol in the determination of fruit sweetness. Together with the deduced process of sugar biosynthesis, transport, conversion, and accumulation in pear, our study provides a foundation for further research into sugar accumulation processes in pear fruit, contributing to the improvement of fruit quality.

Consolidated bioprocessing performance of a two-species microbial consortium for butanol production from lignocellulosic biomass.

Consolidated bioprocessing (CBP) by using microbial consortium was considered as a promising approach to achieve direct biofuel production from lignocellulose. In this study, the interaction mechanism of microbial consortium consisting of Thermoanaerobacterium thermosaccharolyticum M5 and Clostridium acetobutylicum NJ4 was analyzed, which could achieve efficient butanol production from xylan through CBP. Strain M5 possesses efficient xylan degradation capability, as 19.73 g/L of xylose was accumulated within 50 hr. The efficient xylose utilization capability of partner strain NJ4 could relieve the substrate inhibition to hydrolytic enzymes of xylanase and xylosidase secreted by strain M5. In addition, the earlier solventogenesis of strain NJ4 was observed due to the existence of butyrate generated by strain M5. The mutual interaction of these two strains finally gave 13.28 g/L of butanol from 70 g/L of xylan after process optimization, representing a relatively high butanol production from hemicellulose. Moreover, 7.61 g/L of butanol was generated from untreated corncob via CBP. This successfully constructed microbial consortium exhibits efficient cooperation performance on butanol production from lignocellulose, which could provide a platform for the emerging butanol production from lignocellulose.

Recent advances of biofuels and biochemicals production from sustainable resources using co-cultivation systems.

Microbial communities are ubiquitous in nature and exhibit several attractive features, such as sophisticated metabolic capabilities and strong environment robustness. Inspired by the advantages of natural microbial consortia, diverse artificial co-cultivation systems have been metabolically constructed for biofuels, chemicals and natural products production. In these co-cultivation systems, especially genetic engineering ones can reduce the metabolic burden caused by the complex of metabolic pathway through labor division, and improve the target product production significantly. This review summarized the most up-to-dated co-cultivation systems used for biofuels, chemicals and nature products production. In addition, major challenges associated with co-cultivation systems are also presented and discussed for meeting further industrial demands.

Automatic Cell Segmentation by Adaptive Thresholding (ACSAT) for Large-Scale Calcium Imaging Datasets.

Advances in calcium imaging have made it possible to record from an increasingly larger number of neurons simultaneously. Neuroscientists can now routinely image hundreds to thousands of individual neurons. An emerging technical challenge that parallels the advancement in imaging a large number of individual neurons is the processing of correspondingly large datasets. One important step is the identification of individual neurons. Traditional methods rely mainly on manual or semimanual inspection, which cannot be scaled for processing large datasets. To address this challenge, we focused on developing an automated segmentation method, which we refer to as automated cell segmentation by adaptive thresholding (ACSAT). ACSAT works with a time-collapsed image and includes an iterative procedure that automatically calculates global and local threshold values during successive iterations based on the distribution of image pixel intensities. Thus, the algorithm is capable of handling variations in morphological details and in fluorescence intensities in different calcium imaging datasets. In this paper, we demonstrate the utility of ACSAT by testing it on 500 simulated datasets, two wide-field hippocampus datasets, a wide-field striatum dataset, a wide-field cell culture dataset, and a two-photon hippocampus dataset. For the simulated datasets with truth, ACSAT achieved >80% recall and precision when the signal-to-noise ratio was no less than ∼24 dB.

Heterologous expression of cyclodextrin glycosyltransferase from Paenibacillus macerans in Escherichia coli and its application in 2-O-α-D-glucopyranosyl-L-ascorbic acid production.

BACKGROUND: Cyclodextrin glucanotransferase (CGTase) can transform L-ascorbic acid (L-AA, vitamin C) to 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G), which shows diverse applications in food, cosmetic and pharmaceutical industries. RESULTS: In this study, the cgt gene encoding α-CGTase from Paenibacillus macerans was codon-optimized (opt-cgt) and cloned into pET-28a (+) for intracellular expression in E. coli BL21 (DE3). The Opt-CGT was purified by Ni2+-NTA resin with a 55% recovery, and specific activity was increased significantly from 1.17 to 190.75 U·mg- 1. In addition, the enzyme was adopted to transform L-AA into 9.1 g/L of AA-2G. Finally, more economic substrates, including ß-cyclodextrin, soluble starch, corn starch and cassava starch could also be used as glycosyl donors, and 4.9, 3.5, 1.3 and 1.5 g/L of AA-2G were obtained, respectively. CONCLUSIONS: N-terminal amino acid is critical to the activity of CGTase suggested by its truncation study. Furthermore, when the Opt-CGT was flanked by His6-tags on the C- and N-terminal, the recovery of purification by Ni2+-NTA resin is appreciably enhanced. α-cyclodextrin was the ideal glycosyl donor for AA-2G production. In addition, the selection of low cost glycosyl donors would make the process of AA-2G production more economically competitive.