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Engineering Sphingobium sp. to Accumulate Various Carotenoids Using Agro-Industrial Byproducts.

ABSTRACT: Carotenoids represent the most abundant lipid-soluble phytochemicals that have been shown to exhibit benefits for nutrition and health. The production of natural carotenoids is not yet cost effective to compete with chemically synthetic ones. Therefore, the demand for natural carotenoids and improved efficiency of carotenoid biosynthesis has driven the investigation of metabolic engineering of native carotenoid producers. In this study, a new Sphingobium sp. was isolated, and it was found that it could use a variety of agro-industrial byproducts like soybean meal, okara, and corn steep liquor to accumulate large amounts of nostoxanthin. Then we tailored it into three mutated strains that instead specifically accumulated ∼5 mg/g of CDW of phytoene, lycopene, and zeaxanthin due to the loss-of-function of the specific enzyme. A high-efficiency targeted engineering carotenoid synthesis platform was constructed in Escherichia coli for identifying the functional roles of candidate genes of carotenoid biosynthetic pathway in Sphingobium sp. To further prolong the metabolic pathway, we engineered the Sphingobium sp. to produce high-titer astaxanthin (10 mg/g of DCW) through balance in the key enzymes β-carotene ketolase (BKT) and β-carotene hydroxylase (CHY). Our study provided more biosynthesis components for bioengineering of carotenoids and highlights the potential of the industrially important bacterium for production of various natural carotenoids.

SUBMITTER: Liu M

PROVIDER: S-EPMC8600064 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Humic-like substances (HLSs) isolated by alkaline oxidative hydrolysis from lignin-rich agro-industrial residues have been shown to exert biostimulant activity toward maize (Zea mays L.) germination and early growth. The definition of a quantitative structure-activity relationship (QSAR) between HLS and their bioactivity could be useful to predict their biological properties and tailor plant biostimulants for specific agronomic and industrial uses. Here, we created several projection on latent structure (PLS) regression by using published analytical data on the molecular composition of lignin-derived HLS obtained by both 13C-CPMAS-NMR spectra directly on samples and 31P-NMR spectra after derivatization of hydroxyl functions with a P-containing reagent (2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane). These spectral data were used to model the effect of HLS on the elongation of primary root, lateral seminal roots, total root apparatus, and coleoptile of maize. The 13C-CPMAS-NMR data suggested that methoxyl and aromatic moieties positively affected plant growth, while the carboxyl/esterified functions showed a negative impact on the overall seedling development. Alkyl C seems to promote Col elongation while concomitantly reducing that of the root system. Additionally, 31P-NMR-derived spectra revealed that the elongation of roots and Col were enhanced by the occurrence of aliphatic hydroxyl groups, and guaiacyl and p-Hydroxyphenyl lignin monomers. The PLS models based on raw dataset from 13C-CPMAS-NMR spectra explained more than 74% of the variance for the length of lateral seminal roots, total root system and coleoptile, while other parameters derived from 13C-CPMAS-NMR spectra, namely the Hydrophobicity and Hydrophilicity of materials were necessary to explain 83% of the variance of the primary root length. The results from 31P-NMR spectra explained the observed biological variance by 90, 96, 96, and 93% for the length of primary root, lateral seminal roots, total root system and coleoptile, respectively. This work shows that different NMR spectroscopy techniques can be used to build up PLS models which can predict the bioactivity of lignin-derived HLS toward early growth of maize plants. The established QSAR may also be exploited to enhance by chemical techniques the bioactive properties of HLS and enhance their plant stimulation capacity.

Project description:Background2,3-Butanediol (BD) is a largely used fossil-based platform chemical. The yield and productivity of bio-based BD fermentative production must be increased and cheaper substrates need to be identified, to make bio-based BD production more competitive. As BD bioproduction occurs under microaerobic conditions, a fine tuning and control of the oxygen transfer rate (OTR) is crucial to maximize BD yield and productivity. Very few studies on BD bioproduction focused on the use of non-pathogenic microorganisms and of byproducts as substrate. The goal of this work was to optimize BD bioproduction by the non-pathogenic strain Bacillus licheniformis ATCC9789 by (i) identifying the ranges of volumetric and biomass-specific OTR that maximize BD yield and productivity using standard sugar and protein sources, and (ii) performing a preliminary evaluation of the variation in process performances and cost resulting from the replacement of glucose with molasses, and beef extract/peptone with chicken meat and bone meal, a byproduct of the meat production industry.ResultsOTR optimization with an expensive, standard medium containing glucose, beef extract and peptone revealed that OTRs in the 7-15 mmol/L/h range lead to an optimal BD yield (0.43 ± 0.03 g/g) and productivity (0.91 ± 0.05 g/L/h). The corresponding optimal range of biomass-specific OTR was equal to 1.4-7.9 [Formula: see text], whereas the respiratory quotient ranged from 1.8 to 2.5. The switch to an agro-industrial byproduct-based medium containing chicken meat and bone meal and molasses led to a 50% decrease in both BD yield and productivity. A preliminary economic analysis indicated that the use of the byproduct-based medium can reduce by about 45% the BD production cost.ConclusionsA procedure for OTR optimization was developed and implemented, leading to the identification of a range of biomass-specific OTR and respiratory quotient to be used for the scale-up and control of BD bioproduction by Bacillus licheniformis. The switch to a byproduct-based medium led to a relevant decrease in BD production cost. Further research is needed to optimize the process of BD bioproduction from the tested byproduct-based medium.

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Engineering Sphingobium sp. to Accumulate Various Carotenoids Using Agro-Industrial Byproducts.

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