Biohydrogen production, storage, and delivery: A comprehensive overview of current strategies and limitations

A review on biological biohydrogen production: Outlook on genetic strain enhancements, reactor model and techno-economics analysis.

Recent advances in process improvement of dark fermentative hydrogen production through metabolic engineering strategies

Biomass waste must be treated and disposed off properly, as it concerns the possibility of deposition in the environment. This could worsen the ecosystem by promoting the growth of pathogens, producing poisonous fumes, releasing ammonia and other dangerous gases into the atmosphere. Therefore, researchers are exploring different direct biomass waste applications. One such application is biotechnological options with financial and environmental benefits like lowering the consumption of fossil fuels, reducing harmful emissions, accelerating the synthesis of low-cost raw materials, and fostering an environment that is favourable for a broad range of microorganisms. In recent years, biofuel has emerged as the primary source of bio-energy by replacing fossil fuels that can meet the global energy demands and lower greenhouse gas (GHG) emissions. This review explores the potential of biomass waste conversion techniques and examines the production of biofuels to achieve zero carbon emissions. Critical research has also been done to improve the anaerobic treatment method of the biochemical conversion method. Apart from these, the advancements in the study of metabolic engineering techniques and artificial intelligence in the production of biofuel and strategies along with microbial chassis to enhance production of biofuel in an efficient manner were also studied. Finally, challenges and future perspectives of biofuels were delineated. 

A review on emerging technologies and machine learning approaches for sustainable production of biofuel from biomass waste

Pyruvate is a hub of various endogenous metabolic pathways, including glycolysis, TCA cycle, amino acid, and fatty acid biosynthesis. It has also been used as a precursor for pyruvate-derived compounds such as acetoin, 2,3-butanediol (2,3-BD), butanol, butyrate, and L-alanine biosynthesis. Pyruvate and derivatives are widely utilized in food, pharmaceuticals, pesticides, feed additives, and bioenergy industries. However, compounds such as pyruvate, acetoin, and butanol are often chemically synthesized from fossil feedstocks, resulting in declining fossil fuels and increasing environmental pollution. Metabolic engineering is a powerful tool for producing eco-friendly chemicals from renewable biomass resources through microbial fermentation. Here, we review and systematically summarize recent advances in the biosynthesis pathways, regulatory mechanisms, and metabolic engineering strategies for pyruvate and derivatives. Furthermore, the establishment of sustainable industrial synthesis platforms based on alternative substrates and new tools to produce these compounds is elaborated. Finally, we discuss the potential difficulties in the current metabolic engineering of pyruvate and derivatives and promising strategies for constructing efficient producers.

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Metabolic Engineering of Microorganisms to Produce Pyruvate and Derived Compounds

Regulation of the NADH supply by nuoE deletion and pncB overexpression to enhance hydrogen production in Enterobacter aerogenes

As a valuable platform chemical, 2,3-Butanediol (2,3-BDO) has a variety of industrial applications, and its microbial production is particularly attractive as an alternative to petroleum-based production. In this study, the regulation of intracellular carbon flux and NADH/NAD+ was used to increase the 2,3-BDO production of Enterobacter aerogenes. The genes encoding lactate dehydrogenase (ldh) and pyruvate formate lyase (pfl) were disrupted using the λ-Red recombination method and CRISPR-Cas9 to reduce the production of several byproducts and the consumption of NADH. Knockout of ldh or pfl increased intracellular NADH/NAD+ by 111 % and 113 %, respectively. Moreover, two important genes in the 2,3-BDO biosynthesis pathway, acetolactate synthase (budB) and acetoin reductase (budC), were overexpressed in E. aerogenes to further amply the metabolic flux toward 2,3-BDO production. And the overexpression of budB or budC increased intracellular NADH/NAD+ by 46 % and 57 %, respectively. In shake-flask cultivation with sucrose as carbon source, the 2,3-BDO titer of the IAM1183-LPBC was 3.55 times that of the wild type. In the 5-L fermenter, the maximal 2,3-BDO production produced by the IAM1183-LPBC was 2.88 times that of the original strain. This work offers new ideas for promoting the biosynthesis of 2,3-BDO for industrial applications. 

Regulation of carbon flux and NADH/NAD+ supply to enhance 2,3-butanediol production in Enterobacter aerogenes.

Metabolic regulation of NADH supply and hydrogen production in Enterobacter aerogenes by multi-gene engineering

Dark fermentation is considered as one of the most practical biological hydrogen production methods. However, current productivity and yield are still not economically viable for industrial applications. This biological process must be improved through multiple strategies, of which screening for more effective microbial strains is an important aspect. Here, based on the hydrogen production pathway of E. aerogenes, we describe three strategies to improve hydrogen production by effectively regulating the anaerobic metabolism of E. aerogenes through genetic modification. This protocol describes in detail how to obtain NADH dehydrogenase-damaged mutants and overexpress Nad synthase genes using the CRISPR-Cas9 gene editing system. In addition, the overexpression of small RNA RyhB was achieved and verified by Northern Blot. This protocol is of great significance for the study of genetic engineering operation in E. aerogenes and other bacteria, and also provides theoretical guidance and technical support for the study of E. aerogenes biological hydrogen production. 

Integrated strategy of CRISPR-Cas9 gene editing and small RNA RhyB regulation in Enterobacter aerogenes: A novel protocol for improving biohydrogen production

Characterization and bioactive component analysis of filamentous bacterium XJ-16

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