The rapid industrial and economic development runs on fossil fuel and other energy sources. Limited oil reserves, environmental issues, and high transportation costs lead towards carbon unbiased renewable and sustainable fuel. Compared to other carbon-based fuels, biodiesel is attracted worldwide as a biofuel for the reduction of global dependence on fossil fuels and the greenhouse effect. During biodiesel production, approximately 10% of glycerol is formed in the transesterification process in a biodiesel plant. The ditching of crude glycerol is important as it contains salt, free fatty acids, and methanol that cause contamination of soil and creates environmental challenges for researchers. However, the excessive cost of crude glycerol refining and market capacity encourage the biodiesel industries for developing a new idea for utilising and produced extra sources of income and treat biodiesel waste. This review focuses on the significance of crude glycerol in the value-added utilisation and conversion to bioethanol by a fermentation process and describes the opportunities of glycerol in various applications.

/pdf/glycerol-waste-to-value-added-products-and-its-potential-cp7t51me8o.pdf

Glycerol waste to value added products and its potential applications

Reprogramming complex cellular metabolism requires simultaneous regulation of multigene expression. Ex-situ cloning-based methods are commonly used, but the target gene number and combinatorial library size are severely limited by cloning and transformation efficiencies. In-situ methods such as multiplex automated genome engineering (MAGE) depends on high-efficiency transformation and incorporation of heterologous DNA donors, which are limited to few microorganisms. Here, we describe a Base Editor-Targeted and Template-free Expression Regulation (BETTER) method for simultaneously diversifying multigene expression. BETTER repurposes CRISPR-guided base editors and in-situ generates large numbers of genetic combinations of diverse ribosome binding sites, 5’ untranslated regions, or promoters, without library construction, transformation, and incorporation of DNA donors. We apply BETTER to simultaneously regulate expression of up to ten genes in industrial and model microorganisms Corynebacterium glutamicum and Bacillus subtilis. Variants with improved xylose catabolism, glycerol catabolism, or lycopene biosynthesis are respectively obtained. This technology will be useful for large-scale fine-tuning of multigene expression in both genetically tractable and intractable microorganisms. To obtain optimal yield and productivity in bioproduction, expression of pathway genes must be appropriately coordinated. Here, the authors report repurposing of base editors for simultaneous regulation of multiple gene expression and demonstrate its application in industrially important and model microorganisms.

/pdf/in-situ-generation-of-large-numbers-of-genetic-combinations-3wk7ettdbh.pdf

In-situ generation of large numbers of genetic combinations for metabolic reprogramming via CRISPR-guided base editing

The continuous temperature rise has raised global concerns about CO2 emissions. As the country with the largest CO2 emissions, China is facing the challenge of achieving large CO2 emission reductions (or even net-zero CO2 emissions) in a short period. With the strong support and encouragement of the Chinese government, technological breakthroughs and practical applications of carbon capture, utilization, and storage (CCUS) are being aggressively pursued, and some outstanding accomplishments have been realized. Based on the numerous information from a wide variety of sources including publications and news reports only available in Chinese, this paper highlights the latest CCUS progress in China after 2019 by providing an overview of known technologies and typical projects, aiming to provide theoretical and practical guidance for achieving net-zero CO2 emissions in the future.

/pdf/a-review-of-recent-progress-of-carbon-capture-utilization-3cl8ptka.pdf

A Review of Recent Progress of Carbon Capture, Utilization, and Storage (CCUS) in China

Engineering of Corynebacterium glutamicum for high-level γ-aminobutyric acid production from glycerol by dynamic metabolic control.

Highly efficient production of L-homoserine in Escherichia coli by engineering a redox balance route.

Construction of a carbon-conserving pathway for glycolate production by synergetic utilization of acetate and glucose in Escherichia coli

Glucose and xylose are two major components of lignocellulose. Simultaneous consumption of glucose and xylose is critical for engineered microorganisms to produce fuels and chemicals from lignocellulosic biomass. Although many production limitations have been resolved, glucose-induced inhibition of xylose transport remains a challenge. In this study, a cell growth-based screening strategy was designed to identify xylose transporters uninhibited by glucose. The glucose pathway was genetically blocked in Escherichia coli so that glucose functions only as an inhibitor and cells need xylose as the carbon source for survival. Through adaptive evolution, omics analysis and reverse metabolic engineering, a new phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS) galactitol transporter (GalABC, encoded by EcolC_1640, EcolC_1641, and EcolC_1642 genes) that is not inhibited by glucose was identified. Inactivation of adenylate cyclase led to increased expression of the EcolC_1642 gene, and a point mutation in gene EcolC_1642 (N13S) further enhanced xylose transport. During the second round of gene mining, AraE and a new ABC transporter (AraFGH) of xylose were identified. A point mutation in the transcription regulator araC (L156I) caused increased expression of araE and araFGH genes without arabinose induction, and a point mutation in araE (D223Y) further enhanced xylose transport. These newly identified xylose transporters can support the simultaneous consumption of glucose and xylose and have potential use in producing chemicals from lignocellulose. 

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/mlf2.12021

New xylose transporters support the simultaneous consumption of glucose and xylose in
 Escherichia coli

Construction of Escherichia coli cell factories

The invention discloses recombinant bacteria for producing succinic acid with glycerin being a substrate and a construction method and application thereof. In order to reduce impurities and increase the yield of succinic acid, an II-type glycerin metabolism system is introduced into Escherichia coli, molecular modification is conducted on the recombinant bacteria Suc-T11, an electronic transfer chain working under an anaerobic condition is constructed, the supply capacity of energy is improved, and then the yield of succinic acid is increased. Experiments verify that in batched fermentation, the yield of succinic acid in the recombinant bacteria YY-GS004 reaches 482.5 mM and is 3.82 times that of a parent, and the conversion rate reaches 0.92 mol succinic acid/mol glycerin, reaches 92% ofa theoretical value and approximates to a theoretical conversion rate; in supplementing fermentation adopting an aerobiotic-anaerobic two-step method, the yield of succinic acid reaches 565.3 mM. Theconstructed recombinant bacteria have important meanings for production of succinic acid.

Recombinant bacteria for producing succinic acid with glycerin being substrate and construction method and application thereof

Construction of an energy-conserving glycerol utilization pathways for improving anaerobic succinate production in Escherichia coli.

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Construction of a carbon-conserving pathway for glycolate production by synergetic utilization of acetate and glucose in Escherichia coli

New xylose transporters support the simultaneous consumption of glucose and xylose in Escherichia coli

Construction of Escherichia coli cell factories

Recombinant bacteria for producing succinic acid with glycerin being substrate and construction method and application thereof

Construction of an energy-conserving glycerol utilization pathways for improving anaerobic succinate production in Escherichia coli.