Next to plants, bacteria account for most of the biomass on Earth. They are found everywhere, although certain species thrive only in specific ecological niches. These microorganisms biosynthesize a plethora of both primary and secondary metabolites, defined, respectively, as those required for the growth and maintenance of cellular functions and those not required for survival but offering a selective advantage for the producer under certain conditions. As a result, bacterial fermentation has long been used to manufacture valuable natural products of nutritional, agrochemical and pharmaceutical interest. The interactions of secondary metabolites with their biological targets have been optimized by millions of years of evolution and they are, thus, considered to be privileged chemical structures, not only for drug discovery. During the last two decades, functional genomics has allowed for an in-depth understanding of the underlying biosynthetic logic of secondary metabolites. This has, in turn, paved the way for the unprecedented use of bacteria as programmable biochemical workhorses. In this Review, we discuss the multifaceted use of bacteria as biological factories in diverse applications and highlight recent advances in targeted genetic engineering of bacteria for the production of valuable bioactive compounds. Emphasis is on current advances to access nature’s abundance of natural products. This Review highlights bacteria as biological factories in diverse applications, with an emphasis on targeted genetic engineering for the production of bioactive natural products.

Bacteria as genetically programmable producers of bioactive natural products

Biobutanol offers several merits that make it as top-tier fuels to be discovered. Due to energy and environmental concern, traditional biobutanol production using Acetone-Butanol-Ethanol (ABE) fermentation have been revamped. An important facet for biobutanol production is selection of feedstock as a substrate. Currently, lignocellulosic biomass has attracted much interest globally due to its sustainable supply and non competitiveness with food. Cellulosic biobutanol production can be generated by using lignocellulosic residue such as agricultural crops and residue. Moreover, finding efficient pretreatment is another vital parameter in biobutanol production process. This review will briefly update the recent trends for lignocellulosic biomass as a substrate and feasible pretreatment that can be used for production of biobutanol.

Agro-industrial waste to biobutanol production: Eco-friendly biofuels for next generation

Clostridia are Gram-positive, spore-forming, obligate anaerobic bacteria that can produce solvents such as acetone, ethanol, and butanol, which can be used as biofuels or building block chemicals. Many successful attempts have been made to improve solvent yield and titer from sugars through metabolic engineering of solventogenic and acidogenic clostridia. More recently, cellulolytic and acetogenic clostridia have also attracted high interests for their ability to utilize low-cost renewable substrates such as cellulose and syngas. Process engineering such as in situ butanol recovery and consolidated bioprocessing (CBP) has been developed for improved solvent titer and productivity. This review focuses on metabolic and process engineering strategies for solvent production from sugars, lignocellulosic biomass, and syngas by various clostridia, including conventional solventogenic Clostridium acetobutylicum, engineered acidogens such as C. tyrobutyricum and C. cellulovorans, and carboxydotrophic acetogens such as C. carboxidivorans and C. ljungdahlii.

Engineering Clostridium for improved solvent production: recent progress and perspective

With the ever‐increasing demand for wearable electronics and energy‐saving technologies, self‐powered thermoelectric personal thermal management (PTM) has attracted extensive research interest. In this review, the unique characteristics of thermoelectric PTM comparing with other technologies are first highlighted, and the key parameters and fundamental functions of thermoelectric PTM are systematically summarized. Then, the advances in thermoelectric PTM are overviewed from the material design to the wearable device design viewpoints. Finally, the key challenges and future research directions of thermoelectric PTM, where both high‐performance flexible materials and proper device designs are in urgent need, are pointed out. This review will deliver a systematic understanding and guideline for thermoelectric PTM.

Multifunctional Wearable Thermoelectrics for Personal Thermal Management

Biofuel production from plant cell walls offers the potential for sustainable and economically attractive alternatives to petroleum-based products. In particular, Clostridium thermocellum is a promising host for consolidated bioprocessing (CBP) because of its strong native ability to ferment cellulose. We tested 12 different enzyme combinations to identify an n-butanol pathway with high titer and thermostability in C. thermocellum. The best producing strain contained the thiolase–hydroxybutyryl-CoA dehydrogenase–crotonase (Thl-Hbd-Crt) module from Thermoanaerobacter thermosaccharolyticum, the trans-enoyl-CoA reductase (Ter) enzyme from Spirochaeta thermophila and the butyraldehyde dehydrogenase and alcohol dehydrogenase (Bad-Bdh) module from Thermoanaerobacter sp. X514 and was able to produce 88 mg/L n-butanol. The key enzymes from this combination were further optimized by protein engineering. The Thl enzyme was engineered by introducing homologous mutations previously identified in Clostridium acetobutylicum. The Hbd and Ter enzymes were engineered for changes in cofactor specificity using the CSR-SALAD algorithm to guide the selection of mutations. The cofactor engineering of Hbd had the unexpected side effect of also increasing activity by 50-fold. Here we report engineering C. thermocellum to produce n-butanol. Our initial pathway designs resulted in low levels (88 mg/L) of n-butanol production. By engineering the protein sequence of key enzymes in the pathway, we increased the n-butanol titer by 2.2-fold. We further increased n-butanol production by adding ethanol to the growth media. By combining all these improvements, the engineered strain was able to produce 357 mg/L of n-butanol from cellulose within 120 h.

