| 1. |
- Abdelaziz, Omar Y., et al.
(författare)
-
Biological valorization of low molecular weight lignin
- 2016
-
Ingår i: Biotechnology Advances. - : Elsevier BV. - 0734-9750. ; 34:8, s. 1318-1346
-
Forskningsöversikt (refereegranskat)abstract
- Lignin is a major component of lignocellulosic biomass and as such, it is processed in enormous amounts in the pulp and paper industry worldwide. In such industry it mainly serves the purpose of a fuel to provide process steam and electricity, and to a minor extent to provide low grade heat for external purposes. Also from other biorefinery concepts, including 2nd generation ethanol, increasing amounts of lignin will be generated. Other uses for lignin – apart from fuel production – are of increasing interest not least in these new biorefinery concepts. These new uses can broadly be divided into application of the polymer as such, native or modified, or the use of lignin as a feedstock for the production of chemicals. The present review focuses on the latter and in particular the advances in the biological routes for chemicals production from lignin. Such a biological route will likely involve an initial depolymerization, which is followed by biological conversion of the obtained smaller lignin fragments. The conversion can be either a short catalytic conversion into desired chemicals, or a longer metabolic conversion. In this review, we give a brief summary of sources of lignin, methods of depolymerization, biological pathways for conversion of the lignin monomers and the analytical tools necessary for characterizing and evaluating key lignin attributes.
|
|
| 2. |
- Abdelaziz, Omar Y., et al.
(författare)
-
Membrane filtration of alkali-depolymerised kraft lignin for biological conversion
- 2019
-
Ingår i: Bioresource Technology Reports. - : Elsevier BV. - 2589-014X. ; 7
-
Tidskriftsartikel (refereegranskat)abstract
- In this study, we have investigated the possibility of membrane filtration as a means for obtaining a fraction containing mainly low-molecular-weight (LMW) compounds from depolymerised lignin (DL) for subsequent microbial conversion. A DL stream from continuous-mode alkali depolymerisation of a softwood kraft lignin produced at a temperature of 220 °C and a residence time of 2 min, using a NaOH/lignin weight ratio of 1 with 5 wt% lignin loading was fractionated using a polymeric membrane with a molecular weight cut-off of 500–700 Da. The permeate (DLP) volume recovery of LMW phenolics (250–450 Da) was 70% after filtration for 3.7 h. The DLP was used as a carbon source for growth of three bacterial strains; Pseudomonas fluorescens, P. putida EM42 and Rhodococcus opacus, and good growth was obtained by the first two microorganisms. This proof-of-concept study demonstrates a novel strategy for technical lignin valorisation by combining depolymerisation, nanofiltration and bioconversion.
|
|
| 3. |
- Abdelaziz, Omar Y., et al.
(författare)
-
Oxidative Depolymerization of Kraft Lignin for Microbial Conversion
- 2019
-
Ingår i: ACS Sustainable Chemistry and Engineering. - : American Chemical Society (ACS). - 2168-0485. ; 7:13, s. 11640-11652
-
Tidskriftsartikel (refereegranskat)abstract
- The valorization of lignin is being increasingly recognized as crucial to improve the economic viability of integrated biorefineries. Because of its inherent heterogeneity and recalcitrance, lignin has been treated as a waste product in the pulp and paper industry, but new technologies are now being explored to transform lignin into a sustainable resource and enhance its value chain. In the present study, alkaline oxidative depolymerization was investigated as a potential form of pretreatment to enable further biological conversion of LignoBoost kraft lignin (LB). LB lignin oxidation reactions were studied at various temperatures (120-200 °C) and O2 partial pressures (3-15 bar) to identify the optimal conditions for obtaining a biocompatible, oxidatively depolymerized lignin (ODLB) stream. The low molecular weight compounds resulting from this treatment consisted mainly of aromatic monomers and carboxylic acids. The highest yield of aromatic monomers, 3 wt %, was obtained at 160 °C and 3 bar O2. The yield of carboxylic acids increased with both increasing temperature and O2 pressure, exceeding 13% under the harshest conditions investigated. The growth of four aromatic-catabolizing bacterial strains was examined on reaction product mixtures, all of which showed growth on agar plates utilizing ODLB as the sole source of carbon and energy. Rhodococcus opacus and Sphingobium sp. SYK-6 were found to consume most of the aromatic monomers present in the ODLB (e.g., vanillin, vanillate, acetovanillone, and guaiacol). The findings of this study indicate that pretreatment by oxidative depolymerization has potential in the biological valorization of technical lignin streams, for the production of valuable chemicals and materials.
|
|
| 4. |
- Brink, Daniel P., et al.
