Utilization of a Wheat Sidestream for 5-Aminovalerate Production in Corynebacterium glutamicum

Abstract

Production of plastics from petroleum-based raw materials extensively contributes to global pollution and CO2 emissions. Biotechnological production of functionalized monomers can reduce the environmental impact, in particular when using industrial sidestreams as feedstocks. Corynebacterium glutamicum, which is used in the million-ton-scale amino acid production, has been engineered for sustainable production of polyamide monomers. In this study, wheat sidestream concentrate (WSC) from industrial starch production was utilized for production of l-lysine–derived bifunctional monomers using metabolically engineered C. glutamicum strains. Growth of C. glutamicum on WSC was observed and could be improved by hydrolysis of WSC. By heterologous expression of the genes xylAXcBCg (xylA from Xanthomonas campestris) and araBADEc from E. coli, xylose, and arabinose in WSC hydrolysate (WSCH), in addition to glucose, could be consumed, and production of l-lysine could be increased. WSCH-based production of cadaverine and 5-aminovalerate (5AVA) was enabled. To this end, the lysine decarboxylase gene ldcCEc from E. coli was expressed alone or for conversion to 5AVA cascaded either with putrescine transaminase and dehydrogenase genes patDAEc from E. coli or with putrescine oxidase gene puoRq from Rhodococcus qingshengii and patDEc. Deletion of the l-glutamate dehydrogenase–encoding gene gdh reduced formation of l-glutamate as a side product for strains with either of the cascades. Since the former cascade (ldcCEc-patDAEc) yields l-glutamate, 5AVA production is coupled to growth by flux enforcement resulting in the highest 5AVA titer obtained with WSCH-based media.

Rational Engineering of Non-Ubiquinone Containing Corynebacterium glutamicum for Enhanced Coenzyme Q10 Production

“…After CoQ 10 production was successfully improved, the strain UBI6-Rs was cultivated with a hydrolysate of the alternative feedstock wheat side stream concentrate (WSCH) [ 40 ]. In a sustainable circular economy, side streams of industrial production processes can provide excellent alternative feedstocks for microbial production containing macro- and micronutrients [ 41 ].…”

“…With respect to alternative feedstocks that are not competitive with food or feed, C. glutamicum has been employed and engineered for many different substrates. Here, we demonstrated the successful production of CoQ 10 from a wheat side stream-based hydrolysate that has been utilized previously for the production of 5-aminovalerate [ 40 ] and l -2-hydroxyglutarate [ 22 ]. As well as the wheat side stream, access to numerous monomeric and polymeric carbon sources has been established, e.g., xylose, arabinose, mannose, starch, lignocellulose, N -acetylglucosamine, and alginate, which can be derived from hydrolysates of sustainable second generation feedstocks such as spent sulfite liquor, Miscanthus biomass, brown seaweed, corn straw, rice straw, and shrimp waste [ 63 , 64 , 65 , 66 ].…”

“…The minimal medium was supplemented with 40 g L −1 glucose as sole carbon source, 1 mM IPTG to induce gene expression of ddsA from pRG_Duet2 [ 69 ] and all genes from pEC-XT99A [ 70 ] and pEKEx3 [ 71 ], 0.25 µg mL −1 anhydrotetracycline (ATc) to induce gene expression of ubiA from pRG_Duet2, and respective antibiotics. For cultivation in WSCH medium, 80% ( v / v ) hydrolysate (from 190 g L −1 WSC, [ 40 ]) was supplemented with inducers and antibiotics as described above, 20 g L −1 ammonium sulfate and 42 g L −1 MOPS to a final glucose concentration of 33.2 g L −1 . OD 600 was measured using a V-1200 spectrophotometer (VWR, Radnor, PA, United States).…”

Parishin From Gastrodia Elata Ameliorates Aging Phenotype in Mice in a Gut Microbiota-Related Manner

“…These findings are consistent with the results of the fecal metabolome analysis in which parishin treatment reversed the aging-induced depletion of 3-(3-hydroxyphenyl)-propanoic acid, L-proline, 4-hydroxybenzeneacetic acid and 5-aminovaleric acid. 5-Aminovaleric acid, the γ-aminobutyric acid (GABA) analog, is an important intermediate in the pathways of L-lysine degradation and L-proline degradation ( Burgardt et al, 2021 ; Cheng et al, 2021 ); the microbial flavonoid metabolite 4-hydroxybenzeneacetic acid, which possesses anxiolytic effects, is generated from gut microbial polyphenol and amino acid metabolism, and is an important intermediate in the pathways of L-tyrosine degradation and automatic biogenic amine degradation ( Zabela et al, 2016 ; Orabi et al, 2021 ); 3-(3-hydroxyphenyl)-propanoic acid, which has been reported to exhibit hepatoprotective activities by regulating oxidative stress, lipid peroxidation, and inflammatory responses ( Cui et al, 2018 ), is an intermediate in the phosphatidylcholine resynthesis pathway and in the mitochondrial NADPH production pathway of the cofactor, carrier, and vitamin biosynthesis superfamily. In addition, the parishin treatment-improved metabolite β-sitosterol is a plant-derived natural bioactive compound with various pharmacological activities, such as hypolipidemic, anti-inflammatory, antitumor, tissue repair and antiaging activities ( Haiyuan et al, 2019 ; Cheng et al, 2020 ), and is used to treat fatty liver, type II diabetes, and arthritis ( Gumede et al, 2020 ; Ramalingam et al, 2020 ).…”

Metabolic Engineering for Valorization of Agri- and Aqua-Culture Sidestreams for Production of Nitrogenous Compounds by Corynebacterium glutamicum

“…glutamicum consortium has been engineered for growth and production with starch ( Sgobba et al, 2018b ). The remaining challenges are that starchy biomass does have competing uses as food and feed, that debranching enzymes such as pullulanase have to be secreted or that starch has to be hydrolyzed to access cyclodextrin and branched starch portions ( Burgardt et al, 2021 ).…”