/pdf/metabolic-engineering-of-clostridium-thermocellum-for-n-4um1bt8e2g.pdf

Metabolic engineering of Clostridium thermocellum for n -butanol production from cellulose

Enhanced direct fermentation from food waste to butanol and hydrogen by an amylolytic Clostridium

The demand for cellulosic biofuels is on the rise because of the anticipation for sustainable energy and less greenhouse gas emissions in the future. However, production of cellulosic biofuels, especially cellulosic butanol, has been hampered by the lack of potent microbes that are capable of converting cellulosic biomass into biofuels. We report a wild-type Thermoanaerobacterium thermosaccharolyticum strain TG57, which is capable of using microcrystalline cellulose directly to produce butanol (1.93 g/liter) as the only final product (without any acetone or ethanol produced), comparable to that of engineered microbes thus far. Strain TG57 exhibits significant advances including unique genes responsible for a new butyrate synthesis pathway, no carbon catabolite repression, and the absence of genes responsible for acetone synthesis (which is observed as the main by-product in most Clostridium strains known today). Furthermore, the use of glucose analog 2-deoxyglucose posed a selection pressure to facilitate isolation of strain TG57 with deletion/silencing of carbon catabolite repressor genes-the ccr and xylR genes-and thus is able to simultaneously ferment glucose, xylose, and arabinose to produce butanol (7.33 g/liter) as the sole solvent. Combined analysis of genomic and transcriptomic data revealed unusual aspects of genome organization, numerous determinants for unique bioconversions, regulation of central metabolic pathways, and distinct transcriptomic profiles. This study provides a genome-level understanding of how cellulose is metabolized by T. thermosaccharolyticum and sheds light on the potential of competitive and sustainable biofuel production.

https://advances.sciencemag.org/content/advances/4/3/e1701475.full.pdf

Unique genetic cassettes in a Thermoanaerobacterium contribute to simultaneous conversion of cellulose and monosugars into butanol.

One of the main challenges of acetone–butanol–ethanol fermentation is to reduce acetone production with high butanol yield. Converting acetone into isopropanol is an alternative pathway to reduce fermentation by-products in the fermentation broth. Here, we aimed to cultivate a wild-type Clostridium strain with high isopropanol and butanol production and reveal its genome information. Clostridium beijerinckii strain BGS1 was found to be capable of producing 10.21 g/L butanol and 3.41 g/L isopropanol, higher than previously known wild-type isopropanol–butanol-producing Clostridium species. Moreover, culture BGS1 exhibited a broad carbon spectrum utilizing diverse sugars such as arabinose, xylose, galactose, cellobiose, and sucrose, with 9.61 g/L butanol and 2.57 g/L isopropanol generated from 60 g/L sucrose and less amount from other sugars. Based on genome analysis, protein-based sequence of strain BGS1 was closer to C. beijerinckii NCIMB 8052, reaching 90.82% similarity, while compared to C. beijerinckii DSM 6423, the similarity was 89.53%. In addition, a unique secondary alcohol dehydrogenase (sAdhE) was revealed in the genome of strain BGS1, which distinguished it from other Clostridium species. Average nucleotide identity analysis identified strain BGS1 belonging to C. beijerinckii. The transcription profile and enzymatic activity of sAdhE proved its function of converting acetone into isopropanol. Clostridium beijerinckii strain BGS1 is a potential candidate for industrial isopropanol and butanol production. Its genome provides unique information for genetic engineering of isopropanol–butanol-producing microorganisms.

/pdf/characterization-and-genome-analysis-of-a-butanol-355s6plvmf.pdf

Chen Zhang

Papers

Enhanced direct fermentation from food waste to butanol and hydrogen by an amylolytic Clostridium

Unique genetic cassettes in a Thermoanaerobacterium contribute to simultaneous conversion of cellulose and monosugars into butanol.

Characterization and genome analysis of a butanol-isopropanol-producing Clostridium beijerinckii strain BGS1.

All-Soft and Stretchable Thermogalvanic Gel Fabric for Antideformity Body Heat Harvesting Wearable

A Fast Autonomous Healing Magnetic Elastomer for Instantly Recoverable, Modularly Programmable, and Thermorecyclable Soft Robots