(författare)
-
Mapping the diversity of microbial lignin catabolism : experiences from the eLignin database
- 2019
-
Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 0175-7598 .- 1432-0614. ; , s. 3979-4002
-
Forskningsöversikt (refereegranskat)abstract
- Lignin is a heterogeneous aromatic biopolymer and a major constituent of lignocellulosic biomass, such as wood and agricultural residues. Despite the high amount of aromatic carbon present, the severe recalcitrance of the lignin macromolecule makes it difficult to convert into value-added products. In nature, lignin and lignin-derived aromatic compounds are catabolized by a consortia of microbes specialized at breaking down the natural lignin and its constituents. In an attempt to bridge the gap between the fundamental knowledge on microbial lignin catabolism, and the recently emerging field of applied biotechnology for lignin biovalorization, we have developed the eLignin Microbial Database (www.elignindatabase.com), an openly available database that indexes data from the lignin bibliome, such as microorganisms, aromatic substrates, and metabolic pathways. In the present contribution, we introduce the eLignin database, use its dataset to map the reported ecological and biochemical diversity of the lignin microbial niches, and discuss the findings.
|
|
| 5. |
- García-Hidalgo, Javier, et al.
(författare)
-
Identification of the two-component guaiacol demethylase system from Rhodococcus rhodochrous and expression in Pseudomonas putida EM42 for guaiacol assimilation
- 2019
-
Ingår i: AMB Express. - : Springer Science and Business Media LLC. - 2191-0855. ; 9:1
-
Tidskriftsartikel (refereegranskat)abstract
- A diversity of softwood lignin depolymerization processes yield guaiacol as the main low molecular weight product. This key aromatic compound can be utilized as a carbon source by several microbial species, most of which are Gram positive bacteria. Microbial degradation of guaiacol is known to proceed initially via demethylation to catechol, and this reaction is catalyzed by cytochrome P450 monooxygenases. These enzymes typically require a set of redox partner proteins, whose number and identities were not described until very recently in the case of guaiacol. In this work we identified two proteins involved in guaiacol demethylation by the actinomycete Rhodococcus rhodochrous. Additionally, we constructed four different polycistronic operons carrying combinations of putative redox partners of this guaiacol demethylation system in an inducible expression plasmid that was introduced into the Gram negative host Pseudomonas putida EM42, and the guaiacol consumption dynamics of each resulting strain were analyzed. All the polycistronic operons, expressing a cytochrome P450 together with a putative ferredoxin reductase from R. rhodochrous and putative ferredoxins from R. rhodochrous or Amycolatopsis ATCC 39116 enabled P. putida EM42 to metabolize and grow on guaiacol as the sole carbon source.
|
|
| 6. |
- García-Hidalgo, Javier, et al.
(författare)
-
Vanillin Production in pseudomonas : Whole-genome sequencing of pseudomonas sp. strain 9.1 and reannotation of pseudomonas putida CalA as a vanillin reductase
- 2020
-
Ingår i: Applied and Environmental Microbiology. - 0099-2240. ; 86:6
-
Tidskriftsartikel (refereegranskat)abstract
- Microbial degradation of lignin and its related aromatic compounds has great potential for the sustainable production of chemicals and bioremediation of contaminated soils. We previously isolated Pseudomonas sp. strain 9.1 from historical waste deposits (forming so-called fiber banks) released from pulp and paper mills along the Baltic Sea coast. The strain accumulated vanillyl alcohol during growth on vanillin, and while reported in other microbes, this phenotype is less common in wild-type pseudomonads. As the reduction of vanillin to vanil- lyl alcohol is an undesired trait in Pseudomonas strains engineered to accumulate vanillin, connecting the strain 9.1 phenotype with a genotype would increase the fundamental understanding and genetic engineering potential of microbial vanillin metabolism. The genome of Pseudomonas sp. 9.1 was sequenced and assembled. Annotation identified oxidoreductases with homology to Saccharomyces cerevisiae alcohol dehydrogenase ScADH6p, known to reduce vanillin to vanillyl alcohol, in both the 9.1 genome and the model strain Pseudomonas putida KT2440. Recombinant expression of the Pseudomonas sp. 9.1 FEZ21_09870 and P. putida KT2440 PP_2426 (calA) genes in Escherichia coli revealed that these open reading frames encode aldehyde reductases that convert vanillin to vanillyl alcohol, and that P. putida KT2440 PP_3839 encodes a coniferyl alcohol dehydrogenase that oxidizes coniferyl alcohol to coniferyl aldehyde (i.e., the function previ-ously assigned to calA). The deletion of PP_2426 in P. putida GN442 engineered to accumulate vanillin resulted in a decrease in by-product (vanillyl alcohol) yield from 17% to -1%. Based on these results, we propose the reannotation of PP_2426 and FEZ21_09870 as areA and PP_3839 as calA-II. IMPORTANCE Valorization of lignocellulose (nonedible plant matter) is of key interest for the sustainable production of chemicals from renewable resources. Lignin, one of the main constituents of lignocellulose, is a heterogeneous aromatic biopolymer that can be chemically depolymerized into a heterogeneous mixture of aromatic building blocks; those can be further converted by certain microbes into value-added aromatic chemicals, e.g., the flavoring agent vanillin. We previously isolated a Pseudomonas sp. strain with the (for the genus) unusual trait of vanil- lyl alcohol production during growth on vanillin. Whole-genome sequencing of the isolate led to the identification of a vanillin reductase candidate gene whose deletion in a recombinant vanillin-accumulating P. putida strain almost completely alleviated the undesired vanillyl alcohol by-product yield. These results represent an important step toward biotechnological production of vanillin from lignin using bacterial cell factories.
|
|
| 7. |
- Ravi, Krithika, et al.
(författare)
-
Bacterial conversion of depolymerized Kraft lignin
- 2019
-
Ingår i: Biotechnology for Biofuels. - : Springer Science and Business Media LLC. - 1754-6834. ; 12
-
Tidskriftsartikel (refereegranskat)abstract
- BackgroundLignin is a potential feedstock for microbial conversion into various chemicals. However, the microbial degradation rate of native or technical lignin is low, and chemical depolymerization is needed to obtain reasonable conversion rates. In the current study, nine bacterial strains belonging to the Pseudomonas and Rhodococcus genera were evaluated for their ability to grow on alkaline-treated softwood lignin as a sole carbon source.ResultsPseudomonas fluorescens DSM 50090 and Rhodococcus opacus DSM1069 showed the best growth of the tested species on plates with lignin. Further evaluation of P. fluorescens and R. opacus was made in liquid cultivations with depolymerized softwood Kraft lignin (DL) at a concentration of 1 g/L. Size-exclusion chromatography (SEC) showed that R. opacus consumed most of the available lower-molecular weight compounds (approximately 0.1–0.4 kDa) in the DL, but the weight distribution of larger fractions was almost unaffected. Importantly, the consumed compounds included guaiacol—one of the main monomers in the DL. SEC analysis of P. fluorescens culture broth, in contrast, did not show a large conversion of low-molecular weight compounds, and guaiacol remained unconsumed. However, a significant shift in molecular weight distribution towards lower average weights was seen after cultivation with P. fluorescens.ConclusionsRhodococcus opacus and P. fluorescens were identified as two potential microbial candidates for the conversion/consumption of base-catalyzed depolymerized lignin, acting on low- and high-molecular weight lignin fragments, respectively. These findings will be of relevance for designing bioconversion of softwood Kraft lignin.
|
|
| 8. |
- Ravi, Krithika, et al.
(författare)
-
Biological conversion of aromatic monolignol compounds by a Pseudomonas isolate from sediments of the Baltic Sea
- 2018
-
Ingår i: AMB Express. - : Springer Science and Business Media LLC. - 2191-0855. ; 8:1
-
Tidskriftsartikel (refereegranskat)abstract
- Bacterial strains were isolated from the sediments of the Baltic Sea using ferulic acid, guaiacol or a lignin-rich softwood waste stream as substrate. In total nine isolates were obtained, five on ferulic acid, two on guaiacol and two on a lignin-rich softwood stream as a carbon source. Three of the isolates were found to be Pseudomonas sp. based on 16S rRNA sequencing. Among them, isolate 9.1, which showed the fastest growth in defined M9 medium, was tentatively identified as a Pseudomonas deceptionensis strain based on the gyrB sequencing. The growth of isolate 9.1 was further examined on six selected lignin model compounds (ferulate, p-coumarate, benzoate, syringate, vanillin and guaiacol) from different upper funneling aromatic pathways and was found able to grow on four out of these six compounds. No growth was detected on syringate and guaiacol. The highest specific growth and uptake rates were observed for benzoate (0.3 h−1 and 4.2 mmol gCDW −1 h−1) whereas the lowest were for the compounds from the coniferyl branch. Interestingly, several pathway intermediates were excreted during batch growth. Vanillyl alcohol was found to be excreted during growth on vanillin. Several other intermediates like cis,cis-muconate, catechol, vanillate and 4-hydroxybenzoate from the known bacterial catabolic pathways were excreted during growth on the model compounds.
|
|
| 9. |
- Ravi, Krithika, et al.
(författare)
-
Conversion of lignin model compounds by Pseudomonas putida KT2440 and isolates from compost
- 2017
-
Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 1432-0614 .- 0175-7598. ; 101:12, s. 5059-5070
-
Tidskriftsartikel (refereegranskat)abstract
- Starting from mature vegetable compost, four bacterial strains were selected using a lignin-rich medium. 16S ribosomal RNA identification of the isolates showed high score similarity with Pseudomonas spp. for three out of four isolates. Further characterization of growth on mixtures of six selected lignin model compounds (vanillin, vanillate, 4-hydroxybenzoate, p-coumarate, benzoate, and ferulate) was carried out with three of the Pseudomonas isolates and in addition with the strain Pseudomonas putida KT2440 from a culture collection. The specific growth rates on benzoate, p-coumarate, and 4-hydroxybenzoate were considerably higher (0.26–0.27 h−1) than those on ferulate and vanillate (0.21 and 0.22 h−1), as were the uptake rates.There was no direct growth of P. putida KT2440 on vanillin, but instead, vanillin was rapidly converted in to vanillate at a rate of 4.87 mmol (gCDW h)−1 after which the accumulated vanillate was taken up. The growth curve reflected a diauxic growthwhen mixtures of the model compounds were used as carbon source. Vanillin, 4-hydroxybenzoate, and benzoate were preferentially consumed first, whereas ferulate was always the last substrate to be taken in. These results contribute to a better understanding of the aromatic metabolism of P. putida in terms of growth and uptake rates, which will be helpful for the utilization of these bacteria as cell factories for upgrading lignin-derived mixtures of aromatic molecules.
|
|
| 10. |
- Ravi, Krithika, et al.
(författare)
-
Physiological characterization and sequence analysis of a syringate-consuming Actinobacterium
- 2019
-
Ingår i: Bioresource Technology. - : Elsevier BV. - 0960-8524. ; 285
-
Tidskriftsartikel (refereegranskat)abstract
- Hardwood lignin is made of up to 75% syringyl-units and the bioconversion of syringate and syringaldehyde is therefore of considerable interest for biological valorization of lignin. In the current study, we have isolated a syringate-consuming bacterium identified as Microbacterium sp. RG1 and characterized its growth on several lignin model compounds. Growth was observed on syringate, 3-O-methylgallate, vanillate, 4-hydroxybenzoate, ferulate and p-coumarate. Toxic aromatic aldehydes such as vanillin and syringaldehyde were converted to their respective alcohols/acids which were eventually consumed with a maximum specific uptake rate of 0.02 and 0.1 mmol (g CDW h) −1 respectively. The isolate was further subjected to whole genome sequencing and putative genes related to the metabolism of syringyl-compounds were mapped for the first time in a Gram-positive bacterium. These findings will be of high significance when designing future host microorganisms and bioprocesses for the efficient valorization of pre-treated lignin feedstocks.
|
|
